Effect of YIGU capsule on IGF-I mRNA and protein expression in rat osteoblasts in vitro

AIM: The effects of YIGU capsule on proliferation and IGF-I mRNA protein expressions in osteoblasts were studied. METHODS: (1) Forty 12-month old Sprague-Dawley female rats were divided randomly into four groups (YIGU capsule high dose group, medium dose group and low dose group; saline group), the drug-containing serum and control serum were prepared. (2) The new-born Sprague-Dawley rat osteoblasts were cultured with different YIGU capsule drug-containing serum at different concentrations and different exposure time. MTT method was used to observe proliferation of osteoblasts. (3) RT-PCR method was used to measure the relative IGF-I mRNA levels and ELISA method was used to measure IGF-I secretion at different exposure time. (4) ELISA method was used to measure IGF-I secretion at different exposure time. RESULTS: (1) Proliferation of osteoblasts was more than the control groups after 48, 72 and 96 h, respectively (P<0.01); (2) The relative IGF-I mRNA levels and IGF-I protein expression were higher than those in control group after 48, 72 and 96 h, respectively (P<0.01 or P<0.05). CONCLUSIONS: It was suggested that YIGU capsule drug-containing serum promoted proliferation, IGF-I mRNA and protein expression. These results may be parts of the mechanisms of YIGU capsule to prevent and treat osteoporosis.     

Effect of yigu capsule drug-containing serum on proliferation,ALP activity and expression of interleukin-11 mRNA in rat osteoblasts in the co- culture

AIM:To investigate the effect of yigu caps ule drug-containing serum on proliferation,ALP activity and expression of inter leukin-11 mRNA in Sprague-Dawley rat osteobalsts in the co-culture system.METHODS:(1) Osteoblasts and osteoclasts were isolated from 1 an d 5-day-old Sprague-Dawley rats,respectively.The osteoblasts-osteoclasts co- culture system was built to prevent the two kinds of cells from contact and allo w the media to exchange.The experiment included two groups,drug-containing se ra group and control group.(2) The osteoblasts proliferation,ALP activity and expression of interleukin-11 mRNA were detected by the MTT,4-aminoantipyrine sp ectrometric methods and FQ-PCR，respectively.RESULTS:In drug-containing sera group,the osteoblasts prolifer ation,ALP activity,expression of IL-11 mRNA were higher than those of control group (P<0.05).CONCLUSION:Rat sera containing yigu capsule can obviously enhan ce the osteoblast proliferation,ALP activity and expression of IL-11 mRNA.     

Effect of Yiqi Huayu Huatan decoction on expression of KLF15, HMGB1, NF-κB and its downstream inflammatory factors in rats with UUO-induced renal interstitial fibrosis

AIM: To observe the effect of Yiqi Huayu Huatan decoction (YHHD) on unilaterral ureteral obstruction (UUO)-induced renal interstitial fibrosis in rats, and to investigate the possible mechanism. METHODS: Female SD rats (n=48) were randomly divided into sham group, model group, telmisartan group, and low-, middle-and high-dose YHHD groups, with 8 rats in each group. The UUO model rats was established by ligating left ureter. The rats in sham group and model group were treated with equal volume of normal saline, others were treated with the corresponding drugs daily. After 12 weeks, the rats were sacrificed. The serum samples were collected for determining the concentrations of cystatin C (Cys-C) and uric acid (UA). The morphological changes of the renal tissue were observed by PAS staining. The collagen fiber was observed by Masson staining. The mRNA expression of Krüppel-like factor 15 (KLF15), high-mo-bility group box protein 1 (HMGB1), nuclear factor-κB (NF-κB), IκB, monocyte chemotactic protein-1 (MCP-1), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), fibronectin (FN), collagen type I (Col I) and Col-Ⅳ was detected by real-time PCR. The protein expression of KLF15, HMGB1 and NF-κB was detected by Western blot. The protein expression of MCP-1 was determined by the method of immunohistochemistry. RESULTS: Compared with sham group, the deposition rate of collagen fibers and the concentration of Cys-C in model group were significantly increased (P<0.05), the mRNA and protein expression of KLF15 was significantly down-regulated (P<0.05), while the mRNA expression of HMGB1, NF-κB, IκB, MCP-1, IL-1β, TNF-α, FN, Col I and Col Ⅳ and the protein expression of HMGB1, NF-κB and MCP-1 were significantly up-regulated (P<0.05). Compared with model group, the deposition rates of collagen fibers in middle-and high-dose YHHD groups and telmisartan group were significantly decreased (P<0.05), with down-regulated protein expression of HMGB1 and NF-κB and mRNA expression of IL-1β and TNF-α (P<0.05). The protein expression of KLF15 was significantly up-regulated in high-dose YHHD group and telmisartan group (P<0.05), while the protein expression of MCP-1 and the mRNA expression of FN were significantly down-regulated (P<0.05). The mRNA expression of KLF15 was significantly up-regulated in high-dose YHHD group (P<0.05), while the mRNA expression of MCP-1, Col I and Col IV was significantly down-regulated (P<0.05). The mRNA expression of NF-κB and IκB was significantly down-regulated and the concentration of Cys-C was significantly decreased in each dose of YHHD groups and telmisartan group (P<0.05). No significant difference of UA level among the groups was observed. CONCLUSION: YHHD alleviates renal interstitial fibrosis in a dose-dependent manner, and YHHD at high dose shows the most obvious effect. The mechanism may be associated with the up-regulation of KLF15 and the down-regulation of HMGB1, NF-κB and its downstream inflammation-related factors in the renal tissue.     

