Serum levels of inflammatory factors and adiponectin in type 2 diabetic retinopathy patients

AIM: To investigate the serum levels of inflammatory factors and adiponectin in type 2 diabetic retinopathy (DR) patients.METHODS: One hundred and ten cases of type 2 diabetic patients were divided into 3 groups: no diabetic retinopathy group (DM, n=35), non-proliferative diabetic retinopathy group (NPDR, n=45), and proliferative diabetic retinopathy group (PDR, n=30). Other 40 normal persons served as controls (NC group). The physical examinated was performed for each patient. Serum levels of fasting plasma glucose (FPG), glycated hemoglobin A1c (HbA1c), 2 h postprandial plasma glucose (2hPG), fasting insulin (FINS), total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), adiponectin, intercellular adhesion molecule-1(ICAM-1), tumor necrosis factor-α (TNF-α) and high-sensitivity C-reactive protein (hs-CRP) were measured. Insulin resistance index (HOMA-IR) was also calculated.RESULTS: The systolic blood pressure, body-mass index, waist-hip ratio, serum levels of TG, LDL-C, FPG, 2hPG, HbA1c, ICAM-1, TNF-α, hs-CRP and HOMA-IR were significantly higher in DM group, NPDR group and PDR group than those in NC group (P<0.05). The systolic blood pressure, serum levels of ICAM-1, TNF-α, hs-CRP and HOMA-IR were higher in NPDR group and PDR group than those in DM group (P<0.05). The serum concentration of adiponectin was lower in DM group, NPDR group and PDR group than that in NC group (P<0.05), and that was also lower in NPDR group and PDR group than that in DM group (P<0.05). The negative correlations between adiponectin and ICAM-1 (r=-0.735,P<0.01), TNF-α (r=-0.781,P<0.01), hs-CRP (r=-0.768, P<0.01) or HOMA-IR (r=-0.752, P<0.01) were observed. The relationships between HOMA-IR and ICAM-1 (r=0.857,P<0.01), TNF-α (r=-0.906, P<0.01) or hs-CRP (r=-0.888,P<0.01) were positive.CONCLUSION: The results suggest that inflammatory refactors and adiponectin play important roles in the pathophysiology and progression of DR. The protective effects of adiponectin on DR may be related with its anti-inflammatory reactions to improve insulin resistant.     

GLP-1 down-regulates mRNA expression of SOCS-3 and SREBP-1c in rats with nonalcoholic fatty liver disease

AIM: To investigate the effects of glucagon-like peptide-1(GLP-1) on mRNA expression of SOCS-3 and SREBP-1c in the rats with nonalcoholic fatty liver disease. METHODS: Male SD rats were randomly divided into normal control(NC) group, high fat(HF) group and HF+liraglutide(Lira) group. The rats in HF group and HF+Lira group were given high-fat diet for 16 weeks. After 12 weeks of high-fat diet feeding in HF+Lira group, Lira(600 μg·kg^-1·d^-1) was intraperitoneally injected for 4 weeks. At the end of the 16th week, the rats were killed. The pathological changes of the liver were observed under optical microscope. The serum levels of alanine aminotransferase(ALT), aspartate aminotransferase(AST), triglyceride(TG) and total cholesterol(TC) were detected by automatic biochemical analyzer. TG contents of liver were measured by GPO-PAP method. The fasting insulin(FINS) was determined by ELISA, and insulin resistance index was assessed by homeostasis mode assessment(HOMA-IR). The mRNA expression of SOCS-3 and SREBP-1c in the liver tissues was detected by RT-qPCR. RESULTS: Compared with NC group, HOMA-IR, TG of liver, and the serum levels of ALT, AST, TG, TC and FINS in HF group were obviously increased(P<0.01). Compared with HF group, HOMA-IR, TG of liver, and the serum levels of ALT, AST, TG, TC and FINS in HF+Lira group were all obviously decreased(P<0.05 or P<0.01). The mRNA expression of SOCS-3 and SREBP-1c in HF group was significantly higher than that in NC group(P<0.01). The mRNA expression of SOCSV3 and SREBP-1c in HF+Lira group was significantly decreased as compared with HF group(P<0.05). CONCLUSION: Liraglutide may improve the IR and reduce TG of liver through decreasing the mRNA expression of SOCS-3 and SREBP-1c, so as to play a therapeutic role in nonalcoholic fatty liver disease.     

