杜仲对胶原蛋白代谢的影响及其在腹股沟疝筋膜薄弱大鼠中的应用  

Effect of Eucommia ulmoides Oliver on collagen metabolism and its ap⁃plication in weak fascia of inguinal hernia in rats

作　　者：姜新林[1] 张驰[1] 祝佩瑶 JIANG Xinlin;ZHANG Chi;ZHU Peiyao(Department of General Surgery,Nantong Hospital of Traditional Chinese Medicine,Nantong 226001,China.)

机构地区：[1]南通市中医院普通外科,江苏南通226001

出　　处：《中国病理生理杂志》2023年第4期714-720,共7页Chinese Journal of Pathophysiology

基　　金：江苏省优势学科建设工程项目(No.YSHL0814-389)。

摘　　要：目的:观察杜仲提取物(Eucommia ulmoides Oliver extract, EUOE)对大鼠腹股沟疝筋膜薄弱的影响,并初步探索其对人皮肤成纤维细胞(human skin fibroblasts, HSFs)胶原蛋白代谢的作用机制。方法:将32只SD大鼠随机分为正常对照组、模型组、低剂量(1 g/kg)EUOE治疗(low-dose EUOE, EUOE-L)组和高剂量(2 g/kg)EUOE治疗(high-dose EUOE, EUOE-H)组,每组8只。建立大鼠腹股沟区筋膜薄弱模型,给予EUOE干预后,通过生物力学实验检测筋膜抗张强度并采用HE染色观察筋膜组织学结构。采用0~2 g/L EUOE体外孵育HSFs,CCK-8实验检测其对细胞活力的影响;随后通过细胞黏附实验、划痕实验、Transwell细胞迁移实验及ELISA实验观察EUOE(1 g/L)对体外培养HSFs增殖、黏附、迁移及Ⅰ型胶原蛋白(collagen type Ⅰ, COL Ⅰ)分泌的影响,同时利用Western blot实验初步分析HSFs及大鼠筋膜组织中COL Ⅰ、COL Ⅲ和基质金属蛋白酶1(matrix metalloproteinase-1, MMP-1)的表达情况,以及RhoA/ROCK信号通路的活性。结果:体内动物实验结果显示,与模型组相比,给予EUOE治疗可显著增强筋膜的抗张强度(P<0.05),并改善筋膜的组织形态。体外细胞实验的结果显示,EUOE对人皮肤成纤维细胞无显著毒性,同时1 g/L的EUOE孵育48 h可显著增强细胞活力并促进细胞黏附及迁移(P<0.05)。此外,EUOE还可促进HSFs分泌COL I(P<0.05),上调HSFs及大鼠筋膜组织中COL Ⅰ和COL Ⅲ的表达,激活RhoA/ROCK信号通路,抑制MMP-1的表达(P<0.05)。结论:EUOE可显著增强腹股沟疝筋膜薄弱模型大鼠筋膜的抗张强度,该作用可能与其激活RhoA/ROCK信号通路,调节胶原蛋白的代谢有关。AIM:To investigate the effect of Eucommia ulmoides Oliver extract(EUOE)on the weak fascia of inguinal hernia in rats,and to explore its mechanism on collagen metabolism in human skin fibroblasts(HSFs).METHODS:Thirty-two SD rats were randomly divided into normal group,model group,low-dose(1 g/kg)EUOE(EUOE-L)group and high-dose(2 g/kg)EUOE(EUOE-H)group,with 8 rats in each group.The model of weak fascia in the inguinal region of rats was established,and then EUOE intervention was performed.The tensile strength of fascia was detected by biomechanical test,and the histological structure of the fascia was observed by HE staining.The HSFs were incubated with 0~2 g/L EUOE,and the viability was assessed with CCK-8 assay.Subsequently,the effects of EUOE(1 g/L)on proliferation,adhesion,migration and collagen type I(COL I)secretion of HSFs in vitro were detected by cell adhesion assay,scratch test,Transwell migration assay and ELISA.Meanwhile,the expression of COL I,COL III and matrix metalloproteinase-1(MMP-1),and the activation of RhoA/ROCK signaling pathway in HSFs and fascia were evaluated by Western blot analysis.RESULTS:In vivo results showed that,compared with model group,EUOE administration significantly increased the tensile strength of fascia(P<0.05),and improved the morphology of the fascia tissue.In vitro results showed that EUOE is non-cytotoxic to human skin fibroblast,and incubation with 1 g/L EUOE for 48 h could promote cell viability,adhesion and migration(P<0.05).Furthermore,EUOE enhanced COL I secretion in HSFs(P<0.05),and up-regulated the expression of COL I and COL III(P<0.05),concomitant with RhoA/ROCK activation(P<0.05),while inhibited MMP-1 expression both in HSFs and fascial tissue(P<0.05).CONCLUSION:EUOE improves the tensile strength of the weak fascia in inguinal hernia in rats,which may be related to RhoA/ROCK activation and regulation of collagen metabolism.

关 键 词：杜仲 腹股沟疝 胶原蛋白代谢 成纤维细胞 RhoA/ROCK信号通路 

分 类 号：R656.21[医药卫生—外科学] R285.5[医药卫生—临床医学]