益肾胶囊对糖尿病肾病大鼠肾组织LC3Ⅱ、p62表达的影响

Effects of Yishen Capsule on the Expression of LC3Ⅱand p62 in Renal Tissue of Diabetic Nephropathy Rats

目的:探讨益肾胶囊对糖尿病肾病大鼠肾组织LC3Ⅱ、p62表达的影响。方法:将40只雄性SD大鼠,随机分为正常组、糖尿病肾病组、益肾胶囊组、白藜芦醇组,每组10只。益肾胶囊组给予益肾胶囊(0.625 g·kg^(-1)·d^(-1))灌胃、白藜芦醇组给予白藜芦醇(30 mg·kg^(-1)·d^(-1))灌胃、正常组和糖尿病肾病组每日给予等量生理盐水灌胃,时间为8周。观察大鼠一般状态,在0、4、8周末测定大鼠体重、血糖及24 h尿蛋白定量;并在8周末于大鼠腹主动脉取血,测定肌酐(Scr)、尿素(BUN)、胆固醇(TC)、三酰甘油(TG),用HE、PAS染色法分别观察肾脏组织病理变化,用免疫组化法测定自噬相关蛋白LC3Ⅱ及泛素结合蛋白p62(p62/SQSTMI)的表达变化。结果:(1)与正常组相比,糖尿病肾病组Scr、BUN、TC、TG、24 h尿蛋白定量均有所升高(P<0.05);与糖尿病肾病组相比,益肾胶囊组及白藜芦醇组Scr、BUN、TC、TG、24 h尿蛋白定量均有所下降(P<0.05);(2)与正常组相比,糖尿病肾病组镜下肾组织系膜基质增生较明显,肾小管管腔狭窄,上皮细胞水肿,呈现空泡样变性,足细胞自噬相关蛋白LC3Ⅱ表达减少(P<0.05),p62表达增多(P<0.05);与糖尿病肾病组相比,益肾胶囊组及白藜芦醇组肾组织病理改变明显减轻,足细胞自噬相关蛋白LC3Ⅱ的表达增加(P<0.05),p62表达减少(P<0.05)。结论:益肾胶囊可能通过改善糖尿病肾病大鼠肾组织的自噬障碍,从而对糖尿病肾病的肾脏发挥保护作用。

Objective:To explore the effect of Yishen capsule on the expression of LC3Ⅱand p62 in renal tissue of diabetic nephropathy rats.Methods:Forty male SD rats were randomly divided into four groups,namely the normal group,the diabetic nephropathy group,the Yishen capsule group and the resveratrol group,with 10 rats in each group.The Yishen capsule group were given Yishen capsule,at a dose of 0.625 g·kg^(-1)·d^(-1) for 8 weeks.And the resveratrol group were given resveratrol,at a dose of 30 mg·kg^(-1)·d^(-1) for 8 weeks.The normal group and the diabetic nephropathy group were given the same amount of normal saline for 8 weeks.Observe the general state changes of the rats,and measure the body weight,blood glucose and 24-hour urine protein of the four groups at the end of 0,4 and 8 weeks.At the end of 8 weeks,blood was taken from the abdominal aorta of the four groups to measure the related biochemical indexes such as cholesterol(TC),triglyceride(TG),serum creatinine(Scr)and blood urea nitrogen(BUN).The pathological changes of renal tissue were detected by HE and PAS staining.The expression of autophagy related protein LC3Ⅱand ubiquitin-binding protein p62(p62/SQSTMI)were detected by immunohistochemistry.Results:(1)Compared with the normal group,the quantitative values of serum creatinine,blood urea nitrogen,cholesterol,triglyceride and 24-hour urine protein in diabetic nephropathy group were increased(P<0.05).Compared with the diabetic nephropathy group,the quantitative values of serum creatinine,blood urea nitrogen,cholesterol,triglyceride and 24-hour urine protein in Yishen capsule group and resveratrol group were decreased(P<0.05).(2)Compared with the normal group,the mesangial matrix hyperplasia of diabetic nephropathy group was more obvious;the renal tubules narrowed,the epithelial cells of renal tubular were edematous,the renal tubules showed vacuolar degeneration,the expression of autophagy related protein LC3Ⅱwas decreased(P<0.05),the expression of autophagy related protein p62 was increased(P<0.05).Compared with the diabetic nephropathy group,the renal pathological changes of rats in Yishen capsule group and resveratrol group were significantly reduced,the expression of LC3Ⅱwas increased(P<0.05),and the expression of p62 was decreased(P<0.05).Conclusion:Yishen capsule may play a protective role in diabetic nephropathy by improving the autophagy of renal tissue in diabetic nephropathy rats.