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.     

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.     

Effects of liraglutide on adiponectin and insulin resistance in rats with non-alcoholic fatty liver disease

AIM：To investigate the effects of glucagon-like peptide 1 analog, liraglutide, on adiponectin and insulin resistance in the rats with diet-induced non-alcoholic fatty liver disease (NAFLD). METHODS：Male rats were randomly divided into 3 groups:normal diet (ND) group (n=10), high-fat diet (HFD) group (n=10), and HFD with intraperitoneal injection of liraglutide group (n=10, first 12 weeks with HFD, later 4 weeks with liraglutide). All treatments continued for 16 weeks, and then the rats were killed ethically and the blood samples and liver tissues were collected. The levels of alanine aminotransferase (ALT), fasting blood glucose (FBG), total cholesterol (TC) and triglyceride (TG) were detected by a biochemical automatic analyzer. The levels of free fatty acids (FFAs), fasting insulin (FINS) and adiponectin were measured by RIA and ELISA. RESULTS：Compared with HFD group, the body weight, liver index, homeostasis assessment-insulin resistance (HOMA-IR), the serum levels of TG, TC, ALT and FBG, and the liver levels of TG, TC and FFAs in the rats in liraglutide group were apparently lower, the degree of hepatic steatosis and inflammatory activity significantly decreased (P<0.05), and the level of adiponectin in the serum and liver homogenate increased ob-viously (P<0.05). The level of adiponectin in the liver homogenate was negatively correlated with the levels of FFAs in the liver homogenate. CONCLUSION:Liraglutide is beneficial for NAFLD rats to improve insulin resistance and reduce hepatic steatosis by increasing the level of adiponectin in the serum and liver tissues.     

Vaspin and insulin signaling pathways in obstructive sleep apnea-hypopnea syndrome animal model

AIM: To establish a suitable chronic intermittent hypoxia (CIH) model in rats for modeling the obstructive sleep apnea-hypopnea syndrome (OSAHS) and further to explore the relationship between vaspin and insulin signaling pathways at mRNA and protein levels. METHODS: Healthy male Wistar rats were divided into control group and CIH group. The rats in control group were raised under physiological condition, and the rats in CIH group were kept in the plexiglass chamber between 9:00~17:00 and underwent intermittent hypoxic challenge 8 h/d for 8 weeks. The blood pressure of arteria caudilis in the rats was measured by tail cannulation before and after study. Fasting plasma glucose (FPG), total cholesterol (TC), triglycerides (TG), fasting insulin (FINS), vaspin and adiponectin levels were measured. The mRNA expression of vaspin was detected by real-time PCR. The protein levels of vaspin, Akt and p-Akt were determined by Western blotting.RESULTS: Before the experiment, the weight and blood pressure of the rats had no significant difference, while after the experiment the blood pressure in CIH group was obviously higher than that in control group. FPG, FINS, TG and TC in CIH group were also markedly higher than those in control group (P<0.05). Serum vaspin concentration was significantly correlated with FPG, FINS, HOMA-IR and TC. The expression of vaspin at mRNA and protein levels in the fat tissue of CIH group was evidently higher than that in control group. The protein levels of p-Akt decreased distinctly in CIH group. CONCLUSION: The levels of FINS, HOMA-IR, SaO2 and vaspin were markedly higher than those in control group, indicating that there was a close relationship between vaspin and insulin resistance in OSAHS.     