Effects of Nrf2 overexpression on proliferation,activation and collagen type I synthesis in cultured hepatic stellate cells stimulated by ethanol

AIM：To investigate the effects of nuclear factor E2-related factor 2 (Nrf2) overexpression on the proliferation, cell cycle distribution, collagen type I (Col I) synthesis and alpha-smooth muscle actin (α-SMA) expression in cultured hepatic stellate cell line HSC-T6 stimulated by ethanol. METHODS：Cultured HSC-T6 cells were transfected with pEGFP-Nrf2 or pEGFP-N1 (empty vector) plasmid by liposome transient transfection. The cells were divided into control group, ethanol group, ethanol+pEGFP-Nrf2 group and ethanol+pEGFP-N1 group. The mRNA expression of Nrf2, α-SMA and Col I was determined by RT-PCR, and their protein expression was detected by Western blotting. Cell proliferation was assessed by MTT assay, and cell cycle was detected by flow cytometry. RESULTS：The pEGFP-Nrf2 plasmid was successfully transfected into HSC-T6 cells, and the mRNA and protein expression of Nrf2 was higher than other three groups 48 h after transfection (P<0.05). Compared with control group, the cell proliferation and the mRNA and protein expression of α-SMA and Col I in ethanol group and ethanol+pEGFP-N1 group were significantly increased (P<0.05), and the numbers of HSC-T6 cells were decreased in G1 phase and increased in S phase (P<0.05), without significant differences between the two groups (P>0.05). Meanwhile, the cells in ethanol+pEGFP-Nrf2 group showed significantly decreased proliferation level, down-regulated mRNA and protein expression of α-SMA and Col I, higher numbers in G1 phase and lower numbers in S phase compared with ethanol group and ethanol+pEGFP-N1 group (P<0.05). CONCLUSION：Nrf2 overexpression could significantly down-regulate the expression of α-SMA and Col I and cause G1/S phase arrest in HSC-T6 cells cultured with ethanol, thus inhibiting the proliferation and activation of the cells.     

Effects of arsenic trioxide on T-bet/GATA3 signal pathway in MRL/lpr mice

AIM: To investigate the effect of arsenic trioxide (ATO) on T-bet/GATA3 signal pathway in MRL/lpr mice.METHODS: MRL/lpr mice and C57BL/6J mice at the age of 20 weeks were chosen and then divided in 2 different sub-groups, respectively. The mice in 2 sub-groups received ATO (0.4 mg·kg^-1·d^-1) and sodium chloride (NS, volume weight-determined) by intraperitoneal injection respectively for 2 months. Afterward, the spleens were isolated from the MRL/lpr and C57BL/6J mice under pathogen-free condition and the suspensions were prepared. The mRNA level of T-bet, GATA3, IFN-γ,IL-4 and the mRNA ratio of T-bet/GATA3 were detected by RT-qPCR. The protein expression of T-bet and GATA3 was determined by Western blot. The serum levels of IFN-γ and IL-4 were measured by ELISA.RESULTS: The mRNA and protein levels of T-bet, IFN-γ and the mRNA ratio of T-bet/GATA3 in NS group of MRL/lpr mice were higher than those in NS group of C57BL/6J mice (P<0.05). However, the GATA3 and IL-4 were lower in NS group of MRL/lpr mice in both mRNA and protein level (P<0.05). In MRL/lpr mice, the mRNA and protein levels of T-bet, IFN-γ and the mRNA ratio of T-bet/GATA3 were lower in ATO group compared with NS group (P<0.05), no difference was found in GATA3 and IL-4. No difference of the indexes mentioned above between ATO group and NS group in C57BL/6J mice was observed.CONCLUSION: ATO may affect the signaling pathway of T-bet/GATA3 to down-regulate the mRNA expression and the protein secretion of IFN-γ by decreasing the expression of T-bet in MRL/lpr mice.     

