赖氨大黄酸对胆汁淤积性肝纤维化模型大鼠的治疗作用及其分子机制

doi:10.13481/j.1671-587x.20150309

吉林大学学报(医学版) ›› 2015, Vol. 41 ›› Issue (03): 481-485.doi: 10.13481/j.1671-587x.20150309

赖氨大黄酸对胆汁淤积性肝纤维化模型大鼠的治疗作用及其分子机制

郝晓方¹, 张荣花¹, 王琳¹, 王梅梅¹, 甄永占², 章广玲^1,3, 陈静⁴

1. 华北理工大学基础医学院病原生物学与免疫学教研室, 河北唐山 063000;
2. 华北理工大学基础医学院组织学与胚胎学教研室, 河北唐山 063000;
3. 华北理工大学基础医学院唐山市慢性病临床基础研究重点实验室, 河北唐山 063000;
4. 华北理工大学生命科学院细胞生物学教研室, 河北唐山 063000

收稿日期:2014-09-11 发布日期:2015-08-01
通讯作者: 章广玲,教授,硕士研究生导师(Tel:0315-3725918,E-mail:zhguangling@aliyun.com);甄永占,副教授,硕士研究生导师(Tel:0315-3725918,E-mail:yongzhanzhen@126.com) E-mail:zhguangling@aliyun.com;yongzhanzhen@126.com
作者简介:郝晓方(1987-),女,河北省邯郸市人,在读理学硕士,主要从事肝纤维化发病机制和治疗方面的研究。
基金资助:
国家自然科学基金资助课题(81201281);河北省科技厅自然科学基金资助课题(C2012401037,H2012401030,H2013209180,H2013209040)

Therapeutic effect of rhein lysinate on liver fibrosis in cholestatic model rats and its molecular mechanisms

HAO Xiaofang¹, ZHANG Ronghua¹, WANG Lin¹, WANG Meimei¹, ZHEN Yongzhan², ZHANG Guangling^1,3, CHEN Jing⁴

1. Department of Pathogen Biology and Immunology, College of Basic Medical Sciences, North China University of Sciences and Technology, Tangshan 063000, China;
2. Department of Histology and Embryology, College of Basic Medical Sciences, North China University of Science and Technology, Tangshan 063000, China;
3. Tangshan Key Laboratory for Preclinical of Science and Technology and Basic Research on Chronic Diseases, School of Basic Medical Sciences, North China Universityof Science and Technology, Tangshan 063000, China;
4. Department of Cell Biology, College of Life Science, North China University of Science and Technology, Tangshan 063000, China

Received:2014-09-11 Published:2015-08-01

摘要/Abstract

摘要：

目的: 探讨赖氨大黄酸(RHL)对胆汁淤积性肝纤维化模型大鼠的治疗作用,阐明RHL治疗大鼠胆汁淤积性肝纤维化的分子机制。方法:将35只大鼠分为对照组、模型组、35和70 mg·kg^-1RHL治疗组及赖氨酸组,每组7只大鼠。通过胆总管结扎(BDL)方法建立胆汁淤积性大鼠肝纤维化模型。使用全自动生化分析仪检测血清中天门冬氨酸氨基转移酶(AST)和丙氨酸氨基转移酶(ALT)活性以及总胆汁酸(TBA)和总胆红素(TBIL)水平;HE染色对肝脏组织行病理学检查;Masson三色染色观察肝组织纤维数量;使用试剂盒通过酶标仪测定肝脏组织中羟脯氨酸(Hyp)水平;免疫组织化学染色检测肝脏组织中α平滑肌肌动蛋白(α-SMA)的分布;Western blotting法检测α-SMA表达水平。结果:与对照组比较,模型组大鼠肝脏质量与体质量比值、血清中AST和ALT活性以及TBA和TBIL水平升高(P<0.05);肝组织肝小叶结构被破坏,纤维异常增生,Hyp水平亦升高(P<0.05)。免疫组织化学染色,α-SMA着色于胞膜和胞浆且表达增多;Western blotting法,与对照组比较,模型组大鼠α-SMA表达水平升高(P<0.05)。与模型组比较,35和70 mg·kg^-1RHL治疗组大鼠肝脏质量与体质量比值、血清中AST和ALT活性以及TBA和TBIL水平明显降低(P<0.05)。肝脏病理组织学,与模型组比较,35和70 mg·kg^-1RHL治疗组大鼠肝脏组织中纤维成分数量减少,Hyp水平降低(P<0.05);免疫组织化学和Western blotting法,与模型组比较,35和70 mg·kg^-1 RHL治疗组大鼠肝脏组织中α-SMA表达减少(P<0.05)。结论:RHL能抑制肝星形细胞激活、肝脏纤维变性和蓄积以发挥对胆汁淤积性肝纤维化大鼠肝脏的保护作用,RHL有望成为治疗胆汁淤积性肝纤维化的药物。