Hypolipemic and hypoglycemic effects of total triterpenoids from Psidium guajava leaves on type 2 diabetic rats

AIM: To investigate the effects of total triterpenoids from Psidium guajava leaves (TTPGL) on blood glucose and lipids in type 2 diabetic rats. METHODS: The diabetic rats were induced by intraperitoneal injection of streptozotocin at dose of 35 mg/kg and feeding with high-fat diet. The animals were divided into 5 groups: diabetic model control group (model), TTPGL treatment groups (with the doses of 60, 120 and 240 mg/kg, respectively) and rosiglitazone treatment group (3 mg/kg). Another 12 normal SD rats were used as the normal controls. The rats received daily treatment for 6 weeks, and then the levels of fasting blood glucose (FBG), fasting insulin (FINS), triglyceride (TG), total cholesterol (TCH), free fatty acid (FFA), glycosylated hemoglobin (GHb) and glycosylated serum proteins (GSP) were measured. The protein expression of peroxisome proliferator-activated receptor γ (PPARγ) in adipose tissues was detected by Western blotting. RESULTS: Compared with normal control group, the levels of FBG, GHb and blood lipids were increased in type 2 diabetic rats. The FINS, insulin sensitivity index, and the protein expression of PPARγ in adipose tissues were decreased. Compared with model group, the levels of FBG and GSP were decreased,and the FINS, insulin sensitivity index, and the protein expression of PPARγ in adipose tissues significantly increased in TTPGL treatment groups (with the doses of 120 and 240 mg/kg). The levels of serum TG,TCH and FFA were significantly lower in TTPGL treatment groups (P<0.01 or P<0.05) as compared with the model controls. CONCLUSION: TTPGL decreases the levels of blood glucose and lipids in diabetic rats. TTPGL also increases serum insulin level and improves insulin sensitivity. The action mechanism of TTPGL may be related to the increase in the protein expression of PPARγ.     

Influence of Tangshenfang on diabetic liver injury and the expression of SIRT1 and PGC-1α in the liver tissue

AIM: To observe the effect of Tangshenfang (TS) on the liver protection and the levels of silent information regulator 1 (SIRT1) and peroxisom proliferator-activated receptor γ coactivator-1α (PGC-1α) in the liver tissue. METHODS: The rat model of diabetes mellitus (DM) was established by intravenous injection of streptozotocin (STZ;30 mg/kg) after having the high fat/high glucose diets for 1 month. The diabetic rats were randomly divided into DM group, DM with high-dose TS (TS^Hi) group, medium-dose TS (TS^Med) group and low-dose TS (TS^Low)group. The normal rats were served as control group. There were 8 rats in each group. After treatment with TS for 12 weeks, the serum biochemical indices including fasting blood glucose (FBG), triglyceride (TG), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were tested. Fasting insulin (FINS) was also detected by radioimmunoassay, and homeostatic model assessment for insulin resistance (HOMA-IR) was calculated. The serum levels of tumor necrosis factor-α (TNF-α) and interleukin-1 (IL-1) were measured by ELISA. The activity of SOD and content of MDA in the liver tissues were measured by the methods of hydroxylamine and thiobarbituric acid. The liver pathological changes were observed under light microscope with HE and Masson staining. The protein expression of SIRT1and PGC-1α in the liver tissues was determined by Western blot. RESULTS: In DM group, serum FBG, TG, ALT, AST, FINS, HOMA-IR, TNF-α and IL-1 were obviously increased compared with the control group (P<0.01). The fatty changes, local necrosis, inflammation and fibrosis in the liver tissues were observed. The content of MDA in liver increased, while the activity of SOD decreased markedly. The protein expression of SIRT1 and PGC-1α was decreased (P<0.05). In TS treatment groups, all these changes in DM rats were markedly reversed by TS, and the protein expression of SIRT1 and PGC-1α in the liver tissues was markedly increased. CONCLUSION: TS may protect the rats from diabetic liver injury by increasing the expression of SIRT1 and PGC-1α, and thereby improving insulin resistance and oxidative stress.     

