参芪蛭龙汤通过ERK/cPLA2信号通路对膜性肾病小鼠的治疗作用及机制研究  

Therapeutic effect and mechanism of Shenqi Zhilong Decoction on mice with membranous nephropathy through ERK/c PLA2 signaling pathway

作　　者：薛丕良 李丽琦 徐梅秀 李星宇 王顺 XUE Pi-liang;LI Li-qi;XU Mei-xiu;LI Xing-yu;WANG Shun(Heilongjiang Academy of Traditional Chinese Medicine,Harbin 150036;Heilongjiang University of Traditional Chinese Medicine,Harbin 150042,China)

机构地区：[1]黑龙江省中医药科学院,黑龙江哈尔滨150036 [2]黑龙江中医药大学,黑龙江哈尔滨150040

出　　处：《海南医学院学报》2022年第15期1127-1134,共8页Journal of Hainan Medical University

基　　金：黑龙江省自然科学基金项目(LH2020H104);黑龙江省博士后基金(LBH-Z20033)。

摘　　要：目的:观察参芪蛭龙汤方对膜性肾病(MN)小鼠的治疗作用及可能的机制。方法:通过多次尾静脉注射阳离子牛血清白蛋白制备MN模型小鼠。造模结束后,依据尿蛋白数值将MN小鼠随机分为模型对照组(等量蒸馏水)、参芪蛭龙汤低剂量组(12 g生药/kg)、参芪蛭龙汤高剂量组(24 g生药/kg)和雷公藤多苷片组(14 mg/kg)。另取10只正常小鼠作为空白对照组(等量蒸馏水)。连续给药4周,每天灌胃给药1次。给药结束后,收集24 h尿液,测定尿蛋白数值;眼球取血,测定肾功、肝功、血脂及血清IL-6、IL-4、TNF-α含量;HE染色观察肾脏病理变化;免疫组化染色观察肾脏IgG沉积情况;Western-blot法测定肾脏组织ERK1/2和cPLA2蛋白表达;RT-PCR法检测肾脏组织Neph-1、Nephrin及Podocin基因表达。结果:与模型组比较,参芪蛭龙汤组病理变化明显改善;参芪蛭龙汤低、高剂量组可显著降低MN小鼠尿蛋白数值;降低血脂TC及TG水平(P<0.05或P<0.01);升高ALB及TP水平(P<0.05或P<0.01);对CRE、UREA及UA水平无显著影响(P>0.05);降低血清IL-6、IL-4及TNF-α含量(P<0.05或P<0.01);显著下调MN小鼠肾脏组织中p-ERK1/2和p-cPLA2蛋白表达水平(P<0.05或P<0.01);显著升高肾脏组织Neph1、Nephrin及Podocin mRNA基因表达水平(P<0.01)。结论:参芪蛭龙汤对MN小鼠具有较好的治疗作用,作用机制与调节Nephrin-podocin-Neph1受体复合物相关基因表达,保护肾小球滤过屏障;抑制ERK/cPLA2途径的激活,减轻肾小球上皮细胞损伤,从而减少促炎细胞因子分泌有关。Objective:To explore the therapeutic effect and underlying mechanism of Shenqi Zhilong Decoction on mice with membranous nephropathy(MN).Methods:Mice with MN was established by injecting cationic bovine serum albumin(cBSA)into tail vein for several times.model mice were randomly divided into MN group(equal amount of distilled water),Shenqi Zhilong Decoction low dose group(12 g crude drug/kg),Shenqi Zhilong Decoction high dose group(24 g crude drug/kg),and Tripterygium wilfordii polyglycoside tablet group(14 mg/kg).Another 10 un-treatment mice were taken as control group(equal amount of distilled water).The drug was administered orally once a day for 4 weeks.After the last administration,24 h urine was collected to determine the urinary protein content;blood from inner canthus was collected to measure the changes of kidney function,liver function,blood lipid and levels of IL-6,IL-4 and TNF-αin serum in each group;HE staining was used to observe the pathological changes of kidney.Immunohistochemical staining was used to observe the expression of IgG in kidney.The protein expression of ERK1/2 and cPLA2 in renal tissues was determined by Western-blot method.The gene expression of Neph1,Nephrin and Podocin mRNA in kidney tissues were detected by RT-PCR.Results:Compared with model group,Shenqi Zhilong decoction at low-dose and high-dose could significantly reduce the value of urine protein in MN mice;Decreased TC and TG levels(P<0.05 or P<0.01);Increased the levels of ALB and TP in liver function(P<0.05 or P<0.01);has no significant effects on the levels of CRE,UREA and UA in renal function(P>0.05).Decreased the contents of IL-6,IL-4 and TNF-αin serum(P<0.05or P<0.01);Significantly down-regulated the protein expression levels of p-ERK1/2 and p-cPLA2 in kidney tissues of MN mice(P<0.05 or P<0.01);Significantly increased the expression levels of NephP1,Nephrin and Podocin mRNA in renal tissues(P<0.01).Conclusion:Shenqi Zhulong Decoction has a good therapeutic effect on MN mice,and the mechanism of action is related to

关 键 词：参芪蛭龙汤 膜性肾病 ERK/cPLA2信号通路 Nephrin-podocin-Neph1受体复合物 

分 类 号：R285.5[医药卫生—中药学]