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目的:观察益肾汤对糖尿病肾病(DN)大鼠内皮素-1(ET-1)和转化生长因子-β1(TGF-β1)的影响,并观察肾脏病理形态学变化,以探讨其改善糖尿病肾病的作用机制。方法:将50只SD雄性大鼠,随机分为5组,对照组、模型组、益肾汤大剂量组、益肾汤小剂量组、洛汀新组,每组10只。对照组行左肾摘除假手术,其余4组摘除左肾。1周后,各组大鼠每天灌胃给药,益肾汤大剂量组灌胃1.0 g/mL;益肾汤小剂量组灌胃0.5 g/mL;洛汀新组0.33 mg/mL。对照组和模型组灌等体积的生理盐水。分别在第4、8周检测24h尿微量白蛋白,心脏采血检测血肌酐(SCr)和尿素氮(BUN)。HE染色肾组织送病理科评价病变程度;免疫组化法测定ET-1及TGF-β1蛋白的表达情况。结果:1各组大鼠BUN、SCr及24 h尿微量白蛋白的比较:4周末时,模型组各指标与对照组比较,差异有非常显著性意义(P<0.01)。各给药组与模型组比较,差异均有显著性意义(P<0.05),洛汀新组与益肾汤大、小剂量组比较,差异均无显著性意义(P>0.05);第8周末时,模型组与对照组比较,差异有非常显著性意义(P<0.01),各给药组与模型组比较,差异有显著性或非常显著性意义(P<0.05,P<0.01);洛汀新组与益肾汤大、小剂量组比较,差异均有显著性或非常显著性意义(P<0.05,P<0.01),益肾汤大、小剂量组比较,差异均有显著性意义(P<0.05)。2肾组织病理改变:肾小球增生肥大、基底膜增厚、系膜基质增生减轻,肾小囊腔轻度狭窄,治疗8周后益肾汤组较同期模型组减轻,肾间质炎症细胞浸润减少。3各组大鼠肾组织TGF-β1和ET-1表达比较:4周时模型组TGF-β1、ET-1表达水平较对照组升高(P<0.05),各给药组与模型组比较,差异均有显著性意义(P<0.05)。8周时模型组大鼠肾组织中表达与对照组比较,差异均有非常显著性意义(P<0.01),益肾汤大剂量组与模型组比较,差异均有非常显著性意义(P<0.01),益肾汤大剂量组表达减少;益肾汤小剂量组、洛汀新组表达水平均高于益肾汤大剂量组(P<0.01),益肾汤小剂量组与洛汀新组无差异(P>0.05)。结论:益肾汤可通过减少TGF-β1和ET-1在DN模型组肾组织中的表达,减轻DN大鼠的肾脏组织损伤,提示益肾汤发挥肾脏保护作用的机制可能部分的是通过调节TGF-β1和ET-1的表达实现的。
Objective: To observe the effect of yishen decoction on the expression of ET-1 and TGF-β1 in rats with diabetic nephropathy (DN) The mechanism of nephropathy. Methods: Fifty SD male rats were randomly divided into five groups: control group, model group, Yishen Decoction high dose group, Yishen Decoction low dose group and Lotensin group, with 10 rats in each group. Control group left kidney removal of sham operation, the remaining 4 groups were removed left kidney. One week later, the rats in each group were given gavage daily. The high-dose Yishen decoction group was given gavage 1.0 g / mL, the Yishen decoction low dose group was gavage 0.5 g / mL, and the Lotensin group 0.33 mg / mL. The control group and model group were infused with equal volume of saline. Urinary microalbuminuria was detected at 4 and 8 weeks, and serum creatinine (SCr) and blood urea nitrogen (BUN) were detected by blood sampling in the heart. The pathological changes of HE staining and pathology were evaluated. The expression of ET-1 and TGF-β1 protein was detected by immunohistochemistry. Results: 1 Comparison of BUN, SCr and 24-hour urine microalbuminuria in each group: At the end of 4weeks, there were significant differences between the model group and the control group (P <0.01). There was no significant difference between the two groups (P> 0.05), and there was no significant difference between the two groups (P> 0.05) There was significant difference between the model group and the control group at the weekend (P <0.01). The difference between the model group and the model group was significant or very significant (P <0.05, P <0.01). There were significant differences between the two groups (P <0.05, P <0.01), and there was significant difference between YST group and Yishen decoction group (P <0.05) Significance (P <0.05). Renal pathological changes: glomerular hyperplasia hypertrophy, thickening of the basement membrane, mesangial matrix hyperplasia, mild renal cystic cavity stenosis, after 8 weeks of treatment Yishen decoction group than the same period model group decreased interstitial inflammatory cell infiltration . The expression of TGF-β1 and ET-1 in renal tissue of rats in each group was significantly higher than that in control group at 4 weeks (P <0.05). Compared with model group, the expression of TGF-β1 and ET- , The differences were significant (P <0.05). There was significant difference between the model group and the control group at 8 weeks (P <0.01), and there was significant difference between the Yishen decoction group and the model group (P < 0.01). The expression of Yishen Decoction in high dose group was lower than that in Yishen Decoction (P <0.01), Yishen Decoction low dose group and Lotensin No difference (P> 0.05). Conclusion: Yishen Decoction can reduce the damage of kidney in DN rats by decreasing the expression of TGF-β1 and ET-1 in the kidney of DN model rats, suggesting that the mechanism of Yishen Decoction can play a protective role in kidney partly by regulating TGF-β1 and ET-1 expression achieved.