Association between apolipoprotein E polymorphism and lipid metabolism in patients with cerebral infarction

AIM:To study the relationship between ApoE polymorphism and lipid metabolism of patients with cerebral infarction. METHODS:ApoE phenotype was determined in 110 patients with cerebral infarction and 60 normal controls by isoelectric focusing and immunoblotting. The TC, TG and HDL-C levels in serum of these subjects were measured with enzymes methods, ApoA I and ApoB levels with rocket immunoelectrophoresis methods, ApoE and Lp(a) levels with ELISA methods. RESULTS:The differences of the ApoE polymorphism distribution and ApoE allele frequencies (P＜0.05) occurred between two groups. The frequence of ApoE ε4 allele in patients with cerebral infarction was significantly higher than those in the normal controls (P＜0.05). However, ApoE 3/3 phenotypes was significantly lower (P＜0.05). Comparison of values of serum lipid with various ApoE phenotyoe among patients with cerebral infarction revealed that there was correlation between ApoE polymorphism and TC(P＜0.05), TG(P＜0.05), HDL-C(P＜0.05), LDL-C(P＜0.05), ApoA I(P＜0.05), ApoB(P＜0.05), ApoE(P＜0.05)and Lp(a)(P＜0.05). Patients carrying ε4 were associated with increased TC, LDL-C, ApoB and Lp(a), while those with ε2 were assiociated with decreased TC, LDL-C, and ApoB. Patients carrying ε2 were associated with increased TG, HDL-C, ApoAⅠ, and ApoE.CONCLUSIONS:ApoEε4 allele was associated with the development of cerebral infarction. ApoE polyphorphism affects lipid metabolism of cerebral infarction patients.     

Distribution characteristics of polymorphisms of interleukin-22 gene in Guangxi population and their association with serum lipid levels

AIM: To investigate the distribution characteristics of interleukin-22 (IL-22) gene rs2227485C/T and rs2227491A/G polymorphisms in Guangxi people and the distribution differences with other ethnic groups, and to explore the difference levels of common lipid indexes in different genotypes. METHODS: SNaPshot technique and DNA sequencing were used in 280 Guangxi persons to examine IL-22 genotypes and to analyzed the distribution frequencies of allele and genotype in these sites. The distribution frequencies in different sexes, and the differences between groups and diffe-rence levels of common lipid indexes in different genotypes were analyzed statistically. RESULTS: Three genotypes of CC, CT and TT were found in rs2227485C/T with the frequency distribution of 17.1%, 49.3% and 33.6%, respectively. No significant difference between different sexes of each genotype and allele frequency in the Guangxi population was observed (P>0.05). Compared with the distribution frequencies of genotype and allele in HapMap-TSI, HapMap-HCB, HapMap-JPT and HapMap-MEX, those in Guangxi population showed statistically significant differences (P<0.05). Three genotypes of AA, AG and GG were found in rs2227491A/G with the frequency distribution of 16.1%, 52.8% and 31.1%, respectively. There was no significant difference between different sexes of each genotype and allele frequency in the Guangxi population (P>0.05). The significant differences of genotype frequencies among Guangxi population, HapMap-TSI, HapMap-JPT and HapMap-MEX were detected (P<0.05). Compared with the other 4 populations, allele frequencies in Guangxi population had significant difference (P <0.05). There were significant differences in the levels of HDL-C and LDL-C among the 3 genotypes of rs2227491A/G. The level of HDL-C had difference between AG/AA genotype and GG genotype. In addition, the level of LDL-C had difference between AG/GG genotype and AA genotype (P<0.05). CONCLUSION: rs2227485C/T and rs2227491A/G polymorphisms of IL-22 gene have differences in different populations. The rs2227491A/G polymorphism may be associated with serum lipid levels.     

Effects of a pentanucleotide repeat polymorphism of the apolipoprotein (a)gene on serum levels of lipoproteins in healthy Han population

AIM:To understand the relationship between pentanucleotide repeat(PNR) polymorphism of the apolipoprotein(a) gene and lipoprotein natures of normal Han population. METHODS:The serum levels of TG, TC, LDL-C,HDL-C, apo AI, apo B and Lp(a) were measured and the polymorphism of the apo(a) PNR was studied by using PCR-SSCP in 153 random normal individuals in Hubei Han population respectively. RESULTS:The relative frequencies of apo(a) PNR allele were significantly different from western population. The apo(a) gene which copy number of PNR is 5 was associated with high Lp(a) levels. No marked differences in the levels of TG, TC,LDL-C, HDL-C, apo AI and apo B were found among the various genotype groups of apo(a) PNR in Hubei Han. CONCLUSION:The data of lipids and PNR polymorphism of the apo(a) gene from healthy Hubei Han were obtained. The apo(a) allele with 5(TTTTA)-repeats may be related to high serum Lp(a) levels in the Hubei Han population.     