Effect of combination of Penthorum chinense Pursh and Puerariae flos on L-02 liver cell injury induced by alcohol

AIM: To explore the effect of Penthorum chinense Pursh and Puerariae flos-containing serum on L-02 liver cell injury induced by alcohol and its possible mechanism. METHODS: After preparing drug-containing serum, the L-02 cells cultured in vitro were divided into 6 groups:blank control group, model group, 1:1 group, 2:1 group and 1:2 group of combination of Penthorum chinense Pursh and Puerariae flos, and tiopronin group. The viability of the L-02 cells was measured by MTT assay. The activity of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and superoxide dismutase (SOD), and the content of malondialdehyde (MDA) were detected by enzyme label methods. The expression of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) at mRNA and protein levels was determined by real-time PCR and Western blot, respectively. RESULTS: Compared with control group, the levels of ALT, AST and MDA were increased significantly, and SOD was decreased in model group (P<0.01). Compared with model group, these indexes in all treatment groups were opposite (P<0.01). Compared with control group, the expression of TNF-α and IL-6 at mRNA and protein levels was significantly increased, the mRNA and protein expression of Nrf2 and HO-1 was decreased (P<0.01). Compared with model group, these indexes in combination groups were opposite (P<0.01). CONCLUSION: The therapeutic effects of Pentehorum chinensa Pursh and Puerariae flos-containing serum may affect the expression levels of TNF-α, IL-6, Nrf2 and HO-1, and reduce the inflammatory reaction and oxidative stress in alcohol-induced L-02 liver cells, which plays a role in attenuating alcoholic liver injury.     

Effects of renal denervation on inflammatory factors in a rabbit model of early atherosclerosis

AIM: To evaluate the effects of renal denervation (RDN) on the expression of tumor necrosis factor-α (TNF-α), interleukin-1α (IL-1α) and interleukin-6 (IL-6) in a rabbit model of early atherosclerosis. METHODS: New Zealand male rabbits were divided into control group, RDN+ high-fat diet (HFD) group (RDN group), sham+HFD group (sham group) and HFD group. The rabbits in later 3 groups were fed with 2% cholesterol for 8 weeks to establish an early atherosclerosis model. The blood samples were collected to test the levels of lipids, norepinephrine (NE), TNF-α, IL-1α and IL-6. The protein expression of angiotensin Ⅱ (Ang Ⅱ) was detected by the method of immunohistochemistry. The levels of nuclear factor-κB (NF-κB) and Ang II 1 type receptor (AT1R) were evaluated by Western blot. The mRNA expression of TNF-α, IL-1α and IL-6 was determined by real-time PCR. RESULTS: After 1 d of RDN procedure, the NE level was lower in RDN group than that in sham group (P<0.01). After 8 weeks, the NE level was lower in RDN group than that in sham group and HFD group (P<0.05), and triglyceride (TG) was lower in RDN group than that in HFD group (P<0.05). The protein expression of Ang II was decreased in RDN group compared with sham group and HFD group (P<0.01). The protein expression of NF-κB was lower in RDN group than that in sham group (P<0.05). The plasma levels of TNF-α and IL-1α were reduced in RDN group compared with sham group and HFD group (P<0.05). The mRNA expression of TNF-α, IL-1α and IL-6 was reduced in RDN group compared with sham group (P<0.05). CONCLUSION: RDN inhibits sympathetic activity, decreases the plasma level of TG, and alleviates inflammatory reactions in the rabbits with atherosclerosis.     

