肾叶山蚂蝗醇提对大鼠慢性肾功能衰竭的防治作用

Therapeutic Effect of Desmodium Renifolium on Chronic Renal Failure

目的为探究肾叶山蚂蝗醇提液干预对慢性肾功能衰竭(CRF)模型小鼠的改善作用。通过腺嘌呤诱导复制大鼠慢性肾功能衰竭模型,探讨肾叶山蚂蝗防治CRF的药效及作用机制。方法将SD大鼠随机分为正常组、模型组、右归丸组(3.6 g·kg^(-1))、醇提取物高剂量组(21.87 g生药·kg^(-1))、中剂量组(7.29 g生药·kg^(-1))、低剂量组(2.43 g生药·kg^(-1))。造模前5天进行预防性给药,第6天开始,模型组与各给药组大鼠上午给予1.0%腺嘌呤灌胃,下午进行药物治疗,持续15天。末次给药24 h后,采用折射法测定尿比重(SG);生化分析仪测定尿肌酐(Ucr)、24 h尿蛋白(PRO);细胞分析仪测定红细胞计数(RBC)、血小板数(Plt)、血红蛋白(HGB)、平均血小板体积(Mpv)、平均红细胞容积(Mcv)。酶联免疫吸附试剂盒测定尿素氮(BUN)、血清肌酐(Scr)含量与尿α1-微球蛋白(α1-MG)、肾脏损伤分子-1(KIM-1)表达。苏木精-伊红染色观察大鼠肾脏病理形态、免疫组化染色和Western blot分析水通道蛋白2(AQP2)、转化生长因子-β1(TGF-β1)、低氧诱导因子(HIF-1α)相关蛋白表达。结果与模型组相比,给药后对大鼠精神萎靡、反应迟缓,毛色发黄粗糙伴脱落等各项症状和肾脏组织病理形态都有明显改善。血常规指标RBC、HGB、Mpv、Mcv水平升高,Plt水平降低。24 h总尿量显著下降(P<0.05),低剂量组SG明显上升(P<0.05)。当URO=3.2μmol·L-1时,药物治疗后尿PRO可恢复阴性。肾功能损伤指标BUN、α1-MG、KIM-1水平明显降低(P<0.05,P<0.01,P<0.001);给药组大鼠肾指数和Ccr水平明显改善(P<0.05,P<0.01)。同时相关AQP2、TGF-β1和HIF-1α蛋白表达均有所改善(P<0.01,P<0.001)。结论肾叶山蚂蝗醇提液可以改善慢性肾功能衰竭大鼠肾脏组织病理及各项指标,减轻肾损伤及肾纤维化程度,对于防治大鼠CRF具有积极作用。

Objective In order to explore the preventive effect and potential mechanism of Dai medicine Shenyeshan leech on chronic renal failure(CRF).The rat model of chronic renal failure was induced and replicated by adenine,and the pharmacodynamics and mechanism of CRF prevention and treatment were discussed.Methods SD rats were randomly divided into normal group,model group,Yougui pill group,high-dose alcohol extract group,middle-dose group,low-dose group.Prophylactic administration was performed 5 days before modeling,and starting from the 6th day,rats in the model group and each administration group were given 1.0%adenine by gavage in the morning,and drug treatment was given in the afternoon for 15 days.24 hours after the last administration,urine specific gravity(SG)was determined by refraction method;urine creatinine(Ucr)and 24-hour proteinuria(PRO)were determined by biochemical analyzer;red blood cell count(RBC),platelet number(Plt)were determined by cell analyzer,hemoglobin(HGB),mean platelet volume(Mpv),mean corpuscular volume(Mcv).Enzyme-linked immunosorbent assay kits were used to determine blood urea nitrogen(BUN),serum creatinine(Scr)content and urinaryα1-microglobulin(α1-MG),kidney injury molecule-1(KIM-1)expression.Hematoxylin-eosin staining was used to observe the pathological morphology of rat kidneys,and immunohistochemical staining was used to analyze the expression of aquaporin 2(AQP2),transforming growth factor-β1(TGF-β1),and hypoxia-inducible factor(HIF-1α)-related proteins.Results Compared with the model group,the rats were significantly improved in various symptoms such as lethargy,sluggish reaction,yellow and rough coat with falling off,and the pathological morphology of the kidneys after administration.Blood routine indexes RBC,HGB,Mpv,Mcv levels increased,Plt level decreased.The 24-hour total urine volume decreased significantly(P<0.05),and the SG in the low-dose group increased significantly(P<0.05).When URO=3.2μmol·L-1,urinary PRO could be recovered to negative after drug treatment.The levels of renal function injury indexes BUN,α1-MG and KIM-1 were significantly decreased(P<0.05,P<0.01,P<0.001);the renal index and Ccr levels of the rats in the administration group were significantly improved(P<0.05,P<0.01).At the same time,the related AQP2,TGF-β1 and HIF-1αprotein expressions were improved.To sum up,it can be seen that the leech of the kidney leaf mountain can improve the renal histopathology and various indicators of chronic renal failure rats,reduce the degree of renal injury and renal fibrosis,and has a positive effect on the prevention and treatment of CRF in rats.