关键词: 胆汁淤积, 肝纤维化, 赖氨大黄酸

Abstract:

Objective To investigate the therapeutic effect of rhein lysinate (RHL) on the liver fibrosis in the cholestatic model rats, and to expound the molecular mechanisms of RHL in treatment of liver fibrosis in the cholestatic rats. Methods 35 rats were divided into control group, model group, 35 and 70 mg·kg^-1 RHL treatment groups, and lysine group(n=7).The rat models of cholestatic hepatic fibrosis were established by bile duct ligation (BDL) method.The activities of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and the levels of total bile acid (TBA), total bilirubin (TBIL) were detected by using an automatic biochemical analyzer;the pathological changes of liver tissue of the rats in various groups were observed by HE staining;the content of fiber in liver tissue was observed by Masson's trichrome stain;the level of hydroxyproline (Hyp) was detected by microplate reader according to the instruction of respective kits;the expression of α-smooth muscleactin (α-SMA) was detected by immunohistochemical staining;the expression level of α-SMA was detected by Western blotting method. Results Compared with control group, the ratio of liver weight to body weight and the activities of AST and ALT, the levels of TBA and TBIL in serum of the rats in model group were increased (P<0.05);the hepatic lobule structure of hepatic tissue was destroyed, and the fibrous tissue hyperplasia was found, the Hyp level was also increased(P<0.05);the immunohistochemical staining Results showed that α-SMA expressed in cell membrane and cytoplasm, and the Western blotting Results showed that the expression level of α-SMA protein was increased(P<0.05). Compared with model group, the ratio of liver weight to body weight and the activities of AST, ALT and the levels of TBA, TBIL in serum in 35 and 70 mg·kg^-1 RHL groups were significantly decreased(P<0.05);the liver histopathological examination found that the fibers of the rats in 35 and 70 mg·kg^-1 RHL groups were reduced, and the levels of Hyp were also decreased(P<0.05); the immunohistochemistry and Western blotting Results showed that the levels of α-SMA protein were reduced(P<0.05). Conclusion The hepatic protection of RHL is correlated with the inhibition of the activation of hepatic stellate cells, fibrosis and fiber accumulation of liver tissue.It is concluded that RHL would be a therapeutic drug for the patients with cholestatic liver fibrosis.

Key words: cholestasis, liver fibrosis, rhein lysinate

中图分类号:

R512.6

郝晓方, 张荣花, 王琳, 王梅梅, 甄永占, 章广玲, 陈静. 赖氨大黄酸对胆汁淤积性肝纤维化模型大鼠的治疗作用及其分子机制[J]. 吉林大学学报(医学版), 2015, 41(03): 481-485.

HAO Xiaofang, ZHANG Ronghua, WANG Lin, WANG Meimei, ZHEN Yongzhan, ZHANG Guangling, CHEN Jing. Therapeutic effect of rhein lysinate on liver fibrosis in cholestatic model rats and its molecular mechanisms[J]. Journal of Jilin University Medicine Edition, 2015, 41(03): 481-485.

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赖氨大黄酸对胆汁淤积性肝纤维化模型大鼠的治疗作用及其分子机制

Therapeutic effect of rhein lysinate on liver fibrosis in cholestatic model rats and its molecular mechanisms

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