Relationship between HDL subclass distribution and plasma glucose and lipid levels in patients with metabolic syndrome

AIM: To investigate high-density lipoprotein(HDL) subclass distribution and to analyze the relationship between HDL subclasses with plasma glucose and lipids in metabolic syndrome(MS). METHODS: Apolipoprotein A-I(apoA-I) contents of plasma HDL subclasses were determined by two-dimensional gel electrophoresis associated with immunodetection. The concentrations of lipids and apolipoproteins in the plasma were measured by an automated biochemical analyzer. RESULTS: Compared with the controls, the levels of fasting plasma glucose(FPG), total cholesterol(TC), triglyceride(TG), low-density lipoprotein cholesterol(LDL-C), LDL-C/high-density lipoprotein cholesterol(HDL-C), apolipoprotein B₁₀₀(apoB₁₀₀), apoB₁₀₀/apoA-I, systolic blood pressure(SBP), body mass index(BMI) and HDL_3b were increased in the MS patients(P<0.05). Meanwhile, HDL-C, apoA-I and preβ₂-HDL, HDL_2a and HDL_2b were decreased in the MS patients(P<0.01). With the increase in the plasma glucose level, the contents of HDL_2a and HDL_2b were decreased in the MS patients(P<0.05), while preβ₁-HDL was increased(P<0.05). With the decrease in the HDL-C level, the content of HDL_2b was decreased in the MS patients(P<0.01), while preβ₁-HDL was increased(P<0.01). With the increase in the TG level and the decrease in the HDL-C level, the content of HDL_2b had a decreasing trend and the content of small-particle preβ₁-HDL had an increasing trend, indicating that HDL maturation metabolism was disrupted. The correlation analysis showed that FPG was negatively correlated with the levels of HDL_2a and HDL_2b, HDL-C was negatively correlated with the level of preβ₁-HDL and positively correlated with the level of HDL_2b, and TG was positively correlated with the levels of preβ₁-HDL and HDL_3b. CONCLUSION: With the increases in the plasma glucose and TG, and the decrease in HDL-C in the MS patients, HDL particles have minifying tendency, and the maturation metabolism of HDL particles is disrupted.     

Relationship between 936C/T polymorphism in 3’-untranslated region of vascular endothelial growth factor gene and diabetic retinopathy in China

AIM: To study the distribution of 936C/T polymorphism in 3’- untranslated region of vascular endothelial growth factor (VEGF) gene in Chinese Han population and to analyze the relationship between the polymorphism and diabetic retinopathy (DR). METHODS: Two hundred and fifty-four patients with type 2 diabetes mellitus recruited in this study were divided into NPDR group (non-proliferative diabetic retinopathy), PDR group (proliferative diabetic retinopathy) and DM (diabetes without retinopathy) group. The normal control group consisted of 120 subjects. Genotypes were identified by PCR-RFLP among all the subjects, while other clinical parameters were measured. Serum levels of VEGF were tested by the method of ELISA. RESULTS: The frequencies of both genotype CC and allele C were significantly higher in NPDR group and PDR group than those in NC group (²=9.934, ²=4.899, P<0.05 and ²=10.895, ²=5.714, P<0.05) and DM group (²=7.490, ²=4.448, P<0.05 and ²=8.333, ²=5.227, P<0.05). However, the frequencies of genotype (TT+CT) and allele T were significantly lower in NPDR group and PDR group than those in NC group (²=9.934, ²=10.895, P<0.01 and ²=4.899, ²=5.714, P<0.05) and DM group (²=7.490, ²=8.333, P<0.01 and ²=4.448, ²=5.227, P<0.05). Multivariate logistic regression analysis showed that the levels of glycohemoglobin(HbA1c), total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C) and plasma VEGF presented a positive correlation with DR, respectively, and the 936C/T mutation of VEGF exhibited a negative correlation with DR (β=-1.027, OR=0.343, P<0.01, CI: 0.157-0.723). CONCLUSION: Allele 936C of VEGF may be a genetic marker susceptible to DR, while allele 936T may be a protective genetic marker of DR. The 936C/T mutation of VEGF may be a protective factor against DR.     

Effects of intranasal Escherichia coli on glucolipid metabolism in high-fat diet-induced obese mice

