乌药对大鼠宫腔粘连纤维化的干预作用及其代谢组学分析  

Interventional Effect and Metabolomics Analysis of Linderae Radix on Fibrosis of Rats with Intrauterine Adhesions

作　　者：严婧[1,2] 杨盾 郭东卫 王雅静[1] 刘靓丽 贾天阳 谢明霞 YAN Jing;YANG Dun;GUO Dongwei;WANG Yajing;LIU Liangli;JIA Tianyang;XIE Mingxia(Hunan University of Chinese Medicine,Changsha 410208,China;Yueyang Maternal-Child Medicine Health Hospital,Yueyang 414000,China;Brain Hospital of Hunan Province(The Second People's Hospital of Hunan Province),Changsha 410021,China)

机构地区：[1]湖南中医药大学,长沙410208 [2]岳阳市妇幼保健院,湖南岳阳414000 [3]湖南省脑科医院/湖南省第二人民医院,长沙410021

出　　处：《中国实验方剂学杂志》2023年第15期142-151,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：湖南省自然科学基金项目(2021JJ40551);湖南省中医药管理局科研项目(D2022053);湖南省卫生健康委员会科研计划项目(202107021634,B202313057639);湖南省教育厅一流学科开放基金重点项目(2020ZXYJH12)。

摘　　要：目的:探讨乌药对宫腔粘连(IUA)模型大鼠的药效学影响及其代谢机制。方法:建立了一种基于机械损伤+感染的IUA大鼠模型,通过分子生物学及药理学方法和技术评价乌药提取物(LAE)抑制宫腔粘连纤维化的作用;通过基于气相色谱法-质谱法联用(GC-MS)的血清代谢组学方法探讨LAE的代谢调节机制。结果:动物实验表明,LAE能显著改善IUA模型大鼠子宫组织细胞的形态和结构损伤,子宫内膜增殖、血管生成和形态学恢复,并抑制血清转化生长因子-β_(1)(TGF-β_(1))、Smad2及Smad3 mRNA的表达,而增加Smad7 mRNA表达,以抑制纤维化。此外,LAE可显著抑制雌二醇(E2)、促卵泡生成激素(FSH)和促黄体生成素(LH)等激素水平及肿瘤坏死因子(TNF)-α的表达(P<0.01),改善子宫微环境。代谢组学表明,与正常组相比,IUA大鼠血清代谢发生显著异常,共筛选鉴定得到9个差异标志物。LAE可显著改善这种代谢异常,主要包括6个差异代谢物,其中5个为上述9个标志物中的共有标志物,分别为天门冬氨酸(L-aspartic acid),焦谷氨酸(L-pyroglutamic acid),丝氨酸(L-serine),葡萄糖(Glucose)和正缬氨酸(L-norvaline)。通路富集显示氨酰-tRNA合成通路是其主要的影响机制。结论:结合药理学研究结果可得,LAE可显著改善IUA模型大鼠子宫损伤,抑制纤维化。其机制可能与通过抑制氨酰-tRNA合成通路,改善微环境有关。Objective:To investigate the pharmacological effect and metabolic mechanism of Linderae Radix on the intrauterine adhesion(IUA)rat model.Method:An IUA rat model was induced by mechanical injury and infection.Molecular biology and pharmacology techniques were employed to evaluate the inhibitory effect of Linderae Radix extract(LAE)on fibrosis in IUA.Serum metabolomics analysis based on gas chromatography-mass spectrometry(GC-MS)was conducted to explore the metabolic regulation mechanism of LAE.Result:Animal experiments showed that LAE significantly improved the morphology and structural damage of uterine tissue cells in the IUA rat model,promoted endometrial proliferation,vascular regeneration,and morphological recovery,inhibited the mRNA expression of transforming growth factor-β_(1)(TGF-β_(1)),Smad2,and Smad3,and increased the expression of Smad7 mRNA to suppress fibrosis.Additionally,LAE significantly suppressed the levels of estrogen(E2),follicle-stimulating hormone(FSH),luteinizing hormone(LH),and tumor necrosis factor-α(TNF-α)expression(P<0.01),thereby improving the uterine microenvironment.Metabolomics analysis revealed significant metabolic abnormalities in the serum of IUA rats compared with the results in the normal group,and nine differential metabolites were identified.LAE effectively ameliorated these metabolic abnormalities,primarily by influencing six differential metabolites,including five shared metabolites among the nine identified markers:L-aspartic acid,L-pyroglutamic acid,L-serine,glucose,and L-norvaline.Pathway enrichment analysis indicated that the aminoacyl-tRNA biosynthesis pathway was the main affecting mechanism.Conclusion:In combination with the pharmacological research results,LAE effectively improved uterine damage and inhibited fibrosis in the IUA rat model.Its mechanism may involve the inhibition of the aminoacyl-tRNA biosynthesis pathway and the improvement of the microenvironment.

关 键 词：乌药 宫腔粘连 纤维化 代谢组学 

分 类 号：R284.2[医药卫生—中药学] R285[医药卫生—中医学] R289R287R22R2-031R33R24