火绒草黄芪混煎液对系膜增生性肾小球肾炎模型大鼠的改善作用及机制

Improvement effects of Huorongcaohuangqi admixture decoction on mesangial proliferative glomerulonephritis model rats and its mechanism

目的研究火绒草黄芪混煎液(HRCHQ)对系膜增生性肾小球肾炎(MsPGN)模型大鼠的改善作用及机制。方法将70只大鼠按体质量分为假手术组(n=10)和造模组(n=60)。假手术组大鼠行假手术,造模组大鼠采用免疫学方法(弗氏佐剂+牛血清蛋白+脂多糖)复制MsPGN模型。将造模成功的50只大鼠按体质量随机分为模型组和HRCHQ高、中、低剂量组(4.05、2.03、1.02g/kg)以及盐酸贝那普利片组(20 mg/kg),每组各10只。假手术组、模型组大鼠灌胃蒸馏水,其余各组大鼠灌胃相应药液15mL/kg,每天1次,连续5周。末次给药后,测定大鼠血清中总蛋白、白蛋白、尿素氮含量,采用免疫组织化学染色法检测大鼠肾脏组织中转移生长因子β1(TGF-β1)、肿瘤坏死因子α(TNF-α)的表达,采用免疫荧光法检测大鼠肾脏组织中补体C3的表达,采用Western blot法检测大鼠肾脏组织中磷脂酰肌醇3激酶/蛋白激酶B/糖原合酶激酶3β(PI3K/Akt/GSK3β)信号通路相关蛋白的磷酸化水平及纤连蛋白(FN)的表达水平。结果与模型组比较,HRCHQ高剂量组大鼠血清中总蛋白、白蛋白含量均显著升高(P<0.05或P<0.01);血清中尿素氮含量,肾脏组织中TGF-β1、TNF-α、FN、补体C3表达水平和PI3K、Akt、GSK3β蛋白的磷酸化水平均显著降低(P<0.05或P<0.01)。结论HRCHQ可通过减少系膜细胞增殖以及抑制PI3K/Akt/GSK3β信号通路活性来改善MsPGN。

OBJECTIVE To study the improvement effects of Huorongcaohuangqi(HRCHQ) admixture decoction on mesangial proliferative glomerulonephritis(MsPGN)model rats and its mechanism.METHODS Totally 70 rats were divided into sham operation group(n=10) and modeling group(n=60) according to body weight.Sham operation group underwent sham operation,and MsPGN model was induced by immunological method(Freund’s adjuvant+BSA+lipopolysaccharide) in modeling group.After successfully modeling,50 rats were randomly divided into model group,HRCHQ high-dose,medium-dose and lowdose groups(4.05,2.03,1.02 g/kg),Benazepril hydrochloride tablet group(20 mg/kg),with 10 rats in each group.Sham operation group and model group were given distilled water intragastrically;other groups were given relevant medicine intragastrically 15 m L/kg,once a day,for consecutive 5 weeks.After last administration,the contents of total protein,albumin and urea nitrogen were determined in the serum of rats.The expressions of transforming growth factor β1(TGF-β1)and tumor necrosis factor α(TNF-α) in renal tissue were detected by immunohistochemistry;the expression of complement C3 in renal tissue was detected by immunofluorescence.The phosphorylation level of phosphatidylinositol 3 kinase/protein kinase B/glycogen synthase kinase 3β(PI3K/Akt/GSK-3β) signal pathway related proteins and expression level of fibronectin(FN) in renal tissue were detected by Western blot.RESULTS Compared with model group,the serum contents of total protein and albumin were increased significantly in HRCHQ high-dose group(P<0.05 or P<0.01).The serum content of urea nitrogen,the expressions of TGF-β1,TNF-α,FN and complement C3,and the phosphorylation levels of PI3 K,Akt and GSK3β protein were all decreased significantly(P<0.05 or P<0.01).CONCLUSIONS HRCHQ can improve Ms PGN by reducing the proliferation of mesangial cells and inhibiting the activity of PI3 K/Akt/GSK3β signaling pathway.