加味补阳还五汤对七氟烷麻醉处理后早期糖尿病肾病大鼠肾足细胞影响实验研究

Effect of the Modified Buyang Huanwu Decoction on the Early of the Diabetic Nephropathy Rats Podocyte after Anesthesia Induction with Sevoflurane Preconditioning

目的:探讨加味补阳还五汤(MDIY)对七氟烷麻醉处理后的早期糖尿病肾病(DN)大鼠肾足细胞的影响。方法:将早期DN大鼠随机分为4组(每组n=16):MDIY组(A组),罗格列酮组(B组),MDIY+罗格列酮组(C组)和模型对照组(D组),另设正常大鼠(E组)为空白对照组。连续治疗7周,每天收集大鼠死亡数和葡萄糖(GLU)、肌酐(CR)、尿素氮(BUN)和尿微量白蛋白(UAL)。七氟烷麻醉处理后采集标本,通过苏木精和伊红染色(H&E)收集肾脏和肾病理变化检查。应用实RT-PCR和western blotting检测评估肾足突细胞各m-RNA的表达水平,包括nephrin、podocin、α-SMA和desmin的表达。结果:在治疗周期中,A组、B组和C组的GLU和UAL水平均低于D组(P<0.01),且7周后3组的GLU、CR和BUN水平也都远低于D组(P<0.05)。其中,C组的上述生化指标远低于A组和B组(P<0.05)。相比于正常SD大鼠,通过H&E染色,观察各组的肾小球疾病、慢性肾小管间质损伤和肾小球足细胞损伤的发病率。然而,只有C组显示更能优化改善上述损伤的发病率。在A组、B组和C组与D组比较,α-SMA和desmin的表达减少,而nephrin和podocin的表达明显增加(P<0.05);C组不仅nephrin和podocin mRNA表达(P<0.05),而且在蛋白表达方面均观察有协同效应。结论:MDIY对早期DN引起的肾功能损害是一种保护作用,可以大大提高与罗格列酮相结合的治疗效果,其机制是通过增加nephrin、podocin的表达和减少α-SMA、desmin的表达,从而推测MDIY具有显著的协同治疗DN作用。七氟烷预处理能改善糖尿病肾病足细胞结构和功能,对围术期外科快速康复有利。

Objective:To expore the effects and mechanism of modified Buyang Huanwu Decoction reducing the risk of kidney damage caused by the early of the diabetic nephropathy(DN)with sevoflurane preconditioning.Methods:Then DN rats were randomly divided into four groups(n=16),MDIY(group A),rosiglitazone(group B),MDIY+rosiglitazone(group C)and normal saline(group D).During the treating cycle,the glucose(GLU)and urine albumin(UAL)were detected continuously.After the treatment of 7 weeks,the rats were sacrificed and the level of GLU,serum creatinine(SCR)and blood urea nitrogen(BUN)were detected in the serum.At the same time,the kidneys were collected and the renal pathological changes were inspected by hematoxylin and eosin(H&E)staining.The expression levels of proteinuria including nephrin,podocin,α-smooth muscle actin(α-SMA)and desmin were evaluated by Real-time PCR and Western blotting.Results:During the treating cycle,GLU and UAL in the A,B and C groups were lower than group D(P<0.01),after 7 weeks the level of GLU,CR and BUN levels in group A,B and C were much lower than group D(P<0.05).Compared with group A and group B,biochemical indexes above mentioned were much lower in group C(P<0.05).Incidences of glomerular disorders,chronic tubulointerstitial injure and glomerular podocyte damage were observed in group A,B,C and D when compared to normal SD rats by H&E staining.However,group C showed optimal improvement of glomerular disorders,chronic tubulointerstitial injure and glomerular podocyte damage.The expression ofα-SMA and desmin decreased,while the expression of nephrin and podocin increased markedly in group A,B and C comparing with group D(P<0.05),which were detected by Real-time PCR and Western blotting.The synergistic effect was observed not only in the mRNA expression of nephrin and podocin in group C(P<0.05),but also in the protein expression.Conclusion:MDIY was proven to be a protective effect in kidney damage caused by DN and could considerably ameliorate the therapeutic efficacy combined with rosiglitazone,which developed a treating role through increasing the expression of nephrin and podocin and decreasing the expression ofα-SMA and desmin.And sevoflurane preconditioning can improve the structure and function of diabetic nephropathy,and it is beneficial to the rapid rehabilitation of perioperative period.