芪藤消浊颗粒对阿霉素肾病大鼠补体表达的影响

Protective effects of Qiteng Xiaozhuo Granules on complement expression in adriamycin-induced nephropathy rats

为探究芪藤消浊颗粒对慢性肾小球肾炎(chronic glomerulonephritis,CGN)大鼠补体表达的影响,予大鼠尾静脉注射阿霉素(adriamycin,ADR)制备CGN模型,用不同剂量的芪藤消浊颗粒(21.6 g/kg、10.8 g/kg、5.4 g/kg)连续灌胃治疗30 d,脲酶法检测血尿素氮(blood urea nitrogen,BUN)含量,肌氨酸氧化酶法检测血肌酐(serum creatinine,Scr)含量,ELISA检测胱抑素C(cystatin C,CysC)和视黄醇结合蛋白(retinol-binding protein,RBP)含量;H-E染色、六胺银染色、Masson染色、甲苯胺蓝染色和瑞氏-吉姆萨染色观察大鼠肾小球组织病变情况;Transwell细胞迁移实验观察大鼠肾组织对白细胞的招募能力;Western blotting及免疫组化法检测大鼠肾组织C1、C2、C3、C5和C5AR1蛋白表达水平;RT-qPCR检测CGN大鼠肾皮质组织中C1、C2、C3、C5和C5AR1 mRNA表达情况。结果显示,模型组大鼠肾小球萎缩严重且有明显的肾组织增生,周围有大量炎症细胞浸润;芪藤消浊颗粒可显著降低CGN大鼠血清中BUN、Scr、CysC和RBP含量,明显改善肾脏组织病理形态,降低肾脏对白细胞的招募能力,显著降低CGN大鼠肾组织中C1、C2、C3、C5、C5AR1 mRNA和C1、C2、C3、C5、C5AR1蛋白表达水平(均P<0.05)。因此,芪藤消浊颗粒治疗CGN的作用机制可能与调节补体表达水平有关。

To investigate the effects of Qiteng Xiaozhuo Granules on complement expression in rats with chronic glomerulonephritis(CGN),rats were injected with adriamycin(ADR)through tail vein to prepare CGN model.CGN rats were divided into three groups and given different doses of Qiteng Xiaozhuo Granules(21.6 g/kg,10.8 g/kg and 5.4 g/kg,respectively)by gavage for thirty consecutive days.The blood urea nitrogen(BUN)was determined by urease method;The serum creatinine(Scr)was determined by creatine oxidase method;The levels of cystatin C(CysC)and retinol-binding protein(RBP)were determined by ELISA.The renal pathology was monitored by H-E staining,PASM staining,Masson staining,Toluidine Blue staining,and Wright-Giemsa staining.The ability of renal tissue to recruit leukocytes was determined by Transwell cell migration assay.The protein expression levels of C1,C2,C3,C5,and C5 AR1 in rat kidney tissues were detected by Western blotting and immunohistochemistry and their mRNA levels in renal cortex were quantified by RT-qPCR.The results showed severe glomerular atrophy and obvious renal tissue hyperplasia,surrounded by a large number of inflammatory cells.Notably,the treatment of Qiteng Xiaozhuo Granules significantly reduced the levels of BUN,Scr,CysC and RBP in rat serum,greatly improved renal pathological morphology,and decreased the recruitment ability of kidney to leukocytes.The treatments also reduced the mRNA and protein levels of C1,C2,C3,C5 and C5AR1(all P<0.05)in rat kidney tissue.Above all,the protective effects of Qiteng Xiaozhuo Granules on CGN rats may due to its regulation of complement expression.