Hypothalamic expression of orexin A and lipid metabolism disorder in rats with alimentary obesity

AIM: To evaluate the effect of orexin A in rat hypothalamus on lipid metabolism disorder in rats with alimentary obesity induced by high-fat diet.METHODS: The rat model of alimentary obesity was induced by high-fat diet. The levels of insulin, triglyceride (TG) and total cholesterol (TC) in the serum were detected by luminescent immunoassay and enzymic method. The mRNA expression of orexin A in rat hypothalamus was determined by real-time PCR.RESULTS: There were statistically significant differences of weight, body fat content, and Lee's index between high-fat diet group and control group after 8-week feeding of high-fat diet. Compared to control animals, the levels of insulin, TG and TC in the rats with alimentary obesity significantly increased by 50％, 94％ and 43％, respectively (P<0.05). The expression of orexin A in rat hypothalamus significantly decreased by 57％, and had significant negative correlation with Lee's index, insulin, TG and TC. Their correlation coefficients were r=-0.798 (P<0.05), r=-0.868 (P<0.05), r=-0.981(P<0.05) and r=-0.815 (P<0.05), respectively. CONCLUSION: Alimentary obesity and lipid metabolism disorder induced by high-fat diet are correlated with down-regulation of orexin A expression in rat hypothalamus.     

Serum lipid levels and ApoB gene Xba I and 3'-VNTR polymorphisms in longevous elderly Han population in Zhejiang

AIM: To study the levels of serum lipids and the effects of apolipoprotein B (ApoB) gene Xba I-restriction fragment length polymorphism (RFLP) and 3'-variable-number tandem repeat (VNTR) polymorphism on serum lipid and apolipoprotein levels in longevous elderly Han population in Zhejiang. METHODS: Physical and laboratory examinations were performed on longevous elders, children of longevous elders, normal control people and children of non-longevous elders, and the Xba I-RFLP and 3'-VNTR polymorphism in ApoB gene were genotyped. RESULTS: The serum levels of triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and ApoB were negatively correlated with longevity (P<0.05), while the level of high-density lipoprotein cholesterol (HDL-C) was positively correlated with longevity (P<0.01). The levels of total cholesterol (TC), LDL-C and ApoB were higher and the HDL-C level was lower in the people with Xba I-RFLP X⁺X^- genotype than those in the people with Xba I-RFLP X^-X^- genotype (P<0.01). The levels of TC, TG, LDL-C and ApoB were higher and the HDL-C level was lower in the people with 3'-VNTR large allele than those in the people with 3'-VNTR minor allele (P<0.01). CONCLUSION: High TG, LDL-C and ApoB levels and low HDL-C level are not conductive to longevity. Serum lipid levels in the people with Xba I-RFLP X^-X^- genotype or 3'-VNTR minor allele are healthy and conductive to longevity.     

Correlation among plasma leptin,resistin an type 2 diabetes mellitus

AIM：To assess the association of resistin,obesity,serum lipid levels and insulin resistance with plasma leptin.METHODS：The concentrations of fasting serum glucose,insulin,lipid profiles,plasma resistin and leptin were assayed in 80 cases (including 37 controls with normal glucose tolerance and 43 patients with type 2 diabetes mellitus).RESULTS：Fasting plasma leptin level was positively correlated with sex,body mass index (BMI),waist circumference (WC),waist-hip ratio and fasting serum insulin (F Ins) （P<0.01）.Fasting plasma leptin level was negatively correlated with insulin sensitivity index (r=-0.373,P<0.01).There was no correlation between the concentrations of plasma leptin and FPG,TG,TC and resistin （P>0.05）.CONCLUSION：Fasting plasma leptin level is positively correlated with obesity and insulin resistance,not resistin.Leptin may play a role in the pathogenesis of type 2 diabetes mellitus.     

