加味参苓白术散对不同含糖量腹膜透析液诱导大鼠腹膜纤维化的干预作用  

Intervention of modified Shenling Baizhu San on peritoneal fibrosis induced by peritoneal dialysate with different sugar concentration in rats

作　　者：吕勇[1] 陈雪莉 张磊[1] 尚丽雯 王亿平[1] Lyu Yong;Chen Xueli;Zhang Lei;Shang Liwen;Wang Yiping(Department of Nephrology,the First Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230038,Anhui,China;Graduate School,Anhui University of Chinese Medicine,Hefei 230031,Anhui,China)

机构地区：[1]安徽中医药大学第一附属医院肾内科,安徽合肥230038 [2]安徽中医药大学研究生院,安徽合肥230031

出　　处：《中华危重病急救医学》2023年第8期875-880,共6页Chinese Critical Care Medicine

基　　金：国家自然科学基金(81673931);安徽高校自然科学研究重点项目(KJ2021A0566);安徽中医药大学临床科学重点研究项目(2020yfyzc13);全国中医临床优秀人才研修项目(2017-124)。

摘　　要：目的:探讨加味参苓白术散对不同含糖量腹膜透析液(PDF)诱导腹膜纤维化大鼠腹膜组织病理学改变和细胞外基质(ECM)的影响及其机制。方法:将70只雄性SD大鼠按照单纯随机抽样原则分为对照组、不同含糖量PDF模型组及其相应中药干预组,每组10只。分别腹腔注射含糖量为1.5%、2.5%、4.25%的PDF 100 mL·kg-1·d-1诱导大鼠腹膜纤维化模型,每日1次,连续8周;对照组大鼠给予等量生理盐水。各中药干预组大鼠每日于制模后即刻灌胃加味参苓白术散溶液10 mL/kg(每升含2.014 kg生药);各PDF模型组及对照组大鼠灌胃等量生理盐水。8周后,检测各组大鼠腹膜超滤量。取腹膜组织,苏木素-伊红(HE)染色后,光镜下观察壁层腹膜形态学改变及腹膜厚度;Masson染色后,光镜下观察腹膜胶原纤维沉积情况;采用蛋白质免疫印迹试验(Western blotting)测定腹膜组织中ECM主要成分E-钙黏蛋白(E-cadherin)、α-平滑肌肌动蛋白(α-SMA)和波形蛋白(Vimentin)的表达;采用免疫组化法检测ECM调控因子基质金属蛋白酶-9(MMP-9)、金属蛋白酶组织抑制剂-1(TIMP-1)和转化生长因子-β1(TGF-β1)的阳性表达。结果:与对照组比较,不同含糖量PDF均可诱导大鼠发生腹膜纤维化,且纤维化程度随含糖量增加而逐渐加重,表现为腹膜增厚,胶原纤维沉积增加,腹膜超滤量减少,伴随腹膜组织中E-cadherin、MMP-9表达下调,α-SMA、Vimentin、TIMP-1、TGF-β1表达上调,以4.25%PDF模型组腹膜组织病理学改变及ECM聚积情况更加严重。经加味参苓白术散干预后,与相应PDF模型组比较,大鼠腹膜纤维化均得到不同程度改善,仍以4.25%PDF相应中药干预组效果更为显著,壁层腹膜明显变薄(μm:101.86±16.01比140.65±10.13,P<0.05),胶原纤维沉积明显减少,腹膜超滤量明显增加(mL:-0.01±3.45比-3.53±1.84,P<0.05),同时腹膜组织中E-cadherin、MMP-9表达明显上调〔E-cadherin蛋白(E-cadherin/β-actin):0.84±0.08�Objective To investigate the effect of modified Shenling Baizhu San on the pathological changes and extracellular matrix(ECM)in rats with peritoneal fibrosis induced by peritoneal dialysate fluid(PDF)with different sugar concentration and its mechanism.Methods Seventy male Sprague-Dawley(SD)rats were randomly divided into control group,different sugar content PDF model groups and corresponding traditional Chinese medicine intervention groups,with 10 rats in each group.Peritoneal fibrosis model was reproduced by intraperitoneal injection of 100 mL·kg-1·d-1 PDF containing 1.5%,2.5%and 4.25%sugar once a day for 8 weeks.The rats in the control group were given the same amount of normal saline.The rats in the traditional Chinese medicine intervention groups were treated with gavage of 10 mL/kg of modified Shenling Baizhu San(containing 2.014 g crude drug per liter)immediately after modeling.The PDF model groups and the control group were given the same amount of normal saline by gavage.After 8 weeks,the peritoneal ultrafiltration volume of rats in each group was measured.The peritoneal tissues were collected and stained with hematoxylin-eosin(HE),and the structural changes and thickness of the parietal peritoneum were observed under a light microscope.After Masson staining,the deposition of collagen fibers was observed under a light microscope.Western blotting was used to detect the protein expressions of E-cadherin,α-smooth muscle actin(α-SMA)and Vimentin,the main components of ECM in parietal peritoneum.The positive expressions of matrix metalloproteinase-9(MMP-9),tissue inhibitor of metalloproteinase-1(TIMP-1),and transforming growth factor-β1(TGF-β1)were detected by immunohistochemical staining.Results Compared with the control group,PDF with different sugar contents could induce peritoneal fibrosis in rats,and the degree of fibrosis was gradually aggravated with the increase of sugar content,which was manifested as peritoneal thickening,increased collagen fiber deposition,decreased peritoneal ultrafiltratio

关 键 词：加味参苓白术散 腹膜纤维化 腹膜透析液 细胞外基质 

分 类 号：R285.5[医药卫生—中药学]