参芪蛭龙汤联合雷公藤多苷片对膜性肾病模型大鼠保护作用的实验研究
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篇名: 参芪蛭龙汤联合雷公藤多苷片对膜性肾病模型大鼠保护作用的实验研究
TITLE: Experimental Study on the Protective Effect of Shenqi Zhilong Decoction Combined with Tripterygium Polygly- coside Tablets on Membranous Nephropathy Model Rats
摘要: 目的:探讨参芪蛭龙汤联合雷公藤多苷片对膜性肾病(MN)模型大鼠的保护作用及可能机制。方法:采用皮下注射结合尾静脉注射阳离子牛血清白蛋白+不完全费氏佐剂乳化液的方法复制MN大鼠模型,造模周期为6周。于造模第3周,根据24h尿蛋白定量(UTP)和体质量将造模大鼠随机分为模型对照组,参芪蛭龙汤低、高剂量组(4、8g/kg,按生药总量计),雷公藤多苷片组(9mg/kg)以及参芪蛭龙汤低、高剂量+雷公藤多苷片组(剂量与各单药组相同),每组10只;另取10只未造模的大鼠作为空白对照组。空白对照组大鼠灌胃等体积水,各给药组大鼠灌胃相应药物,每日1次,连续4周。于末次给药前1天,检测各组大鼠24hUTP;于末次给药后1h,检测各组大鼠血常规指标[白细胞(WBC)、红细胞(RBC)、血小板(PLT)计数]、肝功能指标[总蛋白(TP)、白蛋白(ALB)、天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)]、血脂指标[胆固醇(TC)、三酰甘油(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)]以及葡萄糖(GLU)、尿素氮(BUN)、血肌酐(Scr)含量;采用醋酸铀-柠檬酸铅染色法观察大鼠肾脏超微结构;采用免疫组织化学法检测大鼠肾组织中转化生长因子β(1TGF-β1)、乙酰肝素酶1(HPA-1)蛋白的表达情况。结果:与空白对照组比较,模型对照组大鼠肾组织中肾小球足突广泛融合或消失,可见微绒毛形成、基底膜重度增厚,上皮下可见大量电子致密物沉积,TGF-β1、HPA-1阳性细胞明显增多;其24hUTP,PLT计数,TC、TG、HDL-C含量以及TGF-β1、HPA-1阳性细胞百分比均显著升高,RBC计数和TP、ALB含量均显著降低(P<0.05或P<0.01)。与模型对照组比较,各给药组上述超微结构变化均有不同程度的改善,TGF-β1、HPA-1阳性细胞均有所减少;其24hUTP和TGF-β1阳性细胞百分比(参芪蛭龙汤低剂量组除外)、WBC计数(参芪蛭龙汤单用及联用组除外)、PLT计数和TC含量(雷公藤多苷片组除外)、TG含量(参芪蛭龙汤低剂量单用以及与雷公藤多苷片联用组除外)、HPA-1阳性细胞百分比均显示降低,且参芪蛭龙汤高剂量+雷公藤多苷片组TGF-β1阳性细胞百分比以及参芪蛭龙汤低、高剂量+雷公藤多苷片组HPA-1阳性细胞百分比均显著低于雷公藤多苷片组;RBC计数和GLU含量(参芪蛭龙汤低剂量组和雷公藤多苷片组除外)、TP和ALB含量(参芪蛭龙汤低剂量组除外)均显著升高,且参芪蛭龙汤高剂量+雷公藤多苷片组GLU含量显著高于雷公藤多苷片组(P<0.05或P<0.01)。结论:参芪蛭龙汤与雷公藤多苷片联用可缓解雷公藤多苷片的骨髓抑制作用,减少MN模型大鼠蛋白尿,并改善其肾脏组织的病理损伤;上述作用可能与其下调肾脏组织中TGF-β1、HPA-1蛋白表达,降低血液中TC、TG含量有关。
ABSTRACT: OBJECTIVE:To investigate the protective effect an d possible m echanism of Shenqi zhilong decoction (SZD) combined with Tripterygium polyglycoside tablets (TPT)on membranous nephropathy (MN)model rats. METHODS :MN rat model was established by subcutaneous and caudal vein injecting cationic bovine serum albumin + incomplete Freund adjuvant emulsion for 6 weeks. At the 3rd week of modeling ,model rats were randomly divided into model control group ,SZD low-dose and high-dose groups (4,8 g/kg,by total crude drugs ),TPT group (9 mg/kg),low-dose and high-dose of SZD+TPT groups (same dose as single group ),with 10 rats in each group according to 24 h UTP and weight. Another 10 rats withoutmodeling were taken as blank control group. Blank control group was given equal amount of water intragastrically administration groups were given relevant medicine intragastrically,once a day ,for consecutive 4 weeks. The 24 h UTP of rats were detected one day before the last administration;1 h after the la st administration ,blood routine 中indexes(WBC,RBC,PLT),liver func tion indexes (TP,ALB,AST,ALT),blood lipid indexes (TC,TG,HDL-C,LDL-C), the contents of glucose (GLU),urea nitrogen (BUN)and serum creatinine (Scr)were detected in each group. Uranyl acetate-lead citrate staining was used to observe ultrastructural changes of renal tissue. Immunohistochemical method was used to detect the protein expression of TGF-β1 and HPA- 1 in renal tissue. RESULTS :Compared with blank control group ,in the model control group,the glomerular podocytes were widely fused or disappeared ,microvilli were formed ,basement membrane was heavily thickened,a large number of electron dense substance was deposited under the epithelium ,TGF-β1 and HPA- 1 positive cells were significantly increased ;24 h UTP ,PLT,the contents of TC ,TG and HDL-C ,the percentage of TGF-β1 and HPA- 1 positive cells were increased significantly ,while RBC ,the contents of TP and ALB were decreased significantly (P<0.05 or P<0.01). Compared with model control group ,above ultrastructural changes of administration groups were improved to different extents ,and TGF-β1 and HPA- 1 positive cells were decreased. The 24 h UTP ,the percentage of TGF-β1 positive cells (except for SZD low-dose group),WBC(except for SZD alone groups and combination groups ),PLT and TC content (except for TPT group ),TG content (except for SZD low-dose alone and its combination group ),the percentage of HPA- 1 positive cells were decreased significantly ; the percentage of TGF-β1 positive cells in SZD high-dose+TPT group as well as the percentage of HPA- 1 positive cells in SZD+TPT groups were significantly lower than TPT group. RBC and GLU content (except for SZD low-dose group and TPT group ),TP and ALB content (except for SZD low-dose group )were increased significantly ,while the content of GLU in SZD high-dose+TPT group was significantly higher than TPT tablets group (P<0.05 or P<0.01). CONCLUSIONS :SZD combined with TPT can relieve myelosuppression caused by TPT ,reduce proteinuria of MN model rats and improve pathological damage of renal tissue in rats. Its mechanism is related to the down-regulation of protein expression of TGF-β1 and HPA- 1,and the reduction of TC and TG content in the blood.
期刊: 2020年第31卷第13期
作者: 薛丕良,李丽琦,刘欣欣,白茹,肖洪彬,牛雯颖
AUTHORS: XUE Piliang ,LI Liqi,LIU Xinxin ,BAI Ru,XIAO Hongbin ,NIU Wenying
关键字: 膜性肾病;参芪蛭龙汤;雷公藤多苷片;血常规指标;肝功能;血脂;转化生长因子β1;乙酰肝素酶1;大鼠
KEYWORDS: Membranous nephropathy ;Shenqi zhilong decoction ;Tripterygium polyglycoside tablets ;Blood routine indexes ;
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