Relationship between the E-selectin and oxidative stress in patients with obese type 2 diabetes mellitus

AIM: To investigate the relationship between the levels of soluble E-selectin and oxidative stress in patients with obese type 2 diabetes mellitus.METHODS: The level of E-selectin, the contents of ox-LDL and malondialdehyde (MDA) and activities of superoxide dismutase (SOD) were measured in patients with obese and non-obese type 2 diabetes mellitus.RESULTS: The levels of E-selectin, ox-LDL and MDA were higher in patients with obese type 2 diabetes mellitus than those in control group (P<0.05), and the contents of HDL-C, HDL2-C and HDL3-C were significantly lower than those in control group (P<0.01).The activity of SOD in patients with obese type 2 diabetes mellitus was significantly lower than that in control group.The contents of E-selectin and MDA were more markedly elevated in patients with obese type 2 diabetes mellitus than those in patients with non-obese type 2 diabetes mellitus (P<0.01,P<0.05) and the activity of SOD was also significantly lower than that in patients with non-obese type 2 diabetes mellitus (P<0.01).There was significantly positive correlation between E-selectin and HbA1c, waist circumference, TC, ox-LDL, MDA (r=0.352, P<0.05;r=0.634, P<0.05;r=0.517, P<0.05;r=0.480, P<0.05;r=0.572, P<0.05), and negatively correlation between E-selectin and HDL3-C (r=-0.374, P<0.05).CONCLUSION: The plasma level of E-selectin is markedly elevated in patients with obese type 2 diabetes mellitus.E-selectin is possibly associated with oxidative stress.     

Influences of apolipoprotein CII concentrations on high-density lipoprotein subclass distribution

AIM: To investigate the influence of serum apolipoprotein (apo) CII concentrations on the distribution of serum high-density lipoprotein (HDL) subclasses. METHODS: Serum HDL subclasses in 247 subjects were determined by two dimensional gel electrophoresis-immunodetection. RESULTS: With the increase in serum apolipoprotein CII levels, age, BMI, the contents of TG, TC, apoB100, apoCII, apoCIII, apoE, preβ1-HDL, preβ2-HDL, HDL3b and HDL3a increased significantly, but the contents of HDL-C, HDL2a and HDL2b decreased remarkably. The contents of preβ1-HDL increased with the rise in apoCII and apoA I levels, whereas the content of HDL2b increased with the rise in serum apoA I level in the same apoC II group, but decreased with the increase in serum apolipoprotein CII level in the same apoA I group. With the increase in the ratio of apoCII/ apoCIII, the content of preβ1-HDL elevated, but the content of HDL2b decreased. The correlation analysis illustrated that the apoCII level was positively correlated with preβ1-HDL (r=0.186, P<0.01), but inversely correlated with HDL2b (r=-0.149, P<0.05). The apoA I level was positively associated with all HDL subclasses (r in the range of 0.349-0.587, P<0.01). In addition, the apoCIII level was positively correlated with preβ1-HDL (r=0.184, P<0.01) and preβ2-HDL (r=0.178, P<0.01), while the apoE level was positively correlated with HDL3a (r=0.040, P<0.05). The apoB100 level was inversely correlated with HDL2a (r=-0.102, P<0.05). CONCLUSION: The particles of HDL show a general shift towards smaller size with the increase in apoCII levels, indicating that the maturation of HDL is abnormal. Whereas the contents of apoA I level correct the effect of apoCII on the distribution profile of HDL subclasses. The ratio of apoCII/apoCIII might also been taken as one of the indexes reflecting the distribution profile of serum HDL subclasses.     

