补阳还五汤对2型糖尿病大鼠肾组织Wnt蛋白表达影响研究

Effect of Buyang Huanwu Decoction on Wnt protein expression in renal tissue of type 2 diabetic rats

目的探讨补阳还五汤保护2型糖尿病大鼠肾脏的可能作用机制。方法取120只健康雄性SD大鼠,随机选择20只作为空白组,其余100只大鼠采用高脂饲料喂养和STZ注射法建立糖尿病模型。将造模成功大鼠随机分为模型组、阻断剂组、补阳还五汤组、补阳还五汤+阻断剂组,阻断剂组大鼠给予无翅型MMTV整合位点家族/β-连环蛋白(Wnt/β-catenin)信号通路阻断剂XAV-939腹腔注射,补阳还五汤组大鼠给予补阳还五汤灌胃,补阳还五汤+阻断剂组大鼠给予补阳还五汤灌胃并XAV-939腹腔注射,空白组和模型组给予等体积生理盐水灌胃,均连续干预4周。记录各组大鼠干预4周末体重,干预4周末腹主动脉取血,检测各组大鼠空腹血糖(FPG)、血肌酐(SCr)、血尿素氮(BUN)水平;处死各组大鼠后取肾脏,计算肾脏指数,苏木素-伊红染色(HE)观察肾脏病理变化,Western blot法检测肾脏组织中Wnt5a、Wnt4、β-catenin蛋白表达量。结果干预4周后,模型组大鼠体重明显低于空白组(P<0.05),FPG、BUN水平和肾脏指数均明显高于空白组(P均<0.05);肾组织病理观察显示大鼠肾组织结构紊乱,肾小球肥大,基质增生,系膜增厚,部分肾小管细胞出现空泡样变性;肾脏组织中Wnt5a、Wnt4、β-catenin蛋白相对表达量均明显高于空白组(P均<0.05)。与模型组比较,阻断剂组大鼠血清BUN水平、补阳还五汤+阻断剂组大鼠体重和补阳还五汤组大鼠FPG、BUN水平均明显降低(P均<0.05),补阳还五汤+阻断剂组大鼠血清SCr水平明显升高(P<0.05);阻断剂组、补阳还五汤组大鼠体重均明显高于补阳还五汤+阻断剂组(P均<0.05),而SCr水平明显低于补阳还五汤+阻断剂组(P均<0.05)。各给药组大鼠肾脏病理改变均有不同程度改善,补阳还五汤+阻断剂组改善最明显;阻断剂组大鼠肾脏组织中Wnt4蛋白相对表达量和补阳还五汤组、补阳还五汤+阻断剂组大鼠肾脏组织中Wnt5a、Wnt4、β-catenin蛋白相对表达量均明显降低(P均<0.05)。结论补阳还五汤可明显降低糖尿病大鼠血糖,减轻肾脏病变程度,机制可能与其抑制肾组织中Wnt蛋白的高表达有关。

Objective It is to explore the possible mechanism of Buyang Huanwu Decoction in protecting the kidney of type 2 diabetic rats.Methods One hundred and twenty healthy male SD rats were selected,in which 20 were randomly selected as blank group,and the remaining 100 rats were used to establish diabetes models by high-fat diet and STZ injection.The successful modeled rats were randomly divided into model group,blocker group,Buyang Huanwu Decoction group,Buyang Huanwu Decoction+blocker group,the rats in the blocker group were given the wingless MMTV integration site family/β-catenin(Wnt/β-catenin)signaling pathway blocker XAV-939 by intraperitoneal injection,the Buyang Huanwu Decoction group was given Buyang Huanwu Decoction by gavage,and the Buyang Huanwu Decoction+blocker group were given Buyang Huanwu Decoction by gavage and XAV-939 by intraperitoneal injection,and the blank group and the model group were given the same volume of normal saline by gavage,all groups were continuously treated for 4 weeks.The weight of rats in each group was recorded at the end of the 4 week intervention,and the blood of abdominal aorta was taken at the end of the 4 week intervention to detect the levels of fasting plasma glucose(FPG),serum creatinine(SCr)and blood urea nitrogen(BUN)in the rats of each group.After the rats in each group were sacrificed,their kidneys were taken out to calculate the renal index and observe the pathological changes by hematoxylin eosin staining(HE),the expression of Wnt5a,Wnt4 andβ-catenin protein in renal tissue was detected by Western blot method.Results After 4 weeks of intervention,the weight of rats in the model group was significantly lower than that in the blank group(P<0.05),and the levels of FPG and BUN were significantly higher than those in the blank group(all P<0.05);pathological observation of renal tissue showed that the structure of renal tissue in rats was disordered with glomerular hypertrophy,matrix proliferation,mesangial thickening,and some renal tubular cells showed vacuolar degeneration;the expression of Wnt5a,Wnt4 andβ-catenin protein was significantly higher than that in the blank group(P<0.05).Compared with the model group,the serum BUN level of the rats in the blocker group,the body weight of the rats in the Buyang Huanwu Decoction+blocker group,and the levels of FPG and BUN of the rats in the Buyang Huanwu Decoction group were significantly decreased(all P<0.05),the serum SCr level of the rats in the Buyang Huanwu Decoction+blocker group was significantly increased(P<0.05);the body weight of the rats in the blocker group and Buyang Huanwu Decoction group was significantly higher than that in Buyang Huanwu Decoction+blocker group,while the SCr level was significantly lower than that in Buyang Huanwu Decoction+blocker group(P<0.05).The pathological changes of the kidneys of the rats in each administration group were improved to different degrees,and the Buyang Huanwu Decoction+blocker group improved the most;the relative expressions of Wnt5a,Wnt4 andβ-catenin in kidney tissue of Buyang Huanwu decoction+blocker group were significantly decreased(all P<0.05).Conclusion Buyang Huanwu Decoction can significantly reduce blood glucose and reduce the degree of renal lesion in diabetes rats,and the mechanism may be related to its inhibition of the high expression of Wnt protein in renal tissue.