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Objective:To investigate the mechanism of Shugan Wendan decoction in treating atherosclerosis based on liver X receptor α(LXRα)/nuclear factor-κB(NF-κB) signal. Method:A New Zealand rabbit model of atherosclerosis with liver-Qi stagnation was established by using calf serum albumin immune injury, high fat feeding and bondage emotional stress method. Theses rabbits are randomly divided into 6 groups, control group,model group, atorvastatin group, Shugan Wendan decoction low, medium and high dose group(2.18,6.54,19.62 g·kg-1·d-1). After successful modeling, the rabbits were treated by injecting drugs with Atorvastatin and low, middle and high dose Shugan Wendan decoction to gastric.The control group and the model group were given intragastric administration of saline in the same volume. The period of gavage is 6 weeks. The pathological changes of the rabbits were detected by hematoxylin-eosin(HE) staining.Serum levels of totalcholesterol(TC), triglyceride(TG),low density extremityprotein(LDL-C), high density extremity protein(HDL-C), nitric oxide(NO), and endothelin-1(ET-1) of the rabbits were detected by enzyme method, nitrate reductase method, and enzyme-linked immunosorbent assay(ELISA), respectively.The gene expression of CRP, IL-1β, IL-6 and MMP-9 in the aorta was detected by Real-time fluorescent quantitative polymerase chain reaction(Real-time PCR) method.The protein expression of LXRα/NF-κB signaling pathway wasdetected by Western blot. Result:Compared with normal control group, in model group, the lumen of the blood vessels was significantly narrowed, atheromatous plaques were formed, and a large number of intracellular foam-like changes were seen. In atorvastatin group and Shugan Wendan decoction group, the blood vessels in high, middle, and low concentration groups were narrowed. Atherosclerotic plaques and foam-like changes were all lower than the model group.Compared with the normal control group, the TG, TC, and LDL-C levels in the model groupincreased(PPPPβ, IL-6 and MMP-9 all increased(Pα protein in the model group was decreased(PκB was increased(PPPβ, IL-6 and MMP-9 in the atorvastatin group,the low, middle and high dose Shugan Wendan decoction groups all decreased(Pα protein in the group was increased(PκB was decreased(PConclusion:Shugan Wendan decoction can inhance the function of vascular endothelial cells and the stability of atherosclerotic plaque by regulating LXRα/NF-κB signaling pathway.
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Objective:To observe the expression levels of niemann-pick C1-like 1 (NPC1L1) and adenosine triphosphate-binding cassette transporters G8 (ABCG8) in intestine of hyperlipidemic model rats, in order to investigate the therapeutic mechanism of Shuangyu Tiaozhi decoction on hyperlipidemia. Method:A total of 40 SD rats were selected, including 8 for normal control group. The remaining 32 rats were used to establish hyperlipemic model. After modeling, the rats were randomly divided into the model group (equivalent normal saline), the high and low-dose Shuangyu Tiaozhi groups (15.6, 7.8 g·kg-1), and the Simvastatin group (4 mg·kg-1), with 8 in each group. They were given drugs by gavage for 8 weeks. The levels of total cholesterol (TC), triglyceride (TG) in serum and total cholesterol (TTC), free cholesterol (FTC) in liver of rats in each group were determined by biochemical and enzymatic methods. The morphological changes of liver were observed by hematoxylin-eosin (HE) staining, and the levels of expressions of NPC1L1, ABCG8 and liver X receptor-α (LXR-α) in intestine were detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot. The expression of ABCG8 protein was determined by immunohistochemistry. Result:After successful replication of the hyperlipidemia model, the blood lipid level was abnormally increased, and the liver steatosis became obvious in the model group compared with the normal control group. The expression levels of NPC1L1, LXR-α and ABCG8 increased significantly (PPα were significantly down-regulated, but ABCG8 was obviously up-regulated in a dose-dependent manner (PPPPPPConclusion:Shuangyu Tiaozhi decoction can reduce the blood lipid level of hyperlipemic rat model by reducing the absorption of cholesterol. Its mechanism may be correlated with the down-regulation of NPC1L1 expression and the up-regulation of ABCG8 expression.