Protective effect of chrysin on mice with insulin resistance

AIM: To explore the effects of chrysin on insulin resistance (IRe) in a mouse model. METHODS: Male C57 mice were randomly divided into control group, IRe group, low-dose chrysin group (IRe+chrysin-low) and high-dose chrysin group (IRe+chrysin-high). After 24 weeks, the body weight, liver index and fat mass in all mice were detected. The blood glucose, insulin level and HOMA-IR were measured to determine the changes of the insulin resistance in the animals. The oxidative stress (SOD, GSH-Px and MDA) was also measured. The mRNA expression of insulin signaling pathway molecules (IR, IRS1, IRS2, Glut2 and Glut4) and inflammatory factors (TNF-α, IL-1β, IL-6 and NF-κB) was analyzed by real-time PCR. The protein levels of IRS1 and p65, and their phosphorylation were detected by Western blot. RESULTS: After 24-week intervention, the indicators in IRe group were higher than those in control group, including body fat deposition, serum glucose, serum insulin, HOMA-IR and liver oxidative stress (P<0.01), indicating that the model of insulin resistance was successfully established. Low dose and high dose of chrysin decreased the body weight, serum glucose, serum insulin and HOMA-IR in the IRe mice (P<0.05). The liver oxidative stress was also reduced in both groups (P<0.05). However, no statistical difference of the indexes between IRe+chrysin-low group and IRe+chrysin-high group was observed. Chrysin upregulated the mRNA expression of IR, IRS1, IRS2, Glut2 and Glut4 (P<0.05), and down-regulated the mRNA expression of various inflammatory factors. The inhibitory effect of chrysin on the mRNA expression of NF-κB was observed (P<0.05), especially in high dose group (P<0.05). It was confirmed that the effect of chrysin on liver IRe was related with the increase in the p-IRS1 levels and decrease in the p-p65 levels by Western blot. CONCLUSION: Chrysin inhibits obesity, hyperglycemia and hyperinsulinemia, and relieves insulin resistance and oxidative stress, which might be closely related to the regulation of insulin signaling pathway and the inhibition of inflammatory factor expression.     

Role of apelin in rat myocardial ischemic preconditioning and ischemia/ reperfusion injury

AIM: To study the alteration and role of apelin in myocardial ischemic preconditioning and ischemia/reperfusion injury of rats.METHODS: Forty-five SD rats were randomly divided into three groups: ischemia/reperfusion group (IR),ischemia pre-conditioning group (IP) and sham operation group.ECG was continuously used to evaluate the score of arrhythmias.The protein levels of apelin-36 in myocardium and plasma were detected by radioimmunoassay.The expression of apelin was observed by immunohistochemistry.RESULTS: (1) The scores of arrhythmias in IP group (2.1±0.5) was only 58.3% of IR group (3.6±0.8) ( P<0.01).(2) The apelin-36 protein level of plasma and myocardium in IR group were respectively lower by 36.1% and 45.6% than those in SH group (P<0.01),and those in IP group were lower by 23.8% and 24.7% than those in SH group (P<0.01),but higher than those in IR group (18.9% and 38.5%,respectively,P<0.05).(3) The staining absorbance of apelin in IR,SH and IP group was (7.87±2.41),(22.53±2.54) and (14.23±2.15),respectively.There were significant differences between IR and SH (P<0.01) and between IP group and SH group (P<0.05).(4) The scores of arrhythmias in IP and IR were negatively correlated with the protein level of apelin-36 in myocardium (r= 0.847,P <0.01).CONCLUSION: The down-regulation of apelin-36 in the plasma and myocardium of rats indicates that apelin has an important role in myocardial ischemic preconditioning and ischemia/reperfusion injury.     

