贞清方对2型糖尿病大鼠非酒精性脂肪肝病变及肝脏盐诱导激酶1表达的影响

目的观察贞清方对2型糖尿病大鼠非酒精性脂肪肝的作用及其对大鼠肝脏盐诱导激酶1（salt-inducible kinase 1， SIK1）、固醇调节元件结合蛋白-1c（sterol-regulatory element binding protein-1c， SREBP-1c）表达的影响。方法应用高糖高脂饮食结合小剂量链脲佐菌素（streptozotocin， STZ）腹腔注射建立2型糖尿病大鼠模型，将成模大鼠随机分为模型组、贞清方组、二甲双胍组，每组8只，并设立正常对照组8只，贞清方组给予贞清方每日12 g生药/kg灌胃，二甲双胍组给予二甲双胍混悬液每日150 mg/kg灌胃，正常对照组及模型组以等体积蒸馏水灌胃。持续治疗12周。检测各组大鼠空腹血糖（FBG）、游离脂肪酸（FFA）、 血清甘油三酯（TG）、总胆固醇（TC）、 血清丙氨酸转移酶（ALT）及天冬氨酸转移酶（AST），称体重和肝脏湿重并计算肝重指数，测肝脏TG含量， HE染色和免疫组织化学法观察大鼠肝脏病理变化及SIK1在肝脏的表达，RT-PCR和Western blot检测各组大鼠肝脏组织SIK1、SREBP-1c mRNA和蛋白的表达。结果与正常对照组比较，模型组大鼠FBG、FFA、 TG、TC、ALT、AST水平、肝重指数和肝脏TG含量显著升高（P〈0.01），肝脏重度脂肪变性，肝SIK1 mRNA和蛋白表达明显减少（P〈0.01），SREBP-1c mRNA 和蛋白表达增多（P〈0.01）；药物治疗后，与模型组比较，贞清方组与二甲双胍组大鼠FBG、FFA、 TG、TC、ALT、AST及肝重指数均降低，肝脏TG含量下降，SIK1 mRNA 和蛋白表达明显增多，而SREBP-1c mRNA 和蛋白表达明显减少（P〈0.05，P〈0.01），同时大鼠的肝脏病变程度得到了改善。贞清方组FBG、TG、TC、ALT、AST、肝TC含量、SIK1 mRNA 和蛋白表达、SREBP-1c mRNA 和蛋白表达均优于二甲双胍组（均P〈0.05）。结论本实验发现2型糖尿病并发非酒精性脂肪肝大鼠肝脏SIK1表达降低，同时贞清方可减轻2型糖尿病大鼠非

Effect of Zhenqing Recipe on Non-alcoholic Fatty Liver in Type 2 Diabetes Rats and the Expression of SIK1

Objective To observe the effect of Zhenqing Recipe （ZQR） on non-alcoholic fatty liver （NAFL）, and the expression of hepatic salt-inducible kinase 1 （SIK1） and sterol-regulatory element binding protein-lc （SREBP-lc） in type 2 diabetes rats. Methods A rat model of type 2 diabetes was established by high fat/sucrose diet combined with intraperitoneal injection of small dose streptozotocin （STZ） . Modeled rats were randomly divided into the model group, the ZQR group, and the metformin group, 8 in each group. Eight rats were recruited as a normal control group. ZQR at the daily dose of 12 g crude drugs/kg was administered to rats in the ZQR group by gastrogavage. Metformin suspension at the daily dose of 150 mg/kg was administered to rats in the metformin group by gastrogavage. Equal volume of distilled water was administered to rats in the normal control group and the model group. All medication lasted for 12 weeks. The levels of fasting blood glucose （FBG）, free fatty acid （FFA）, serum triglyceride （TG）, serum total cholesterol （TC）, serum alanine aminotransferase （ALT）, and aspartate aminotransferase （AST） were detected. The body weight and wet liver weight were weighed, and the liver weight index calculated. The liver TG content was measured. The pathological changes of liver and the expression of SIK1 were observed by HE staining and immunohistochemistry. The mRNA and protein expression of SlK1 and SREBP-1c were detected using RT-PCR and Western blot. Results Compared with the normal control group, FBG, FFA, TG, TC, ALT, AST, liver weight index, and liver TG contents significantly increased （P 〈0.01 ） ; liver steatosis was severe, the mRNA and protein expression of SIK1 obviously decreased （P 〈0.01 ） ; mRNA and protein expression of SREBP-1c increased （P 〈0.01）. After drug therapy, compared with the model group, FBG, FFA, TG, TC, ALT, AST, and liver weight index significantly decreased, liver TG contents significantly decreased, the mRNA and protein e