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OBJECTIVE:To investigate the effect and mechanism of Allii macrostemonis bulbus on blood lipid levels in hyperlipidemia model rats,and to provide reference for clinical use of Allii macrostemonis bulbus to reduce blood lipid. METHODS:A total of 10 normal rats were included in normal control group and given common diet. Other 50 rats were given hyperlipid diet to induce hyperlipidemia rat model. 40 model rats were randomly divided into model group(hyperlipid diet),Allii macrostemonis bulbus low-dose,medium-dose and high-dose groups(0.83,1.67,2.50 g/kg,fed by hyperlipid diet which containing 10%Allii macrostemonis bulbus 8.3,16.7,25.0 g/kg,fill with hyperlipid feed in patients with insufficient food intake). After fed for 45 d,the contents of TC,TG,LDL-C and HDL-C in serum of rats were detected. Liver,spleen,renal and cardiac indexes of rats were calculated. mRNA expression of low density lipoprotein receptor(LDLR)and liver X-receptor α(LXRα)were detected in liver tissue of rats. RESULTS:Compared with normal control group,the contents of TC and LDL-C in serum and liver index of rats were increased significantly in model group,while the content of HDL-C in serum and mRNA expressions of LDLR and LXR α in liver tissue were decreased significantly,with statistical significance(P<0.01). Compared with model group,the contents of TC and LDL-C in serum were decreased significantly in Allii macrostemonis bulbus groups,while the content of HDL-C was increased significantly. mRNA expressions of LDLR and LXR α in liver tissue were increased significantly in Allii macrostemonis bulbus medium-dose and high-dose groups,while liver and spleen indexes were decreased significantly,with statistical significance(P<0.05 or P<0.01). CONCLUSIONS:Allii macrostemonis bulbus shows good blood-lipid lowering effect,the mechanism of which may be associated with up-regulating mRNA expressions of LDLR and LXRα in liver tissue.
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Objective To observe the lipid regulation role of electroacupuncture intervention in the rats with non-alcoholic fatty liver disease(NAFLD)induced with high fat and cholesteol forage,and to clarify the regulation mechanism of serum retinol binding protein 4 (RBP4 )level and liver X receptor a (LXR-α)and sterol regulatory element binding protein-1c(SREBP-1c)expressions.Methods 44 female SD rats were fed for 7 d to adapt the environment and were randomly divided into normal group, model group, Dongbaogantai group, electroacupuncture group;11 rats in each group.The rats in normal group got routine feeding,and the others were fed with high fat and high cholesterol forage. After 8 weeks, the models were established, the rats in electroacupuncture group were treated with electroacupuncture method (1.5-2.0 Hz, D.-D.wave, 9V, 1-3 mA)in“Ganshu”,“Pishu”,“Geshu”for 15 min,once a day,lasted for 28 d.The changes of fasting blood-glucose(FBG),serum free fatty acids(FFA)and liver tissue homogenate triglyceride(TG)and total cholesterol (TC)levels of the rats in various groups were tested, and enzyme-linked immunosorbent (ELISA)was used to determine the serum RBP4 levels, and Western blotting method was used to detect the LXR-αand SREBP-1 c protein expression levels in rat liver tissue.Results Compared with normal group,the FBG,serum FFA,TG and TC levels in liver tissue homogenate and serum RBP4 level of the rats in model group were increased (P<0.01);the LXR-αand SREBP-1c protein expression levels were also increased (P<0.01).Compared with model group, the FBG,serum FFA,the TG and TC levels in liver tissue homogenate and serum RBP4 levels of the rats in electroacupuncture group and Dongbaogantai group were decreased (P<0.01);the LXR-αand SREBP-1c protein expression levels were also decreased (P< 0.05 or P< 0.01 ). Conclusion Electroacupuncture method in“Fenglong”,“Zusanli”,“Sanyinjiao”can reduce the serum RBP4 level, regulate the lipid metabolism, and improve the lipid deposition of the NAFLD rats;they have obvious therapeutic effect on NAFLD, and its mechanism may be related to inhibiting the increasing of LXR-αand SREBP-1 c protein expressions in liver tissue.
