辛伐他汀改善肥胖合并骨质疏松雌性大鼠肝脏损伤的作用及机制
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摘要
目的观察辛伐他汀干预对肥胖合并骨质疏松雌性大鼠肝脏损伤的作用,并探讨其可能的机制。方法 40只雌性SD大鼠随机分为假手术对照组(SHAM组)、去卵巢组(OVX组)、去卵巢+高脂组(OVX+HFD组)、去卵巢+高脂+辛伐他汀治疗组(OVX+HFD+SIM组),高脂饲料和辛伐他汀[10 mg/(kg·d)]灌胃分别为16周和8周,葡萄糖和胰岛素耐量试验分析大鼠的胰岛素抵抗情况,检测血脂指标,HE染色观察肝脏的病理学变化,RT-PCR、免疫组化和Western blot分析内质网应激分子ATF6、GRP78、CHOP、NF-κB、CD68、TLR4的表达。结果与SHAM组相比,OVX组和OVX+HFD组大鼠血清三酰甘油(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)水平显著升高(P<0.05),高密度脂蛋白胆固醇(HDL-C)水平明显降低(P<0.05),肝细胞出现水肿变性,TLR4、NF-κB、CD68、GRP78、CHOP的表达显著升高,以OVX+HFD组变化更为明显。与OVX+HFD组相比,OVX+HFD+SIM组大鼠血清TG、TC、LDL-C降低,HDL-C升高,肝细胞的水肿变性明显减少,TLR4及CD68、GRP78、CHOP的表达显著降低。结论辛伐他汀具有显著的降脂和改善胰岛素抵抗的作用,可能是通过减轻肝细胞中内质网应激、炎性细胞浸润来实现的。
Objective To observe the effect of simvastatin treatment on liver damage in obese female SD rats merged with osteoporosis and probe into possible mechanisms. Methods Forty female SD rats were randomly divided into sham operated group( Group Sham),ovariectomized group( Group OVX),ovariectomized group treated with high-fat diet( Group OVX + HFD) and OVX + HFD treated with simvastatin group( Group OVX + HFD + SIM). Glucose tolerance test( GTT) and insulin tolerance test( ITT) were used to examine changes of insulin sensitivity. Serum biochemical indices were detected. HE staining was used to observe the pathological changes in liver tissues. Expression of CD68,TLR4,NFκB,glucose regulated protein 78( GRP78) and GADD153( CHOP) were analyzed by immunohistochemistry,RT-PCR and western blot,respectively. Results Compared with Group SHAM,serum TG,TC and LDL-C levels were significantly higher,while HDL-C level was markedly lower,the structure showed obvious edema and degeneration in liver cells were observed in Group OVX + HFD and Group OVX. Expression of TLR4,CD68,NFκB,GRP78 and CHOP were significantly increased in Group OVX + HFD and Group OVX,the changes were more obvious in Group OVX + HFD.Compared with Group OVX + HFD,serum TG,TC and LDL-C levels were reduced and HDL-C level was increased,while edema and degeneration in liver cells were significantly reduced in Group OVX + HFD + SIM,while expression of TLR4,CD68,NFκB,GRP78 and CHOP were obviously reduced in liver tissue. Conclusion Simvastatin has hypolipidemic effect and alleviate IR,which maybe mediated by the reduction of ERS and inflammatory infiltration in liver tissues.
Objective To observe the effect of simvastatin treatment on liver damage in obese female SD rats merged with osteoporosis and probe into possible mechanisms. Methods Forty female SD rats were randomly divided into sham operated group( Group Sham),ovariectomized group( Group OVX),ovariectomized group treated with high-fat diet( Group OVX + HFD) and OVX + HFD treated with simvastatin group( Group OVX + HFD + SIM). Glucose tolerance test( GTT) and insulin tolerance test( ITT) were used to examine changes of insulin sensitivity. Serum biochemical indices were detected. HE staining was used to observe the pathological changes in liver tissues. Expression of CD68,TLR4,NFκB,glucose regulated protein 78( GRP78) and GADD153( CHOP) were analyzed by immunohistochemistry,RT-PCR and western blot,respectively. Results Compared with Group SHAM,serum TG,TC and LDL-C levels were significantly higher,while HDL-C level was markedly lower,the structure showed obvious edema and degeneration in liver cells were observed in Group OVX + HFD and Group OVX. Expression of TLR4,CD68,NFκB,GRP78 and CHOP were significantly increased in Group OVX + HFD and Group OVX,the changes were more obvious in Group OVX + HFD.Compared with Group OVX + HFD,serum TG,TC and LDL-C levels were reduced and HDL-C level was increased,while edema and degeneration in liver cells were significantly reduced in Group OVX + HFD + SIM,while expression of TLR4,CD68,NFκB,GRP78 and CHOP were obviously reduced in liver tissue. Conclusion Simvastatin has hypolipidemic effect and alleviate IR,which maybe mediated by the reduction of ERS and inflammatory infiltration in liver tissues.