Palmitate triggers inflammatory response by upregulating fatty acid translocase in THP-1 macrophages

AIM: To investigate the role of fatty acid translocase (FAT/CD36) on palmitate-induced inflammation in human monocyte-derived macrophage THP-1.METHODS: THP-1 cells were treated with palmitate (0, 0.1 and 0.2 mmol/L) for 24 h. Transwell chamber assay was used to examine the migration ability of THP-1 cells. The mRNA expression of CD36, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and monocyte chemotactic protein 1 (MCP-1) was measured by real-time PCR. The protein levels of TNF-α and IL-6 in the supernatant of cultured cells were measured by ELISA. The protein level of CD36 was examined by Western blot. Small interfering RNA (siRNA) targeting CD36 (siCD36) was used to inhibit the expression of CD36 in the THP-1 cells, and the changes of the cell migration and inflammatory response were monitored as mentioned above. RESULTS: Palmitate increased the expression of CD36 in the THP-1 cells (P<0.05). Palmitate also up-regulated inflammatory cytokine and chemokine levels, and the differences were statistically significant (P<0.05). Compared with control group, palmitate promoted migration of THP-1 cells. siCD36 was transfected into the THP-1 cells and the silencing efficiency was approximately 54%. The protein levels of TNF-α and IL-6 were also decreased in siCD36 group compared with scrambled RNA (scrRNA) group, and the differences were statistically significant (P<0.05). The migrated cells in siCD36 group were significantly less than those in scrRNA group (P<0.05).CONCLUSION: Palmitate promotes migration ability and triggers inflammatory response in the THP-1 macrophages by upregulating CD36 expression.     

Preliminary study of IL-17 signal transduction pathway in peripheral blood mononuclear cells from lupus nephritis patients

AIM: To investigate the role of IL-17 and its signal conduction component-JNK activity in the pathogenesis of LN. METHODS: Peripheral blood mononuclear cells (PBMC) were separated and cultured from 15 cases of active lupus nephritis (LN) patients. IL-6 level was detected by ELISA, IL-6 mRNA was checked with RT-PCR, and JNK activity was measured by Western blot. RESULTS: At same IL-17 end concentrations, there was a much higher level of IL-6 in LN group than in control group (all P<0.05). IL-17 induced a significant elevation of IL-6 mRNA expression and JNK activity in PBMC from LN patients in a time- and dose-dependent manner, which could be blocked markedly by IL-17 monoclonal antibody, mIgG₂₈, and dexamethasone. Much higher IL-6 mRNA expression was observed in LN group than in control group under medium culture or IL-17-conditioned culture (all P<0.01). Under medium culture or IL-17-conditioned culture, there was much higher JNK activity of PBMC in active LN group than that in control group. There was a positive linear correlation between PBMC JNK activity and their SLEDAI or IL-6 mRNA expression level in LN patients (r1=0.638, P<0.01; r2=0.644, P<0.01). CONCLUSION: These results suggest that IL-17 may take part in the initiation and progression of LN through induction of IL-6 overexpression by PBMC,and JNK hyperactivity may be necessary in the IL-17 signal transduction.     

Effect of fibrin on expression of interleukin-6 in rat brain vascular endothelial cells

AIM:To investigate the kinetics of interleukin-6 (IL-6) protein and mRNA expressions in the co-culture of rat brain vascular endothelial cells(VECs) with fibrin. METHODS:The IL-6 concentration were measured by rat IL-6 ELISA kit and the IL-6 mRNA was measured by real-time PCR after development an in vitro model of in situ fibrin polymerization on rat brain vascular endothelial cells. RESULTS:Fibrin induced IL-6 expression. The IL-6 antigen concentration increased significantly in 1.0 g/L media group compared to the low dose fibrin group or control media group (P<0.01). Fibrin-induced IL-6 expression in rat brain VECs as early as 2 h post-fibrin stimulation, increased significantly at 8 h, keeping at high level till 24 h (P<0.05). Collagen did not induce IL-6 expression at 24 h. Real-time PCR analysis showed that IL-6 antigen increase in fibrin treated rat brain VECs was due to fibrin induced elevation of steady state mRNA expression. CONCLUSION:Fibrin is a potent vascular endothelial cell activator. It induces IL-6 expression in time and dose dependent fashions.     

