三七对酒精性肝病大鼠肝组织SREBP1c表达的影响

中国临床药理学与治疗学 ›› 2011, Vol. 16 ›› Issue (2): 148-154.

三七对酒精性肝病大鼠肝组织SREBP1c表达的影响

苏艳霞¹, 荀运浩², 陈芝芸³, 施军平²

¹浙江中医药大学,杭州 310053,浙江;
²浙江中医药大学附属杭州第六医院,杭州 310014,浙江;
³浙江中医药大学附属第一医院消化研究室,杭州 310006,浙江

收稿日期:2010-11-10 修回日期:2011-01-12 发布日期:2011-04-20
通讯作者: 施军平,男,主任医师,医学博士,硕士研究生导师,主要从事脂肪性肝病研究。Tel: 0571-85463966 E-mail: davidshi0571@126.com
基金资助:
杭州市医药卫生科技计划项目(2006B056)

Effect of Sanchi on hepatic expression of SREBP1c in rats with alcoholic liver disease

SU Yan-xia¹, XUN Yun-hao², CHEN Zhi-yun³, SHI Jun-ping²

¹Zhejiang University of TCM, Hangzhou 310053, Zhejiang, Chian;
²Affiliated Hangzhou NO.6 Hospital of Zhejiang University of TCM, Hangzhou 310014, Zhejiang, China;
³Digestion Research Room, the NO.1 Hospital of Zhejiang University of TCM, Hangzhou 310006, Zhejiang, China

Received:2010-11-10 Revised:2011-01-12 Published:2011-04-20

摘要/Abstract

摘要： 目的:观察三七对酒精性肝病 (ALD) 大鼠肝组织固醇调节元件结合蛋白(SREBP)1c表达的影响。方法: 110只雄性SD大鼠随机分为正常组30只,模型组35只,三七高、低剂量组和硫普罗宁组各15只,连续14周白酒-玉米油-吡唑混合液灌胃建立ALD模型,同时三七高、低剂量组和硫普罗宁组分别灌服1.2、0.6 g/(kg·d)的三七粉及0.1 g/(kg·d)的硫普罗宁;4周、8周、14周分别处死正常组大鼠10只,模型组8只;14周处死剩余大鼠。光镜观察肝脂肪变、炎症及纤维化程度,RT-PCR法检测肝组织SREBP1c及其下游靶基因乙酰辅酶A合成酶(ACS)、脂肪酸合成酶(FAS) mRNA表达,免疫组化法检测肝组织SREBP1c蛋白表达。结果: (1)各时间点模型组大鼠肝组织脂肪变及炎症程度计分较同期正常组明显增高(P<0.01)。(2)正常组大鼠肝脏可见一定量SREBP1c、ACS、FAS mRNA表达,随时间延长模型组大鼠3个基因的表达均逐步增加,8周、14周时较正常组和模型4周组明显增高(P<0.01或0.05)。(3)模型8周、14周组较同期正常组肝组织SREBP1c表达明显增高(P<0.01或0.05)。(4)三七高、低剂量组及硫普罗宁组大鼠14周肝组织脂肪变及炎症程度、肝脏SREBP1c蛋白以及3个目标基因的表达均较模型组明显降低(P<0.01或0.05)。结论:三七可明显减轻ALD大鼠肝组织脂肪变和炎症程度,抑制肝脏脂代谢关键信号分子SREBP1c mRNA和蛋白及其靶基因ACS、FAS mRNA的表达,从而改善肝组织病理,阻止ALD的慢性化进程。

关键词: 酒精性肝病, 三七, 固醇调节元件结合蛋白-1c

Abstract: AIM: To evaluate the effect of Sanchi on hepatic expression of sterol regulatory element binding protein (SREBP)-1c in rats with alcoholic liver disease(ALD).METHODS: 110 SD male rats were divided into normal group(n=30), control group(n=35), high-dose Sanchi group(n=15), low-dose Sanchi group(n=15) and Tiopronin group(n=15) randomly. A formula of ethanol-corn oil-pyrazole mixture was selected to induce ALD model by feeding for 14 weeks continually. Meanwhile, rats of high-dose Sanchi group, low-dose Sanchi group and Tiopronin group were administered with 1.2 g/(kg·d) Sanchi,0.6 g/(kg·d) Sanchi and 0.1 g/(kg·d) Tiopronin respectively. For investigating dynamic histologic changes,10 rats in normal group and 8 rats in control group were sacrificed at the 4th week,8th week,14th week by turns, and the others were sacrificed at 14th week. Histopathological scores were assessed by microscopic examination of HE and Masson staining of the right lobe of liver. The hepatic expression of SREBP1c were detected with immunohistochemistry, mRNA levels of SREBP1c and the targeting gene, acetyl-CoA-synthetase(ACS) and fatty acid synthetase(FAS) were detected by RT-PCR.RESULTS:(1)Compared with the rats in normal group, rats in control group developed higher steatosis scores and inflammation scores at every point (P<0.01). (2)In normal group, the hepatic expression of SREBP1c, ACS and FAS mRNA were low, which increased significantly as the time flied in model rats (P<0.01 or 0.05). (3)Hepatic expression of SREBP-1c in model group were higher than those of normal group at 8th and 14th week(P<0.01or 0.05), respectively. (4)High-dose Sanchi,low-dose Sanchi,or Tiopronin ameliorated the histopathological changes remarkably(P<0.01 or 0.05), the mRNA of hepatic SREBP-1c, ACS and FAS, together with SREBP-1c, were obviously inhibited(P<0.01 or 0.05).CONCLUSION: Sanchi can remarkably improve the hepatic steatosis and inflammation, restrain the hepatic expression of SREBP1c, a pivotal signal regulating liver lipid metabolism, and the mRNA of hepatic SREBP1c, ACS and FAS, then delay the exacerbation of ALD.

