龙胆苦苷对大鼠腹膜纤维化的抑制作用及其机制研究  

Inhibitory effect of gentiopicroside on peritoneal fibrosis in rats and its mechanism

作　　者：李玉博 张健[3] 张再媛 李城明 龙盘 肖雯婧 呼永河 LI Yu-bo;ZHANG Jian;ZHANG Zai-yuan;LI Cheng-ming;LONG Pan;XIAO Wen-jing;HU Yong-he(College of Integrated Traditional Chinese and Western Medicine,Southwest Medical University,Luzhou 646000,China;Laboratory of Basic Medicine,The General Hospital of Western Theater Command,Chengdu 610083,China;College of Medicine,Southwest Jiaotong University,Chengdu 610031,China;Chengdu University of Traditional Chinese Medicine,Chengdu 610075,China;Department of Ophthalmology,The General Hospital of Western Theater Command,Chengdu 610083,China;Department of Pharmacy,The General Hospital of Western Theater Command,Chengdu 610083,China)

机构地区：[1]西南医科大学中西医结合学院,四川泸州646000 [2]中国人民解放军西部战区总医院基础医学实验室,四川成都610083 [3]西南交通大学医学院,四川成都610031 [4]成都中医药大学,四川成都610075 [5]中国人民解放军西部战区总医院眼科,四川成都610083 [6]中国人民解放军西部战区总医院药剂科,四川成都610083

出　　处：《中国病理生理杂志》2022年第6期1075-1082,共8页Chinese Journal of Pathophysiology

基　　金：四川省科技厅项目(No.2019YFS0542)。

摘　　要：目的:观察龙胆苦苷对腹膜透析大鼠腹膜纤维化组织氧化应激反应和转化生长因子β(TGF-β)/Smads信号通路的影响,探讨龙胆苦苷防治腹膜纤维化的作用机制。方法:通过4.25%高糖腹膜透析液(100 mL/kg;每天1次)联合脂多糖(0.6 mg/kg;第1、3、5、7天注射)构建大鼠腹膜纤维化模型。将30只雄性SD大鼠随机分为5组:生理盐水组、模型组及低、中、高剂量(30、60和120 mg/kg)龙胆苦苷组,每组6只。透析28 d后行1 h腹膜功能试验:测定超滤量、初始腹透液与透出液葡萄糖比值、透析液与血浆尿素氮比值及葡萄糖转运量。取大鼠壁层腹膜进行腹膜病理形态学观察,计算腹膜厚度。取腹主动脉血检测血清丙二醛(MDA)、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)水平。免疫组化检测各组大鼠腹膜组织中α-平滑肌肌动蛋白(α-SMA)、上皮钙黏素(E-cadherin)和I型胶原(COL I)的表达情况。Western blot检测各组大鼠腹膜组织中TGF-β1、Smad3和Smad7蛋白的表达。结果:透析28 d后,经腹腔注射4.25%高糖腹膜透析液及脂多糖建立的腹膜纤维化大鼠模型腹膜功能显著降低(P<0.05),腹膜厚度显著增加(P<0.01),并可见炎症细胞浸润、纤维细胞及血管增生明显增多;大鼠血清中MDA含量显著增多,SOD和GSH-Px活性显著降低(P<0.01);免疫组化显示,α-SMA和COL I蛋白表达显著增加,E-cad⁃herin蛋白表达显著降低(P<0.01);Western blot显示,TGF-β1和Smad3蛋白表达显著增加,Smad7蛋白表达显著降低(P<0.01)。龙胆苦苷干预能够显著降低氧化应激水平,抑制TGF-β1和Smad3蛋白的表达,提高Smad7蛋白表达水平,且呈剂量依赖性(P<0.05)。结论:龙胆苦苷可减轻高糖腹透液联合脂多糖诱导的腹膜纤维化模型大鼠的腹膜纤维化病变程度,其机制与抑制氧化应激反应及腹膜间皮-间充质转化进程有关。AIM:To observe the effect of gentiopicroside on oxidative stress and transforming growth factor-β(TGF-β)/Smads signaling pathway in peritoneal fibrosis tissue of rats undergoing peritoneal dialysis,and to explore the mechanism of gentiopicroside in preventing and treating peritoneal fibrosis.METHODS:The rat model of peritoneal fi⁃brosis was established by 4.25%high glucose peritoneal dialysate(100 mL/kg,once a day)combined with lipopolysac⁃charide(0.6 mg/kg)injection on the 1st,3rd,5th and 7th days.Male SD rats were randomly divided into 5 groups:saline group,model group,and low-,middle-and high-dose(30,60 and 120 mg/kg)gentiopicroside groups,with 6 rats in each group.Peritoneal function test was performed after 28 d of dialysis,and ultrafiltration volume,initial dialysate/final dialysate glucose ratio,dialysate/plasma urea nitrogen ratio and glucose transport volume were measured.The parietal peritoneum was taken for peritoneal pathomorphological observation,and peritoneal thickness was calculated.The blood of abdominal aorta was taken for detecting serum levels of malondialdehyde(MDA),superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px).The expression ofα-smooth muscle actin(α-SMA),E-cadherin and collagen type I(COL I)in peritoneal tissues was detected by immunohistochemistry.Western blot was used to detect the expression of TGF-β1,Smad3 and Smad7 in peritoneal tissues.RESULTS:After 28 d of dialysis,the peritoneal function of the rat model of peritoneal fibrosis was significantly decreased(P<0.05),while the peritoneal thickness,inflammatory cell infiltration,fibrous cells and vascular proliferation were significantly increased.The serum MDA increased significantly,while SOD and GSH-Px decreased significantly(P<0.01).Immunohistochemistry showed that the protein expression ofα-SMA and COL I increased significantly,while the protein expression of E-cadherin decreased significantly(P<0.01).Western blot showed that the expression of TGF-β1 protein and Smad3 protein increased significantly,while the

关 键 词：龙胆苦苷 腹膜纤维化 氧化应激 间皮-间充质转化 

分 类 号：R285.5[医药卫生—中药学] R363.2[医药卫生—中医学]