AIM: To study whether the pulmonary infection of Escherichia coli (E. coli) interferes the glucolipid metabolism in high-fat diet-induced obese mice. METHODS: High-fat diet-induced obese mice (n=48) and normal chow-fed control mice (n=48) were intranasally infused with 40 μL fluid containing 4×10⁹ CFUs E. coli. The serum, periepididymal adipose tissue and liver were obtained at 0 d, 1 d, 2 d, 3 d and 4 d after infection. The body mass, periepididymal adipose tissue and liver were weighed, and the levels of fasting blood glucose (FBG), fasting blood insulin (FINS), free fatty acid (FFA) and very-low-density lipoprotein (VLDL) were measured by ELISA. The serum total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C), and hepatic TG contents were detected, and the hepatic steatosis was observed under microscope with oil red O staining. RESULTS: Compared with day 0, the body mass, fat mass and fat index were decreased significantly from day 1 to day 4 after infection (P<0.05). The levels of FBG, FINS and HOMA-IR were apparently raised from day 2 to day 4 after infection (P<0.05). The contents of serum FFA, TG and VLDL were increased markedly from day 1 to day 4 after infection (P<0.05). However, the concentrations of serum TC, LDL-C and HDL-C were decreased obviously from day 1 to day 3 (P<0.05). The liver mass, liver index and TG content were significantly increased from day 1 to day 4 (P<0.05). Consistently, the lipid droplet accumulation in the liver cells was increased obviously at day 2 and day 4 after infection. Compared with control group, except the levels of serum TC, LDL-C and HDL-C in obese group substantially decreased, the other indexes were increased by different degrees during the whole experiment period (P<0.05). CONCLUSION: Pulmonary infection of Escherichia coli exacerbates the disorder of glucose and lipid metabolism in high-fat diet-induced obese mice, which contributes the development of insulin resistance and hepatic steatosis.     

Relationship between subclasses of serum HDL and LPL gene HindⅢ polymorphism in hyperlipidemia

AIM: To investigate lipoprotein lipase gene HindⅢ polymorphism and its relationship with serum lipids and apolipoprotein, serum HDL subclasses in patients with hyperlipoidemia. METHODS: Lipoprotein lipase gene HindⅢ polymorphism was assayed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The subclasses of serum HDL in 152 hyperlipoidemia patients and 128 healthy subjects were determined by two-dimensional gel electrophoresis conjunction with immunodetection method. RESULTS: H+H+ genotype and allele H+ in hyperlipoidemia and control groups were both the highest. In hyperlipidemia group, H+H+ genotypes tended to be higher than that in control group, while H+H- and H-H- genotypes were significantly lower (P<0.05). In hyperlipidemia group allele H+ carriers' frequency tended to be higher than that in control group (P<0.05). In hyperlipoidemia group, the genotype of H+H+ showed higher serum TG, apoB100 levels, TG/HDL-C ratio, preβ1-HDL, HDL3b and lower HDL2a, HDL2b compared with H-H- (P<0.05). In control group, the genotype of H+H+ had higher serum TG，HDL3c and lower HDL2a compared with H-H- (P<0.05). CONCLUSION: The HindⅢ polymorphism at intron 8 of LPL gene is associated with the general shift toward smaller size of HDL particle size in hyperlipoidemia, and the change of HDL subclasses distribution profile may be closely related to the pathogenesis of atherosclerosis in Chinese patients with hyperlipoidemia.     

Effect of ginsenoside Rb1 on liver cell pyroptosis in hyperlipidemia rats and its possible mechanism

AIM:To discuss the mechanism of ginsenoside Rb1 against liver lipid deposition by observing the effect of ginsenoside Rb1 on liver cell pyroptosis in hyperlipidemia rats. METHODS:Totally 32 healthy SPF rats were randomly divided into control group, model group, ginsenoside Rb1 group and simvastatin group. The rats in control group was given the basic feed, while the others were given high-fat diet. The rats in ginsenoside Rb1 group and simvastatin group were given corresponding drugs. The rats in control group and model group were intraperitoneal injected with equal volume of saline. Eight weeks later, the serum levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) were tested by the automatic biochemistry analyzer. The pathological changes of the liver tissues were observed with HE staining. The protein and mRNA expression levels of pyroptosis-related factors NLRP3, caspase-1, IL-1β, IL-18 and GSDMD were detected by Western blot and RT-qPCR. RESULTS:Compared with control group, the serum levels of TC, TG and LDL-C in model group were increased significantly (P<0.01), and the HDL-C content was decreased significantly (P<0.05). The steatotic liver cells covered the visual field. The mRNA and protein expression levels of NLRP3, caspase-1, IL-1β, IL-18 and GSDMD were increased significantly (P<0.01). Ginsenoside Rb1 significantly decreased the serum levels of TC, TG and LDL-C (P<0.05), and significantly increased the content of HDL-C (P<0.01). Ginsenoside Rb1 also significantly decreased the degree of steatosis, and the number and size of lipid droplets. The mRNA and protein expression levels of NLRP3, caspase-1, IL-1β, IL-18 and GSDMD were decreased significantly (P<0.05 or P<0.01). CONCLUSION:Ginsenoside Rb1 atte-nuates liver injury and inhibits liver lipid deposition in hyperlipidemia rats by reducing the expression of hepatic pyroptosis-related factors.     

