摘要
目的探讨加味少腹逐瘀汤通过调控线粒体自噬发挥拮抗子宫内膜异位症纤维化的机制。方法构建动物模型后以一般状况、器官含水率、热成像评价证候;以异位灶重量和粘连程度评价疗效;HE染色、透射电镜、Masson和天狼星红染色比较病理变化;免疫组化检测PINK1、Parkin的表达;qPCR和Western blot测定mRNA和蛋白表达,ELISA检测血清ROS水平。结果模型小鼠自主活动减弱,脏器含水率增加,热成像见四肢和下腹部温度降低;HE染色可见异位上皮及腺体增生明显,透射电镜可见线粒体、内质网结构损伤,正常自噬结构消失;Masson和天狼星红染色显示胶原沉积加重;免疫组化显示异位灶PINK1、Parkin表达降低;qPCR和Western blot显示模型小鼠异位灶PINK1、Parkin、Beclin1、LC3 mRNA及蛋白表达降低,p62 mRNA及蛋白表达增加,血清ROS升高。加味少腹逐瘀汤干预后模型小鼠证候表现改善;异位灶炎症浸润缓解,线粒体和内质网形态恢复,出现正常自噬结构;胶原沉积及纤维化程度减轻;PINK1、Parkin、Beclin1、LC3 mRNA及蛋白表达增加;p62 mRNA及蛋白表达降低,ROS水平降低。结论加味少腹逐瘀汤可以改善寒湿瘀结证子宫内膜异位症小鼠异位病灶纤维化,可能与调控线粒体自噬有关。
Aim To explore the mechanism of Jiawei Shaofu Zhuyu decoction in antagonizing endometriosis fibrosis by regulating mitophagy.Methods After the animal model was constructed,the syndrome was evaluated by general condition,organ water content and thermal imaging.The curative effect was evaluated by the weight of ectopic focus and the degree of adhesion.The pathological changes were compared using HE staining,transmission electron microscopy,Masson and Sirius red staining.The expression of PINK1 and Parkin was detected by immunohistochemistry.The expression of mRNA and protein was determined by qPCR and Western blot,and the level of serum ROS was detected by ELISA.Results The autonomic activity of model mice was weakened,the water content of organs rose,and the temperature of limbs and lower abdomen was reduced by thermal imaging.HE staining showed obvious hyperplasia of ectopic epithelium and glands.Transmission electron microscopy showed mitochondrial and endoplasmic reticulum structure damage,and normal autophagy structure disappeared.Masson and Sirius red staining showed increased collagen deposition;immunohistochemistry showed decreased expression of PINK1 and Parkin in ectopic foci.qPCR and Western blot showed that the expression of PINK1,Parkin,Beclin1,LC3 mRNA and protein in ectopic foci of model mice decreased,the expression of p62 mRNA and protein increased,and serum ROS increased.The syndrome performance of model mice was improved after the intervention of Jiawei Shaofu Zhuyu decoction;the inflammatory infiltration of ectopic foci was relieved,the morphology of mitochondria and endoplasmic reticulum was restored,and normal autophagy structure appeared.The degree of collagen deposition and fibrosis was reduced;the mRNA and protein expression of PINK1,Parkin,Beclin1 and LC3 increased.The expression of p62 mRNA and protein decreased,and the level of ROS decreased.Conclusions Jiawei Shaofu Zhuyu decoction can improve the fibrosis of ectopic lesions in mice with endometriosis of cold-dampness stasis syndrome,which may be related to the regulation of mitophagy.
作者
黄灿灿
万文文
吉秀家
岳斌
张育贵
张小花
梁莉
陈国廉
武权生
毛海燕
HUANG Can-can;WAN Wen-wen;JI Xiu-jia;YUE Bin;ZHANG Yu-gui;ZHANG Xiao-hua;LIANG Li;CHEN Guo-lian;WU Quan-sheng;MAO Hai-yan(Clinical School of Chinese Medicine,Gansu University of Chinese Medicine,Lanzhou 730000,China;School of Nursing,Gansu University of Chinese Medicine,Lanzhou 730000,China;Scientific Research and Experimental Center,Gansu University of Chinese Medicine,Lanzhou 730000,China;Gansu Provincial People′s Hospital,Lanzhou 730000,China)
出处
《中国药理学通报》
北大核心
2025年第6期1177-1185,共9页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 82260949,81700879)
甘肃省科技计划项目优秀博士生项目(No 23JRRA1220)
甘肃省科技计划项目自然科学基金(No 23JRRA1757,24JRRA1016)
甘肃省中医药科研课题(No GZKP-2023-21)
甘肃省中医药研究中心开放课题(No zyzx-2023-09)
甘肃省名中医传承工作室建设项目(2022-8)
甘肃省高校中(藏)药化学与质量研究省级重点实验室开放基金项目(No zzy-2022-01)
兰州市科技发展指导性计划项目(No 2023-ZD-237)
甘肃省人民医院内部科研基金项目(No 23GSSYE-1)。