Effects of CARHSP1 gene expression on viability,apoptosis and immune factors of vascular endothe-lial cells induced by hypoxia in atherosclerosis

AIM: To investigate the effect of calcium-regulated heat stable protein 1 (CARHSP1) gene expression on the viability, apoptosis and expression of interleukin-6 (IL-6) and C-reactive protein (CRP) in vascular endothe-lial cells induced by hypoxia.METHODS: The protein expression of CARHSP1 was detected by Western blot in atherosclerotic plaques. Human umbilical vein endothelial cells (HUVECs) were treated with hypoxia, and the cells were divided into normal culture group, hypoxia group, hypoxia+CARHSP1-siRNA group and hypoxia+pcDNA3.1-CARHSP1 group. The viability and apoptotic rate of the HUVECs were measured by CCK-8 assay and flow cytometry, respectively. The mRNA expression of IL-6 and CRP was detected by RT-PCR. The protein levels of caspase-3, cleaved caspase-3, Bcl-2 and Bax were determined by Western blot.RESULTS: The protein expression of CARHSP1 in atherosclerotic plaques was significantly higher than that in control group (P<0.05). Hypoxia significantly increased the expression of CARHSP1. The cell viability and the protein expression of Bcl-2 were significantly lower in hypoxia group than those in normal culture group (P<0.05). The apoptotic rate and the protein levels of IL-6, CRP, cleaved caspase-3 and Bax were significantly higher than those in normal culture group (P<0.05). Compared with hypoxia group, the cell viability and protein expression of Bcl-2 were significantly increased in hypoxia+CARHSP1-siRNA group, while the apoptotic rate and the protein levels of IL-6, CRP, cleaved caspase-3 and Bax were decreased significantly (P<0.05). The cell viability and protein expression of Bcl-2 were decreased significantly in hypoxia+pcDNA3.1-CARHSP1 group, while the apoptotic rate and the protein le-vels of IL-6, CRP, cleaved caspase-3 and Bax were increased significantly (P<0.05).CONCLUSION: The expression of CARHSP1 is increased in atherosclerotic plaques, and inhibition of CARHSP1 expression improves the viability, reduces the apoptosis, and down-regulates the expression of IL-6 and CRP in the HUVECs. Over-expression of CARHSP1 exerts the opposite effect.     

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.     

The effect of postconditioning on apoptosis-related gene expression of lung tissues in rat intestinal ischemia-reperfusion

AIM: To investigate the effect of postconditioning on apoptosis-related gene expression of lung tissues in rat intestinal ischemia-reperfusion(I/R). METHODS: Male Sprague-Dawley rats were divided into 3 groups which were sham operation group (sham), I/R group and ischemic postconditioning (IPOST) group. The experimental animals were sacrificed after reperfusion for 6 h. The tissue sections were stained with hematoxylin and eosin and examined under light microscope to observe lung histopathological changes. Lung W/D ratios were measured. Lung tissue samples were taken to detect Bcl-2, Bax and caspase-3 mRNA and protein expression using RT-PCR and Western blotting analysis, respectively. RESULTS: Compared with I/R group, the morphological changes of lung were alleviated and the lung W/D ratios significantly decreased in IPOST group (P<0.05). The mRNA and protein expression of Bcl-2 in IPOST group was obviously higher than that in I/R group (P<0.05), while the mRNA and protein expression of Bax and caspase-3 in IPOST group was obviously higher than that in I/R group (P<0.05). CONCLUSION: Ischemic postconditioning attenuated intestinal ischemia-reperfusion-induced acute lung injury partly by increasing Bcl-2 mRNA and protein expression, reducing Bax mRNA and protein expression.     

Effects of high glucose and homocysteine on expression of MMP-9 in rat fibroblasts