Serum apelin and chemerin levels in female obese children and their correlation with insulin resistance

AIM: To investigate the serum levels of apelin and chemerin in female obese children and their correlation with insulin resistance (IR).METHODS: Thirty-five female children participated in the study, 20 of which were obese and 15 were non-obese controls ,without statistical difference in age between the 2 groups. Serum levels of apelin and chemerin were measured by ELISA method. The concentrations of triglyceride (TG), cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), fasting glucose and fasting insulin (FINS) were measured. Body mass index standard deviation score (BMI-SDS) and homeostasis model assessment of insulin resistance (HOMA-IR) were calculated for all participants. RESULTS: A significant difference of BMI between obese group and control group (24.02±3.90 vs 16.46±1.93, P<0.01) was observed. Serum levels of TG, LDL-C, FINS, and HOMA-IR were significantly higher in obese group than those in control group (allP<0.05). Serum levels of apelin and chemerin were also significantly higher in obese children than those in the controls . Serum level of apelin was positively correlated with BMI-SDS (r=0.356, P<0.05), TG (r=0.548, P<0.01), FINS (r=0.541, P<0.01) and HOMA-IR (r=0.551, P<0.01) in all individuals. The negative correlation between serum chemerin level and age (r=-0.362, P< 0.05), and positive correlations between serum chemerin level and BMI-SDS (r= 0.315, P<0.01), TG (r= 0.28, P<0.05), FINS (r= 0.38, P<0.01) and HOMA-IR (r= 0.41, P< 0.01) were detected.CONCLUSION: Increased serum apelin and chemerin levels are correlated with insulin resistance, indicating their roles in the pathogenesis of children obese.     

RXRα agonist bexarotene inhibits atherosclerosis in ApoE-/- mice by regulating TGF-β1/Smad2 signaling pathway

AIM To observe the effect of retinoid X receptor α （RXRα） agonist bexarotene （Bex） on the proliferation of transforming growth factor β1 （TGF-β1）-induced vascular smooth muscle cells （VSMCs） and atherosclerosis in apolipoprotein E knockout （ApoE^-/-） mice, and to explore the underlying mechanism. METHODS Ten C57BL/6 mice were selected as normal control group, and 30 ApoE^-/- mice were randomly divided into 3 groups： ApoE^-/- group, ApoE^-/-+Bex5 （5 mg·kg^-1·d^-1 Bex） group and ApoE^-/-+Bex10 （10 mg·kg^-1·d^-1 Bex） group. Bex was intragastrically given once a day for 8 weeks. The levels of triglyceride （TG） and total cholesterol （TC） were determined by oxidase method, and select masking method was used to determine serum levels of low-density lipoprotein cholesterol （LDL-C） and high-density lipoprotein cholesterol （HDL-C）. The protein levels of TGF-β1, p-Smad2 and Smad2 were determined by Western blot. HE staining was used to observe the intima of the thoracic aorta. The VSMCs were cultured with tissue patch method, and the proliferation of VSMCs was measured by BrdU incorporation method. RESULTS The serum levels of TG, TC and LDL-C, and the expression of TGF-β1 and p-Smad2 in thoracic aorta in ApoE^-/- group were significantly higher than those in C57BL/6 group （P<0.01）. Bex increased p-Smad2 protein level in thoracic aorta in a dose-dependent manner, inhibited the intimal plaque formation and vascular medial proliferation, and decreased the plaque area in ApoE^-/- mice （P<0.01）. No significant difference in serum levels of TG, TC, HDL-C and LDL-C, and TGF-β1 and Smad2 expression in thoracic aorta among ApoE^-/- group, ApoE^-/-+Bex5 group and ApoE^-/-+Bex10 group was observed. TGF-β1 （0.1~10 μg/L） promoted the proliferation of VSMCs, while Bex （10^-9~10^-7 mol/L） inhibited TGF-β1 （5 μg/L）-induced proliferation of VSMCs in a concentration-dependent manner. Bex （10^-7 mol/L） synergistically promoted the protein level of p-Smad2 in VSMCs induced by TGF-β1 （P<0.01）, but inhibited TGF-β1-induced nuclear translocation of p-Smad2. CONCLUSION RXRα agonist Bex inhibits the formation of atherosclerosis in ApoE^-/- mice, and its mechanism may be related to the regulation of TGF-β1/Smad2 pathway.     

Effects of Panax notoginseng saponins on PCSK9-LDLR expression and blood lipid metabolism in golden hamsters

AIM: To explore the regulatory effect of Panax notoginseng saponins (PNS) on hyperlipidemia in golden hamsters and its mechanism. METHODS: The hamsters (n=30) were randomly divided into 3 groups:normal group, model group, and PNS group. The animals in normal group was given common feed. The animals in other groups were given high-fat diet to construct a hyperlipidemia model. After induction for 4 weeks, the drugs were given by intraperitoneal injection for another 12 weeks. After the last drug given, the serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) were measured by biochemical tests. The distribution and expression of proprotein convertase subtilisin/kexin type 9 (PCSK9) and low-density lipoprotein receptor (LDLR) in liver were detected by real-time PCR, immunohistochemistry and Western blot. RESULTS: Compared with normal group, the serum levels of TC, TG, LDL-C and ALT in model group were increased significantly (P < 0.05). The expression of PCSK9 was increased, while the protein level of LDLR was decreased (P < 0.05). Compared with model group, the levels of TC, TG, LDL-C and ALT in PNS group were decreased significantly (P < 0.05), PCSK9 was mainly distributed in cytomembrane with decreased expression, and LDLR was mainly distributed in the cell membrane and plasma with increased expression. HDL-C and AST had no significant change during this time. CONCLUSION: Panax notoginseng saponins reduces the blood lipid levels in golden hamsters, which may be related to the regulation of PCSK9-LDLR signaling pathway.     