Key words: Liver disease, Alcoholic, Rat, Sanchi, SREBP1c

中图分类号:

R965.1

苏艳霞, 荀运浩, 陈芝芸, 施军平. 三七对酒精性肝病大鼠肝组织SREBP1c表达的影响[J]. 中国临床药理学与治疗学, 2011, 16(2): 148-154.

SU Yan-xia, XUN Yun-hao, CHEN Zhi-yun, SHI Jun-ping. Effect of Sanchi on hepatic expression of SREBP1c in rats with alcoholic liver disease[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2011, 16(2): 148-154.

参考文献

[1] 中华医学会肝病学分会脂肪肝和酒精性肝病学组.酒精性肝病诊疗指南[J].中华肝脏病杂志,2006,14(3):164-166.
[2] 陈芝芸,严茂祥,蔡丹莉,等.三七抗酒精性脂肪肝的实验研究[J].中华中医药杂志, 2006,21(10):614-616.
[3] 刘庆生,王小奇,蔡丹莉,等.三七对酒精性肝病大鼠肝组织NFκB/IκB表达的影响[J].中国药学杂志,2007,42(18):1372-1376.
[4] 蔡丹莉,陈芝芸,严茂祥,等.三七对酒精性脂肪肝大鼠肝组织环氧合酶-2表达的影响[J].中华中医药学刊,2007,25(7):1391-1393.
[5] Horton JD, Goldstein JL, Brown MS. SREBPs:activators of the complete program of cholesterol and fatty acid synthesis in the liver[J].J Clin Invest,2002,109(9):1125-1131.
[6] You M, Fischer M, Deeg MA, et al. Ethanol induces fatty acid synihesis pathways by activation of sterol regulatory element binding protein[J].J Biol Chem,2002,277(32):29342-29347.
[7] Kohjima M,Higuchi N,Kato M,et al.SREBP-1c,regulated by the insulin and AMPK signaling pathways, plays a role in nonalcoholic fatty liver disease[J].Int J Mol Med,2008,21(4):507-511.
[8] Kim KH,Shin HJ,Kim K,et al. Hepatitis B virus X protein induces hepatic steatosis via transcriptional activation of SREBP1 and PPARgamma[J].Gastroenterology,2007,132(5):1955-1967.
[9] Kim KH,Hong SP,Kim K,et al. HCV core protein induces hepatic lipid accumulation by activating SREBP1 and PPARgamma[J]. Biochem Biophys Res Commun,2007,355(4):883-888.
[10]河福金,贲长恩,王德福.小柴胡汤对大鼠酒精性肝损伤的防护作用[J].中西医结合肝病杂志,1994,4(3):19.
[11]尤圣富,郑培永,季光.清肝活血方对酒精性肝病大鼠肝脏脂质过氧化损伤的影响[J].中西医结合肝病杂志,2005,15(4):218-220.
[12]石伟珍,过建春,施军平,等.三七对酒精性肝病大鼠肝组织转化生长因子β表达的影响[J].中华中医药学刊, 2009,27(4):817-821.
[13]荀运浩,陈芝芸,施军平,等.酒精性肝病大鼠血清肿瘤坏死因子-α的动态变化及三七对其的影响[J].中华中医药学刊, 2009,27(5):1055-1058.
[14]García-Villafranca J,Guillén A,Castro J.Ethanol consumption impairs regulation of fatty acid metabolism by decreasing the activity of AMP-activated protein kinase in rat liver[J]. Biochimie,2008,90(3):460-466.
[15]You M, Matsumoto M, Pacold CM, et al. The role of AMP-activated protein kinase in the action of ethanol in the liver[J].Gastroenterology,2004,127(6):1798-808.
[16]Donohue TM Jr.Alcohol-induced steatosis in liver cells[J].World J Gastroenterol, 2007,13(37):4974-4978.