AIM: To observe the effect of different concentrations of glucose and homocysteine on the expression of MMP-9 in fibroblasts, and to investigate the relationship between homocysteine and diabetic foot.METHODS: Fibroblasts were obtained from the dermis of 1-day-old normal SD rat and cultured in DMEM containing 10% FBS. After deprivation of serum (using 0.5% FBS) for 24 h, the fibroblasts in 2-4 passages were cultured for 6 h under the conditions of normal glucose, high glucose and high glucose with high concentration of homocysteine, respectively. The expression of MMP-9 at mRNA and protein levels was assessed by RT-PCR and Western blotting, and the activity of MMP-9 was determined by gelatin zymography analysis. RESULTS: The expression of MMP-9 at mRNA and protein levels and activity of MMP-9 in high glucose (22 mmol/L) group were 1.15 folds, 1.59 folds and 1.34 folds of those in normal glucose group (P<0.05), respectively. The effect of high glucose became more pronounced when co-treated with high concentration of homocysteine (100 μmol/L), which were 1.25 folds, 2.63 folds and 2.52 folds of those in normal glucose group (P<0.05), respectively.CONCLUSION: The increase in the expression of MMP-9 is concentration-dependen with glucose content in the culture. High concentration of homocysteine promotes this process, suggesting that hypercysteinemia might play an important role in the expression of MMP-9 in fibroblast.     

Hypoxia-inducible factor-1α attenuates myocardial inflammatory injury induced by ischemia and reperfusion in rats

AIM:To investigate the role of hypoxia-inducible factor-1α (HIF-1α) stable expression in myocardial inflammatory injury induced by ischemia and reperfusion (I/R) in rats. METHODS:Male Wistar rats were randomly divided into 4 groups:sham operation (sham) group, I/R group, HIF-1α stabilizer dimethyloxalyl glycine (DMOG)+I/R group and HIF-1α inhibitor YC-1+I/R group. The protein expression of myocardial Toll-like receptor 4 (TLR4) and nuclear factor-κB (NF-κB) was determined by Western blot. The mRNA levels of interleukin (IL)-1β, tumor necrosis factor-α (TNF-α), IL-6, TLR4 and NF-κB were detected by real-time PCR. The myeloperoxidase (MPO) activity in the myocardial tissues was measured. HE staining was used to observe the infiltration of inflammatory cells. RESULTS:HIF-1α decreased the infiltration of inflammatory cells, the MPO activity, and the mRNA levels of inflammatory factors IL-1β, IL-6 and TNF-α in the myocardial tissues. HIF-1α also reduced the expression of TLR4 and NF-κB at mRNA and protein levels (P<0.05). CONCLUSION:The stable expression of HIF-1α has an anti-inflammatory effect on the myocardial tissues after I/R injury in rats. The mechanism may be related to the inhibition of TLR4/NF-κB signaling pathway.     

Effects of panax notoginseng saponins on content of triglyceride and the mRNA expression of LXRα in steatotic hepatocyte L02

AIM: To study the effects of panax notoginseng saponins (PNS) on the content of triglyceride (TG) and the mRNA expression of liver X receptor α (LXRα) in steatotic hepatocyte L02.METHODS: The cells of hepatocyte L02 were cultured with 50% bovine calf serum for 48 h and a model of steatosis of hepatocytes was established. The appropriate concentrations of PNS on steatotic hepatocytes were detected by MTT assay. The cells were divided into 5 groups: normal control group, model group, spontaneous recovery group, low concentration of PNS group (10 mg/L) and high concentration of PNS group (50 mg/L). Meanwhile, hepatocytes in model group were continuously cultured by RPMI-1640 medium containing 50% bovine calf serum while others were cultured by RPMI-1640 medium containing 10% bovine calf serum. After 24 h, the lipid droplets in the cells stained with oil red O were observed under microscope, the intracellular TG levels were determined by automatic biochemical analyzer and the mRNA expression of LXRα in hepatocytes was examined by RT-PCR. RESULTS: Compared to normal control group, Oil red O staining presented numerous orange-red or red lipid droplets in the cytoplasm of human L02 hepatocytes in model group. The content of TG in model group were significantly increased (P<0.01). After treated with PNS for 24 h, the content of TG in PNS treatment groups was significant decreased than that in spontaneous recovery group (P<0.05), especially in low concentration of PNS group (P<0.01). The accumulation of lipid droplets in low concentration of PNS group was decreased significantly. Compared to normal control group, the mRNA expression of LXRα in model group was significantly up-regulated (P<0.01). Compared to spontaneous recovery group, the mRNA expression of LXRα in PNS treatment groups declined in different levels, especially in low concentration group (P<0.05).CONCLUSION: The deposition of lipid in hepatocytes might be related to the high expression of LXRα mRNA. PNS decreases the content of TG in steatotic hepatocytes and promotes the recovery of hepatocyte steatosis by downregulating the mRNA level of LXRα.