Resveratrol regulates iNOS to inhibit atherosclerosis in C57BL/6J mice

AIM: To investigate the effect of resveratrol on the lipids(CHOL, TG, LDL-C and HDL-C), nitric oxide(NO), peroxynitrite anion(ONOO^-) and the expression of inducible nitric oxide synthase(iNOS) in the artery of the mice with ovariotomy(OVX).METHODS: The lipid levels and NO level in the serum were measured. The changes of atherosclerosis were evaluated with Oil Red O staining. The expression of iNOS was measured by DAB staining and Western blot. The ONOO^- production was measured by DAB staining.RESULTS: Compared with sham group, the levels of the lipids and NO production in OVX+ high fat(HF) group were increased(P<0.05). Compared with OVX+HF group, the levels of the lipids and NO production in resveratrol group were decreased(P<0.05). Fourteen weeks later, the atherosclerosis model was successfully established. Compared with OVX+HF group, the iNOS expression and the ONOO^- production in resveratrol group were decreased(P<0.05), while those in sham group were increased(P<0.05).CONCLUSION: Resveratrol prevents and treats atherosclerosis by inhibiting the iNOS expression in C57BL/6J mice.     

Changes of liver lipid metabolism-related PPAR-γ/LXR-α/ABCG1 pathways and inflammatory factors in atherosclerotic mice and role of Huayu-Qutan recipe

AIM To observe the changes of liver lipid metabolism-related peroxisome proliferator-activated receptor-γ (PPAR-γ)/liver X receptor-α (LXR-α)/ATP-binding cassette transporter G1 (ABCG1) signaling pathways and inflammatory factors in mice with atherosclerosis (AS), and to investigate the effects of Huayu-Qutan recipe (HYQT) on hepatic lipid metabolism and inflammatory response and the mechanisms. METHODS ApoE^-/- mice (n=24) were randomly divided into model group, HYQT group and simvastatin group, and C57BL/6J mice (n=8) were used as control group. Except for the control group, the mice in other groups were given high-fat diet. After 12 weeks of modeling, the mice in HYQT and simvastatin groups were intragastrically given the corresponding drugs, and the mice in control and model groups were given the same volume of normal saline. After 8 weeks, the serum levels of triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) were measured by an automatic biochemical analyzer. HE and oil red O staining was used to observe liver histopathological and lipid changes. The hepatic levels of free fatty acid (FFA), TG, tumor necrosis factor-α (TNF-α), Toll-like receptor 4 (TLR4) and interleukin-1β (IL-1β) were detected by ELISA. The protein expression of PPAR-γ, LXR-α and ABCG1 was determined by Western blot. RESULTS Compared with control group, the serum levels of TC, TG and LDL-C in model group were significantly increased (P<0.01), and the HDL-C content was significantly decreased (P<0.01). Liver steatosis, cell size augmentation and lipid deposition were obvious, and liver FFA and TG levels were significantly increased (P<0.01). The liver levels of TLR4, TNF-α and IL-1β were significantly increased (P<0.01), while the protein expression of PPAR-γ, LXR-α and ABCG1 was significantly decreased (P<0.05 or P<0.01). Compared with model group, the serum levels of TC, TG and LDL-C in simvastatin group and HYQT group were significantly decreased (P<0.05 or P<0.01), and the HDL-C content was significantly increased (P<0.01). Liver steatosis was weakened, and liver lipid deposition and FFA and TG levels were significantly decreased (P<0.05 or P<0.01). The liver levels of TLR4, TNF-α and IL-1β were significantly decreased (P<0.01), while the protein expression of PPAR-γ, LXR-α and ABCG1 was significantly increased (P<0.05 or P<0.01). CONCLUSION Huayu-Qutan recipe may exert anti-AS effect by regulating liver PPAR-γ/LXR-α/ABCG1 pathways and attenuating liver TRL4-mediated inflammatory responses.     