Oxymatrine ameliorates high fat-induced insulin resistance in ApoE-/- mice

ATM: To investigate the effect of oxymatrine (OXY) on high fat-induced insulin resistance in mice, and to investigate the mechanism. METHODS: ApoE^-/-mice with high-fat diet for 16 weeks were divided into insulin resistance group, and OXY groups at concentrations of 25, 50 and 100 mg/kg. C57BL/6J mice served as normal control group. The mice in OXY groups were gavaged with OXY for 8 weeks. Glucose tolerance test in the mice was performed. Fasting blood glucose (FBG), total cholesterol (TC), triglyceride (TG), fatty acid (FFA) and fasting insulin (FINS) in the plasma were measured. The mRNA expression of insulin receptor (INSR), insulin receptor substrate-2 (IRS-2), glucose transporter 2 (GLUT2) in the liver tissues was examined by RT-qPCR. The protein levels of GLUT2, INSR, IRS-2, p-INSR, p-IRS-2, PI3K, p-PI3K, serine/threonine protein kinase (AKT) and p-AKT were examined by Western blot.RESULTS: OXY reduced the levels of FBG, TC, TG, FFA and FINS, and attenuated insulin resistance. Compared with insulin resistance group, the mRNA expression of INSR, IRS-2 and GLUT2 significantly increased in OXY groups (P<0.05). The protein levels of p-INSR/INSR, p-IRS-2/IRS-2, p-PI3K/PI3K, p-AKT/AKT and GLUT2 also increased in OXY groups (P<0.05).CONCLUSION: OXY ameliorates high fat-induced insulin resistance in mice via PI3K/AKT pathway.     

PPARs signaling pathway is involved in diabetic hepatopathy in mice

AIM: To investigate the role of peroxisome proliferator-activated receptors (PPARs)-inflammation signaling pathways in diabetic hepatopathy. METHODS: Diabetic mouse model was established by feeding the mice with a high-energy diet for 4 weeks combined with intraperitoneal injection of streptozotocin (STZ; 40 mg·kg^-1·d^-1 for 5 d). The hepatopathy model was confirmed by histopathological observation and the indexes of liver function, such as alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP), after another 4 weeks. Moreover, fasting blood glucose (FBG), and serum levels of total cholesterol (TC), triglyceride (TG) and insulin were measured, and the HOMA insulin resistance index (HOMA-IR) was calculated. The mRNA and protein expression levels of PPARs and inflammation-related factors were measured by qPCR and Western blot, respectively. RESULTS: After treatment with STZ for 7 d, the FBG of mice exceeded 11.1 mmol/L, suggesting that the diabetic model was established. After 4 weeks, the structural deformation of the hepatocytes (including hepatocytes containing abundant fat vacuoles, and inflammatory cell infiltration), and the increases in the serum levels of insulin, HOMA-IR, TC, TG, ALT, AST and ALP were observed (P<0.01), indicating the occurrence and progression of hepatopathy in diabetic mice. Meanwhile, compared with the control group, the mRNA and protein expression of PPARα, PPARβ and PPARγ decreased, but the expression of nuclear factor-κB (NF-κB), cyclooxygenase 2 (COX-2) and inducible nitric oxide synthase (iNOS) significantly increased in the diabetic hepatopathy mice (P<0.01). CONCLUSION: Down-regulation of PPARα, PPARβ and PPARγ and activation of NF-κB-COX-2/iNOS signaling pathways may be involved in the diabetic hepatopathy in mice induced by long-term high-energy diet feeding combined with intraperitoneal injection of STZ.     

Correlation of serum omentin-1 level and insulin resistance in rats

AIM：To establish the insulin resistance rat model for evaluating the correlation of omentin-1 level and insulin resistance. METHODS：SPF male Wistar rats (n=30) were randomly divided into normal control group (NC, n=15) and high-fat diet group (HF, n=15). The rats in NC group were fed with basic diet. The insulin resistant model was established by feeding the rats with high-fat diet in HF group. After 10 weeks, 5 rats in each group were assessed by the technique of hyperinsulinaemic-euglycaemic clamp. After the insulin resistant model was successfully established, the body weight and fasting blood glucose were detected. The concentration of fasting serum omentin-1 was analyzed by ELISA. Fasting serum insulin was measured by radioimmunoassay. RESULTS：No difference of fasting blood glucose between the 2 groups was observed. The level of fasting serum insulin in HF group was significantly higher than that in NC group (P<0.05). The level of serum omentin-1 in HF group were significantly decreased compared with NC group (P<0.01). Pearson’s correlation analysis showed that negative correlations between serum omentin-1 and fasting serum insulin (r=-0.654,P<0.01), serum omentin-1 and free fatty acid (r=-0.446, P<0.05) was found. CONCLUSION：In rats, serum omentin-1 level began to decrease at insulin resistance stage. As serum omentin-1 level decreased, the basal insulin level increased, indicating that decreased serum omentin-1 level may be an early factor of IR, diabetes and cardiovascular diseases.     

