棉酚对T2DM合并NAFLD大鼠肝脏的保护作用  

Heptoprotective effect of gossypol in type 2 diabetic rats complicated with non-alcoholic fatty liver disease

作　　者：王家员[1] 樊建霜[1] 吴亮[2] 黄卡特[2] 王蓉蓉[2] 

机构地区：[1]浙江中医药大学附属温州市中医院中药房,325015 [2]温州医科大学附属第一医院病理科

出　　处：《浙江医学》2017年第9期688-693,773,共7页Zhejiang Medical Journal

基　　金：温州市科技计划项目(Y20120010)

摘　　要：目的探讨棉酚对2型糖尿病(T2DM)合并非酒精性脂肪性肝病(NAFLD)大鼠肝脏的保护作用。方法将30只SD雄性大鼠随机分为棉酚组、T2DM组和正常组,每组10只。高脂高糖饮食喂养4周后腹腔注射小剂量(30mg/kg)链脲佐菌素构建T2DM大鼠模型。棉酚组用棉酚混悬液灌胃12周,每次剂量为15mg/kg,前4周为1次/d,后8周为1次/周。实验结束后检测大鼠血清ALT、AST、白蛋白(ALB)及透明质酸(HA)水平;透射电镜下观察肝组织超微结构改变;经HE染色、Masson胶原纤维染色后,在光镜下分别观察肝组织形态学改变、纤维化病变;免疫组化染色检测肝组织α-平滑肌肌动蛋白(α-SMA)、肝脏Ⅰ型11β类固醇脱氢酶(11β-HSD1)及糖皮质激素受体(GR)蛋白表达;采用半定量RT-PCR法检测肝组织葡萄糖-6-磷酸酶(G6Pase)、磷酸烯醇式丙酮酸羧激酶(PEPCK)、瘦素受体(Leptin R)、胰岛素受体(Insulin R)mRNA表达水平。结果与正常组相比,T2DM组血清ALT、AST、HA水平均明显升高(均P<0.05),ALB水平明显降低(P<0.01);肝脏α-SMA、11β-HSD1、GR蛋白表达均明显增强(均P<0.05);肝组织Leptin R、Insulin R mRNA表达均明显升高(均P<0.01);临床病理学表现主要为肝细胞水肿、脂肪变性、气球样变、炎症细胞浸润及纤维组织增生,肝组织内可见较多活化的肝星形细胞。与T2DM组相比,棉酚组血清ALT、AST、HA水平均明显降低(均P<0.05),ALB水平明显升高(P<0.05);肝脏11β-HSD1、GR蛋白表达均明显降低(均P<0.05);肝组织G6Pase、PEPCK、Insulin R、Leptin R mRNA表达均明显下降(均P<0.01);临床病理学表现主要为肝细胞水肿、脂肪变性、气球样变及炎症现象稍有减轻,肝内纤维组织明显减少,肝组织内活化的肝星形细胞数量明显减少。结论低剂量棉酚可明显减轻T2DM合并NAFLD大鼠纤维化病变,其机制可能与其抑制组织11β-HSD1、Leptin R、Insulin R mRNA表达有关。Objective To investigate the hepatoprotective effect of gossypol in type 2 diabetic(T2DM) rats complicated with non-alcoholic fatty liver disease(NAFLD). Methods Thirty male Sprague-Dauley rats were divided randomly into normal control group, T2DM group and gossypol-treated group with 10 in each group. After being fed with high-fat feeding for 4 weeks,the latter two groups were injected with strepozotocin(30mg/kg) intraperitoneally to induce T2 DM. The rats in gossypol treated group were given gossypol at a dose of 15mg·kg^(-1)·d^(-1)for 4 weeks by gavage, then at a dose of 15mg·kg^(-1)·d^(-1)until the end of week 12. At the end of the experiment all animals were sacrificed, the concentration of serum ALT, AST, ALB and HA were measured. The ultrastructural changes of liver were observed by transmission electron microscopy(TEM), the pathologic changes and fibrosis of liver were observed by light microscopy(LM) with HE staining and Masson staining respectively. Additionally, the activated hepatic stellate cells(HSCs) and the protein expression of 11 beta-hydroxysteroid dehydrogenase type 1(11β-HSD1)and glucocorticoid receptors(GR) was assayed by immunohistochemistry(IHC), the m RNA expression of phosphoenolpyruvate carboxykinase(PEPCK), glucose-6-phosphatase(G6Pase), insulin receptor and leptin receptor in liver tissue was assayed by semi-quantitative RT-PCR. Results The levels of serum ALT, AST, HA increased(P<0.05), serum ALB decreased(P<0.05), the11β-HSD1 and GR protein expression increased(P<0.05), the m RNA expression of insulin receptor, leptin receptor increased(P <0.05) in liver tissue of T2 DM group compared with normal control group. Clinical pathology showed obvious liver hydropicdegeneration, fatty degeneration, ballooning degeneration, inflammatory infiltration, fibrosis, a great number of activated HSCs in liver tissue of T2 DM group. The levels of serum ALT, AST, HA decreased(P <0.05), serum ALB increased(P <0.01), the11β-HSD1 and GR protein expression decreased(P <0.05), the mR NA expression

关 键 词：棉酚 2型糖尿病 非酒精性脂肪性肝病 Ⅰ型11β类固醇脱氢酶 瘦素受体 胰岛素受体 

分 类 号：R575[医药卫生—消化系统]