Qishen-Yiqi dripping pills attenuate atherosclerosis by regulating Ca2+ homeostasis via TRPC1/STIM1 pathway

AIM:To explore the therapeutic effect of Qishen-Yiqi dripping pills (QS) on atherosclerosis (AS) and the mechanism. METHODS:AS rat model was established by high-fat diet, and SD rats were randomly divided into normal control group, AS model group, low-dose, middle-dose and high-dose QS groups, and positive group (n=6 each). After administration for 12 weeks, serum samples were collected to detect the serum lipid and Ca²⁺ levels. HE staining was used evaluated the histopathological changes of arterial tissue. The serum levels of interleukin-1β (IL-1β), IL-6 and tumor necrosis factor-α (TNF-α) were measured by ELISA. The nitric oxide (NO) level was detected by nitrate reductase method. The protein levels of transient receptor potential channel protein 1 (TRPC1), stromal interaction molecule 1 (STIM1) and endothelial NO synthase (eNOS) were determined by Western blot. RESULTS:QS significantly reduced the arterial damage via inhibiting the formation of atherosclerotic plaque and attenuated intimal thickening and vascular stenosis. Compared with AS group, the serum levels of total cholesterol (TC), triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C) were decreased significantly and the levels of high-density lipoprotein cholesterol (HDL-C) were increased significantly in high-dose QS group (P<0.05). The serum levels of IL-1β, IL-6 and TNF-α in high-dose QS group were lower than those in AS group (P<0.05). Compared with AS group, the serum Ca²⁺ level was lowered and the arterial tissue NO level was elevated in QS groups (P<0.05). Compared with AS rats, the protein levels of TRPC1 and STIM1 were decreased significantly and the protein level of eNOS was increased significantly in the rats treated with QS (P<0.05). CONCLUSION:QS regulate calcium homeostasis via TRPC1/STIM1 pathway, increase the production of NO and inhibit the inflammatory responses, thus exerting anti-AS effect.     

Association of rs2162459 polymorphism in CYP7A1 gene with effectiveness of atorvastatin in northern Chinese Han population

AIM：To investigate the association of the polymorphism of rs2162459 locus in cytochrome P-450, family 7, subfamily A, polypeptide 1 (CYP7A1) gene, encoding cholesterol 7α-hydroxylase, with the effectiveness of atorvastatin in northern Chinese Han population. METHODS：Clinical data and blood samples of 200 cases of hyperlipidemia patients were collected. The variants of rs2162459 in CYP7A1 gene were detected by multiplex SNaPshot technology. Several genetic models were constructed to analyse the association of gene polymorphism with the effectiveness of atorvastatin by logistic regression method. RESULTS：The polymorphism of rs2162459 was consistent with Hardy-Weinberg equilibrium. The baseline high-density lipoprotein cholesterol (HDL-C) concentrations in three genotypes of AA, GA and GG were (136±0.94) mmol/L, (1.16±0.38) mmol/L and (1.07±0.28) mmol/L, respectively (P<0.05). There were differences in the genotypic frequency and allelic frequency between atorvastatin effective and ineffective groups (both P<005). Significant differences in regulating HDL-C level among the three genotypes of rs2162459 were found after logistic regression. The results of additive model, generalized model and dominant model, presented as OR (95% CI), were 1.74 (1.09~2.77), 2.86 (1.13~7.25) and 2.21 (1.12~4.33), respectively. CONCLUSION: The baseline HDL-C level in the carriers of GG genotype is lower than that in the carriers of the other two genotypes. The HDL-C-elevating effect of atorvastatin on GG genotype carriers is more significant than that on AA genotype carriers.     

Changes of heart function and serum cardiac troponin I in early type 2 diabetic rats

AIM: To observe the changes of heart function and the expression of serum cardiac troponin I(cTnI) in early type 2 diabetic rats, and to explore the role of cTnI in the development of type 2 diabetes and early diabetic cardiomyopathy.METHODS: The type 2 diabetes rat model was established by an injection of streptozotocin after high fat diet(5 weeks). The rats were randomly divided into control group, model group of 2 weeks, and model group of 4 weeks. M-mode echocardiography was performed for echocardiographic measurements. Fasting blood glucose(FBG), total cholesterol(TC), triglyceride(TG), high density lipoprotein-cholesterol(HDL-C), low density lipoprotein- cholesterol(LDL-C), fasting insulin(FINS) and cTnI levels were tested. HE staining was used to observe the pathological changes of myocardial structures. The alteration of cTnI in myocardium was determined by Western blot.RESULTS: Compared with normal group, the levels of TC, TG and LDL-C in type 2 diabetic rats were significantly increased, HDL-C levels were significantly reduced. Cardiac histological analysis revealed that type 2 diabetes induced cardiomyocytes degeneration and necrosis. The expression of cTnI increased significantly in diabetic groups compared to control group, and that in model group of 4 weeks increased far more than that in model group of 2 weeks(P<0.05).CONCLUSION: The increased level of cTnI and the change of the heart function may be associated with the development diabetic cardiomyopathy. These changes are valuable for the early clinical diagnosis of myocardial injury in diabetic cardiomyopathy.     