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Objective To investigate the effect of liver X receptor α (LXRα) gene silencing on lipid metabolism-related genes in HepG2. 2. 15 cells. Methods HepG2. 2. 15 cells were divided into blank control group (without transfection), negative control group (transfected with HK plasmid), and shLXRα group(transfected with shLXRα plasmid). The shLXRα plasmids carrying LXRα gene were constructed and were used to transfect HepG2. 2. 15 cells using Polyjet™ reagent. Green fluorescent protein and LXRα protein expression were examined by fluorescence microscope and Western blotting analysis 24-96 h after transfection, so as to identify the best interference time. Then cells were treated with agonis T090131 fo 2 o 4 h, and the content of triglyceride (TG) was observed to detect the degree of steatosis by biochemical assay. The expression of sterol regulatory element binding protein-lc (SREBP-lc) mRNA was detected by RT-PCR and the expression of hepatitis B virus X (HBx) protein and fatty acid synthase (FAS) protein was tested by Western blotting analysis. Results The shLXRa plasmid was constructed and transfected into HepG2. 2. 15 cells successfully. Compared with blank and negative control groups, LXRα protein was markedly decreased in the shLXRα group, with the lowest level found at 48-72 h after transfection (P<0. 01). Afte cell wer stimulate wit T090131, HBx and FAS protein expression, the content of TG, and SREBP-lc mRNA expression gradually increased with the prolongation of stimulation period, and there was no significant difference in HBx expression at the same time point between different groups. FAS protein, TG contents, and SREBP-lc mRNA in shLXRα group were significantly lower than those in the other two groups (P<0. 01). Conclusion HBx can regulate lipid metabolism through LXRa/SREBP-lc/FAS pathway.
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Purpose To examine the regulatory effect of recombinant human fibroblast growth factor-21 on the expression of liver X receptor α and glucose transporter protein 1 in the type 2 diabetes mellitus rats.Methods The rat models of type 2 diabetic mellitus were divided into four groups at random, ic. rhFGF-21 every day, after eight weeks of these treatment, Inspect the fasting blood glucose (FBG), fructosamine(FA), triglyceride(TG), T-cholesterol(TC), high density lipoprotein cholesterol(HDL-C) and low density lipoprotein cholesterol(LDL-C) of these rats, then detecting the mRNA expression of LXRα and GLUT1 by RT-PCR.Results (1) rhFGF-21 can reduce blood glucose steadily to near normal levels in diabetic rats. (2) The expression of LXRα and GLUT1 level was significantly higher in the rhFGF-21 treatment group than that in the model group. (3) rhFGF-21 megadoses and middle doses decreased FA, TG, TC,and LDL-C and elevated HDL-C.Conclusion rhFGF-21 could regulate the mRNA expression of LXRα and GLUT1 in diabetes rats, increase basal level glucose transport, then reduce blood glucose, improve lipid metabolize dysfunction.
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Objective To investigate the effects of berberine on cholesterol efflux in THP-1 macrophage derived foam cells, and explore the possible mechanism. Methods HP-1 cells were induced into macrophages by phorbol myristate acetate (PMA), and were treated with acetylated low-density lipoprotein (Ac-LDL) to establish the THP-1 macrophage derived foam cell models. Foam cells were divided into blank control group and berberine (5 to 20 μmol/L) treatment groups according to the way of treatment and berberine concentrations. After treatment for 24 h, flow cytometry was employed to detect AcLDL aggregation, enzymic method was adopted to detect contents of cholesterol and triglyceride, scintillation counting technique was used to detect cholesterol efflux, and effects of peroxisome proliferator-activated receptor-γ (PPARγ) antagonist GW9662 pretreatment on cholesterol efflux (pioglitazone as positive control) were analysed. Besides, RT-PCR was applied to detect expression of liver X receptor α (LXRα) and ATP binding cassette transporter A1 (ABCA1) mRNA. ResultsCompared with blank control group, AcLDL aggregation and contents of cholesterol and triglyceride of foam cells in various berberine treatment groups decreased significantly (P＜0.01), while cholesterol efflux increased (P＜0.01) in a dose-dependent manner. After GW9662 pretreatment, there was no significant difference in cholesterol efflux between various berberine treatment groups and control group (P>0.05). Furthermore, expression of LXRα and ABCA1 mRNA of foam cells in various berberine treatment groups was higher than that in blank control group. Conclusion Berberine may increase cholesterol efflux in THP-1 macrophage derived foam cells, the mechanism of which may be associated with activation of PPARγ pathway and increase of expression of LXRα and ABCA1 mRNA.