引文
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[17]王丽红,袁林栋,姜雪,等.辛伐他汀对慢性阻塞性肺疾病大鼠外周血和肺泡灌洗液中IL-17、IL-21及RORγt的影响[J].广东医学,2016,37(13):1921-1924.
[18]黄文新,林化,谭宁,等.辛伐他汀对自发性高血压大鼠外周血单个核细胞白细胞介素-1β表达的影响[J].广东医学,2010,31(14):1776-1778.
[2]Hermoso DA,Shimada LB,Gilglioni EH,et al.Melatonin protects female rats against steatosis and liver oxidative stress induced by oestrogen deficiency[J].Life Sci,2016(157):178-186.
[3]Chedraui P,Pérez-López FR,Escobar GS,et al.Circulating leptin,resistin,adiponectin,visfatin,adipsin and ghrelin levels and insulin resistance in postmenopausal women with and without the metabolic syndrome[J].Maturitas,2014,79(1):86-90.
[4]Bj9rnsson ES.Hepatotoxicity of statins and other lipid-lowering agents[J].Liver Int,2017,37(2):173-178.
[5]Wakatsuki A,Okatani Y,Fukaya T.Statin attenuates increase in C-reactive protein during estrogen replacement therapy in postmenopausal women[J].Circulation,2002,106(24):198-199.
[6]Yin H,Shi ZG,Yu YS,et al.Protection against osteoporosis by statins is linked to a reduction of oxidative stress and restoration of nitric oxide formation in aged and ovariectomized rats[J].Eur J Pharmacol,2012,674(2-3):200-206.
[7]Breder I,Coope A,Arruda AP,et al.Reduction of endoplasmic reticulum stress——a novel mechanism of action of statins in the protection against atherosclerosis[J].Atherosclerosis,2010,212(1):30-31.
[8]Jadhav SB,Jain GK.Statins and osteoporosis:new role for old drugs[J].J Pharm Pharmacol,2006,58(1):3-18.
[9]Yida Z,Imam MU,Ismail M,et al.N-Acetylneuraminic acid attenuates hypercoagulation on high fat diet-induced hyperlipidemic rats[J].Food Nutr Res,2015,59:29046.
[10]de Oliveira MC,Gilglioni EH,de Boer BA,et al.Bile acid receptor agonists INT747 and INT777 decrease oestrogen deficiency-related postmenopausal obesity and hepatic steatosis in mice[J].Biochim Biophys Acta,2016,1862(11):2054-2062.
[11]Guarino M,Loperto I,Camera S,et al.Osteoporosis across chronic liver disease[J].Osteoporos Int,2016,27(6):1967-1977.
[12]Cnop M,Foufelle F,Velloso LA.Endoplasmic reticulum stress,obesity and diabetes[J].Trends Mol Med,2012,18(1):59-68.
[13]Hummasti S,Hotamisligil GS.Endoplasmic reticulum stress and inflammation in obesity and diabetes[J].Circ Res,2010,107(5):579-591.
[14]Hosoi T,Ozawa K.Endoplasmic reticulum stress in disease:mechanisms and therapeutic opportunities[J].Clin Sci,2009,118(1):19-29.
[15]Kojanian H,Szafran-Swietlik A,Onstead-Haas LM,et al.Statins Prevent Dextrose-Induced Endoplasmic Reticulum Stress and Oxidative Stress in Endothelial and Hep G2 Cells[J].Am J Ther,2016,23(6):1456-1463.
[16]Kim Y,Lee EJ,Jang HK,et al.Statin pretreatment inhibits the lipopolysaccharide-induced epithelial-mesenchymal transition via the downregulation of toll-like receptor 4 and nuclear factor-κB in human biliary epithelial cells[J].J Gastroenterol Hepatol,2016,31(6):1220-1228.
[17]王丽红,袁林栋,姜雪,等.辛伐他汀对慢性阻塞性肺疾病大鼠外周血和肺泡灌洗液中IL-17、IL-21及RORγt的影响[J].广东医学,2016,37(13):1921-1924.
[18]黄文新,林化,谭宁,等.辛伐他汀对自发性高血压大鼠外周血单个核细胞白细胞介素-1β表达的影响[J].广东医学,2010,31(14):1776-1778.