Effects of gypenosides on PCSK9 gene expression and blood lipids lowered by simvastatin

AIM: To explore the effect of gypenosides (GPs) on PCSK9 gene expression in hyperlipidemic rat liver and the blood lipids lowered by simvastatin.METHODS: Healthy male SD rats (n=60) were randomized into 5 groups:normal control group, hyperlipidemic model group, simvastatin group, GPs group and GPs combined with simvastatin group (combined group). The rats in all groups were fed high-fat diet except normal control group which were fed with ordinary diet. The rats in control group and hyperlipidemic model group were gavaged with 0.3% CMC-Na every day. The rats in GPs group were gavaged with GPs at 160 mg·kg^-1·d^-1. The rats in simvastatin group were gavaged with simvastatin at 5 mg·kg^-1·d^-1. The rats in combined group were gavaged with GPs and simvastatin. The experiment lasted for 8 weeks. The rats were anesthetized with chloral hydrate, and abdominal arterial blood samples were collected to detect the total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C). The body weight and the wet weight of the livers were measured, and the liver index was calculated. The pathological changes of the livers were observed under microscope with HE staining. The expression of PCSK9 and low-density lipoprotein receptor (LDLR) at mRNA and protein levels was determined by real-time PCR and Western blot. RESULTS: The model of hyperlipidemia rats was established successfully. Compared with model group, the levels of TC, TG and LDL-C in simvastatin group, GPs group and combined group were obviously decreased (P<0.05), and the HDL-C levels were obviously upregulated (P<0.05). Compared with model group, the liver indexes in simvastatin group, GPs group and combined group were obviously decreased (P<0.05). The pathological changes of the liver tissues showed that hepatic adipose appeared in model group, and that in simvastatin group and GPs group had different degrees of relief, especially in combined group. Compared with model group, the mRNA expression levels of PCSK9 and LDLR in simvastatin group were obviously increased, while the mRNA expression levels of PCSK9 in GPs group and combined group were obviously decreased (P<0.05), and the mRNA expression of LDLR in combined group was obviously increased (P<0.05). Compared with model group, the protein expression of PCSK9 and LDLR in simvastatin group was obviously increased, while the protein expression levels of PCSK9 in GPs group and combined group were obviously reduced, and the LDLR protein levels were obviously increased (P<0.05). CONCLUSION: Gypenosides inhibit the expression of PCSK9 and increase the expression of LDLR in the liver. The combination of gypenosides and simvastatin promotes the lipid-lowering effect of simvastatin and attenuates hepatic steatosis, which may be related to inhibiting the expression of PCSK9 in the liver.     

Effect of tanshinone ⅡA on liver lipid deposition and ferroptosis-related protein expression in ApoE-/- mice

AIM To observe the effect of tanshinone ⅡA on liver lipid deposition and ferroptosis-related protein expression in ApoE^-/- mice. METHODS Thirty-two ApoE^-/- mice were randomly divided into model group, high-dose (60 mg/kg) tanshinone ⅡA group, low-dose (30 mg/kg) tanshinone ⅡA group and simvastatin group, and C57BL/6J mice (n=8) were used as normal control group. The mice in normal control group were given the basic feeding, while the others were given high-fat diet. The mice in tanshinone ⅡA groups and simvastatin group were given corresponding drugs. The mice in normal control group and model group were intraperitoneally injected with equal volume of saline. Eight weeks later, the serum levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) were tested by automatic biochemistry analyzer. The liver tissues were stained with HE and oil red O. The contents of reactive oxygen species (ROS) and glutathione (GSH) in liver tissues of the mice were measured by commercially available kits. The liver glutathione peroxidase 4 (GPX4) and p53 were detected by immunohistochemical method. The protein and mRNA expression levels of ferroptosis-related factors GPX4, xCT/SLC7A11, p53 and ferritin heavy chain 1 (FTH1) were determined by Wes automatic Western blot quantitative analysis system and RT-qPCR. RESULTS Compared with normal control group, the serum levels of TC, TG and LDL-C in model group were increased significantly (P<0.05 or P<0.01), and HDL-C did not change significantly. The fat vacuoles were clearly visible in liver tissue. The content of ROS in liver tissue was increased significantly,and GSH was decreased significantly (P<0.01). The mRNA and protein expression levels of p53 were increased significantly, and GPX4, xCT/SLC7A11 and FTH1 were decreased significantly (P<0.05 or P<0.01). Compared with model group, tanshinone ⅡA significantly decreased the serum levels of TC, TG and LDL-C (P<0.05 or P<0.01), and HDL-C did not change significantly. High-dose and low-dose tanshinoneⅡA also significantly decreased the degree of steatosis, and the size of lipid droplets. The content of ROS in liver tissues was decreased significantly, and GSH was increased significantly (P<0.01). The mRNA and protein expression levels of GPX4, xCT/SLC7A11 and FTH1 were increased significantly, and p53 were decreased significantly (P<0.05 or P<0.01). CONCLUSION Tanshinone ⅡA reduces liver lipid deposition and lipid peroxidation damage in ApoE^-/- mice, which may be related to the intervention of ferroptosis-related proteins in the liver cells.     

