由粪便代谢组学研究热敏灸改善膝骨性关节炎的机制

Study on the mechanism of heat-sensitive moxibustion in improving knee osteoarthritis based by fecal metabolomics

目的采用粪便代谢组学技术,探讨热敏灸治疗膝骨性关节炎的潜在作用机制。方法复制膝骨性关节炎模型后,将模型组兔随机分为模型组与艾灸组,同时设空白组。分别按各被试因素进行干预14 d后,根据终温变化统计,将艾灸组兔分为热敏灸组(>1℃)和非热敏艾灸组(≤1℃)。应用HE染色观察软骨组织,ELISA检测血清炎症因子含量,LC-MS检测粪便代谢物变化,多元统计分析鉴别差异代谢物并分析代谢通路。结果经HE染色,模型组软骨组织较空白组改变发生显著病理学变化。造模后,模型组IL-1β、IL-6含量较空白组显著升高(P<0.001),IL-10下降显著(P<0.001)。治疗后,相比模型组,热敏灸组、非热敏灸组IL-1β、IL-6含量均显著下降(P<0.001),IL-10含量均上调显著(P<0.001),而热敏灸组IL-1β、IL-6下降趋势更好。经分析,确定10个生物标志物和11条代谢通路与KOA密切相关,如类固醇降解、卵巢类固醇生成、视黄醇代谢等,其中以类固醇降解最为显著。结论热敏灸改善KOA炎症反应,发挥抗KOA作用可能与调控类固醇降解、卵巢类固醇生成、视黄醇代谢等有关。

Objective Fecal metabonomics technique was used to investigate the potential mechanism of heat-sensitive moxibustion to treatment of knee osteoarthritis.Methods After replicating the model of knee osteoarthritis,the rabbits in the model group were randomly divided into model group and moxibustion group.At the same time,set up a blank group.After 14 days of intervention according to the subjects'factors,the rabbits in the moxibustion group were divided into heat-sensitive moxibustion group(>1℃)and non-heat-sensitive moxibustion group(≤1℃)according to the statistics of final temperature changes.The cartilage tissue was observed by HE staining,serum inflammatory factors were detected by ELISA,and fecal metabolites were detected by LC-MS.By multivariate statistical analysis,differential metabolites were screened and metabolic pathways were analyzed.Results After HE staining,the cartilage tissue of model group showed significant pathological changes compared with that of blank group.After the establishment of the model,the contents of IL-1βand IL-6 in the model group were significantly higher than those in the blank group(P<0.001),while IL-10 decreased significantly(P<0.001).After treatment,compared with the model group,the contents of IL-1βand IL-6 in the heat sensitive moxibustion group and non-heat sensitive moxibustion group decreased significantly(P<0.001),and the content of IL-10 increased significantly(P<0.001),while the decreasing trend of IL-1βand IL-6 in the heat sensitive moxibustion group was better.10 biomarkers and 11 metabolic pathways were determined to be closely related to KOA,such as steroid degradation,ovarian steroidogenesis,retinol metabolism and so on,among which steroid degradation was the most significant.Conclusion Heat-sensitive moxibustion being improved the inflammatory reaction of KOA and exert the effect of anti-KOA,which may be related to the regulation of steroid degradation,ovarian steroidogenesis and retinol metabolism.