Application of PCR-HRM in the study of relationship between leptin gene promoter polymorphisms and liver cirrhosis

AIM: To analyze the feasibility of PCR-high-resolution melting curve analysis (HRM) for detecting the site mutations of C2549 and G2548 in leptin gene promoter from the patients with liver cirrhosis, and to explore the relevance between mutant genotypes and physiological and biochemical indexes in liver cirrhosis patients. METHODS: Compared with the method of PCR-restriction fragment length polymorphism (RFLP), the present research used the method of PCR-HRM to analyze the site mutations of C2549 and G2548 in leptin gene promoter in control group (n=100) and liver cirrhosis group (n=100). The physiological and biochemical indexes of the patients were also detected and compared. RESULTS: Leptin gene promoter polymorphism was detected using PCR-HRM with effectiveness, high flux and accuracy. Preliminary results showed that the main mutation of the patients with liver cirrhosis was in C2549 site, but not found in G2548 site. Leptin, free leptin index (FLI), fasting insulin (FINS) and insulin resistance index estimated by homeostatic model assessment (HOMA-IR) in liver cirrhosis group were higher than those in control group. Insulin sensitivity index (ISI) and soluble leptin receptor (sOB-R) in liver cirrhosis group were lower than those in control group with significant difference except leptin level. Meanwhile, FLI showed positively correlated with FINS and HOMA-IR (r=0.45, r=0.53, P<0.05), and negatively with ISI (r=-0.34, P<0.05). In the patients with liver cirrhosis, C2549A heterozygous mutation was predominant. The indexes of HOMI-IR, leptin, sOB-R and FLI of C2549A homozygotes and heterozygotes were higher than those of the wildtypes, which showed significant difference except leptin and sOB-R levels (P<0.05). CONCLUSION: PCR-HRM can be more accurate for identifying leptin promoter polymorphism. The increase in the frequency of C2549A mutation may be closely related with liver cirrhosis. Existence of hyperinsulinemia and insulin resistance may be correlated with leptin level in the patients with liver cirrhosis.     

Association of serum soluble E-selectin concentrations with insulin resistance in essential hypertension patients

AIM: To investigate the relationship between serum soluble E-selectin (sE-selectin) and insulin resistance, serum uric acid, serum lipid in essential hypertension patients. METHODS: Fasting serum sE-selectin concentration, plasma glucose, serum insulin, serum uric acid, total cholesterol, triglycerides, high density lipoprotein-cholesterol, low density lipoprotein-cholesterol were determined in 186 patients with essential hypertension (75 males, 111 females). Homeostasis model assessment was applied to assess the status of insulin resistance (HOMA-IR). RESULTS: Based on the HOMA-IR, the essential hypertension patients were divided into insulin-sensitive individuals (IS) and insulin resistant subjects (IR). The serum sE-selectin concentration was significantly higher in male group [(50.1±17.8)g/L]than in female group [(40.6±16.6)g/L](P<0.01). No difference between IR group [(51.6±16.8)g/L]and IS group [(48.5±18.8)g/L] in male, and significantly higher in IR group [(45.1±18.0)g/L]than in IS group [(36.0±13.7)g/L](P<0.01) in female were observed. A stepwise multiple linear regression analysis showed that HOMA-IR was an independent predictor of serum sE-selectin concentrations in female group and not in male group, and both serum uric acid and serum lipid were not independent predictors of serum sE-selectin concentrations. CONCLUSION: Serum sE-selectin concentrations were directly related to insulin resistance in females with essential hypertension and not in males with essential hypertension. Both serum uric acid and serum lipid were not directly related to serum sE-selectin concentrations.