Serum levels of visfatin and tumor necrosis factor-alpha in patients with pre-eclampsia and their relationship with insulin resistance

AIM: To explore the serum levels of visfatin (VF) and tumor necrosis factor-alpha (TNF-α) in the patients with pre-eclampsia (PE) and their correlation with insulin resistance (IR). METHODS: The severe PE patients (n=30), mild PE patients (n=30) and normal pregnant women (n=40) were selected according to the classification standard of PE. The serum levels of VF and TNF-α were measured by ELISA. Fasting plasma glucose (FPG) and fasting insulin (FIns) were detected by glucose oxidase method and radioimmunoassay, respectively. Triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) were measured by an automatic biochemical analyzer. According to calculating the mean arterial pressure (MAP), body mass index (BMI) and homeostatic model assessment for insulin resistance index (HOMA-IR), the correlation between IR and the levels of serum VF as well as TNF-α were analyzed.RESULTS: The levels of VF and TNF-α in severe PE group and mild PE group were significantly lower than those in normal pregnancy group (P<0.05). In addition, the levels of VF and TNF-α in severe PE group were lower than those in mild PE group (P<0.05). Linear correlation analysis showed that serum VF was positively correlated with TNF-α and HDL-C (P<0.05), and negatively with MAP and FIns (P<0.05). The serum TNF-α was positively correlated with HDL-C (P<0.05), and negatively with BMI, TG, MAP and FIns (P<0.05). Multiple stepwise regression analysis showed that FBG, FIns and HOMA-IR were relative independent factors of serum VF and TNF-α (P<0.05).CONCLUSION: Serum levels of VF and TNF-α are closely related to IR.     

Atorvastatin inhibits atherogenesis by RXRα-mediated depressing oxidative stress in STZ-induced diabetic ApoE-/- mice with fat-rich diet

AIM: To explore the effects of atorvastatin (Atorv) on atherosclerosis in streptozotocin (STZ)-induced diabetic apolipoprotein E knockout (ApoE-/-) mice with fat-rich diet and the possible mechanism. METHODS：C57 mice served as control. ApoE-/- mice (n=34) fed with high-fat diet were randomly divided into ApoE-/- group, STZ-ApoE-/- group and STZ-ApoE-/-+Atorv group. Intraperitoneal injection of streptozotocin was performed to create diabetic animal model. Blood glucose was determined by glucose oxidase method. Blood lipid levels were detected by enzymic method or selective homogeneous method. The plaque area in the thoracic aorta was measured by HE staining. The protein level of nicotinamide-adenine dinucleotide phosphate (NADPH) oxidase subunit gp91phox in the thoracic aorta was determined by Western blotting. The levels of reactive oxygen species (ROS) in blood and thoracic aorta homogenates were detected by Fenton reaction and Griess reagent. Human umbilical vein endothelial cells (HUVECs) were isolated from healthy umbilical cords by collagenase I and cultured. ROS production was detected by flow cytometry. NADPH oxidase activity was measured using lucigenin assay.Effects of retinoid X receptor α (RXRα) on inhibition of oxidative stress by atorvastatin were evaluated by RNA interference and plasmid transfection. RESULTS：(1) Compared with C57 group, the plaque areas of the thoracic aorta in ApoE-/- group were increased. No difference of the fasting glucose between the 2 groups was observed. The levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), thoracic aorta gp91phox protein and ROS in blood and thoracic aorta homogenates were higher in ApoE-/- group than those in C57 group. (2) Compared with ApoE-/- group, the plaque areas of the thoracic aorta in STZ-ApoE-/- group were further enlarged ［(314.13±35.72) μm2 vs (215.88±34.19) μm2, P<0.05］. The levels of blood glucose, TG, TC and LDL-C, thoracic aorta gp91phox protein and ROS in blood and thoracic aorta homogenates were higher in STZ-ApoE-/- group than those in ApoE-/- group (P<0.05). (3) Compared with STZ-ApoE-/- group, the plaque areas of the thoracic aorta in STZ-ApoE-/- +Atorv group were reduced ［(217.47±24.56） μm2 vs （314.13±35.72） μm2, P<0.05］. The levels of blood glucose, LDL-C, TC, HDL-C and TG showed no significant difference between the 2 groups. Thoracic aorta gp91phox protein level and ROS production in blood and thoracic aorta homogenates were lower in STZ-ApoE-/- +Atorv group than those in STZ-ApoE-/- group (P<0.05). (4) High glucose-induced increases in NADPH oxidase activity and gp91phox expression were significantly inhibited by atorvastatin (10-6 mol/L) in HUVECs. The inhibitory effects of atorvastatin on high glucose-induced ROS production and NADPH oxidase activation were largely impaired when the cells were transfected with RXRα siRNA. However, the effect of atorvastatin was significantly strengthened when RXRα was over-expressed in the HUVECs transfected with RXRα plasmid. CONCLUSION：Atorvastatin inhibits atherogenesis by depressing high glucose-induced oxidative stress in diabetic ApoE-/- mice with fat-rich diet. The anti-oxidative stress effect of atorvastatin is mediated by RXRα.