摘要
目的从糖酵解探讨滑膜巨噬细胞(synovial macrophage,SM)极化对膝骨关节炎(knee osteoarthritis,KOA)炎症反应的作用及通痹健骨方的干预机制。方法采用改良Hulth法制作KOA大鼠模型,SD大鼠随机分为空白组(Control)、模型组(Model)、糖酵解抑制剂组(Inhibitor)、通痹健骨方低、中和高剂量组(TBJGF-L、TBJGF-M和TBJGF-H),造模成功后各组给予不同干预措施,HE染色观察滑膜炎症,Masson染色观察滑膜纤维化,ELISA检测血清IL-8、IL-10和HK2含量,生化法检测滑膜组织中LA和ATP含量,多重免疫荧光法检测滑膜组织中M1型、M2型巨噬细胞数量以及巨噬细胞内HK2、LC3B的表达水平。结果Model组滑膜组织表面充血、水肿和增厚,滑膜细胞大量增生,排序杂乱,且滑膜细胞迁移至内膜,炎症细胞浸润和血管数量显著增多,滑膜表现大量纤维化;与Control组相比,Model组血清HK2、IL-8、LA、ATP水平升高(P<0.05),IL-10表达水平降低(P<0.05),M2/M1比值均降低(P<0.05),CD86、HK2和LC3B共定位细胞数量均显著升高(P<0.05)。与Model组相比,Inhibitor组、中药各组炎症细胞浸润、血管数量和滑膜纤维化等均有所缓解,HK2、IL-8和LA、ATP表达水平降低(P<0.05),IL-10表达水平升高(P<0.05),M2/M1比值升高(P<0.05),CD86、HK2和LC3B共定位细胞数量均明显降低(P<0.05),以TBJGF-M组差异最明显(P<0.05)。结论HK2/LC3B介导的SM极化失衡加剧KOA滑膜炎症,通痹健骨方能够抑制HK2、LA和ATP的高表达以缓解糖酵解代谢的异常增加。
Objective To explore the effect of synovial macrophage on the intervention of KOA and the intervention mechanism of TongbiJiangu recipe based on glycolysis.Methods KOA model rats were treated with modified Hulth method.SD ratswere divided into 6 subgroups according to the random method:blank group,model group,glycolysis inhibitor group,TongbiJiangu recipe low-dose and medium-dose groups.After successful modeling,each group was treated with different intervention method.HE staining was used to observe synovial inflammation and Masson staining was used to observe synovial fibrosis.The expression levels of IL-8,IL-10 and HK2 were detected by ELISA.The content levels of LA and ATP in synovial tissue were detected by biochemistry.The number of M1 and M2 macrophages in synovial tissue and the expression levels of HK2 and LC3B in macrophages were detected by multiple immunofluorescence.Results The synovial tissue surface of the knee in the model group was congested,edema and thickening,and the synovial cells were proliferated and disordered,and the synovial cells migrated to the intima,the number of inflammatory cells and blood vessels increased significantly,and the synovial tissue showed a lot of fibrosis.Compared with blank group,the expression levels of serum HK2,IL-8,LA and ATP in model group were increased(P<0.05),the expression levels of IL-10 and E2 were decreased(P<0.05),the M2/M1 ratio in synovial tissue of knee joint was significantly decreased(P<0.05),and the number of CD86,HK2 and LC3B colocalized cells in synovial tissue of knee joint was significantly increased(P<0.05).Compared with the model group,the inflammatory cell infiltration,the number of blood vessels and synovial fibrosis in inhibitor group and TCM groups were alleviated,the expression levels of HK2,IL-8,LA and ATP were decreased(P<0.05),the expression levels of IL-10 were increased(P<0.05),the ratio of M2/M1 in synovial tissue of knee joint was significantly increased(P<0.05),the number of CD86,HK2 and LC3B colocalized cells in the synovial tissue of knee joint was significantly decreased(P<0.05),and the expression level of CD86,HK2 and LC3B was most obvious in the TBJGF-M group.Conclusion The imbalance of SM polarization mediated by HK2/LC3B aggravates the synovial inflammation of KOA.TongbiJiangu prescription can inhibit the high expression of HK2,LA and ATP in KOA rats to alleviate the abnormal increase of glycolytic metabolism.
作者
王晓萍
周明旺
何帮靖
吉星
魏长浩
WANG Xiaoping;ZHOU Mingwang;HE Bangjing;JI Xing;WEI Changhao(Gansu traditional Chinese medicine,Lanzhou 730050,China;Gansu Provincial Academy of Chinese Medicine,Lanzhou 730050,China;Gansu University of Chinese Medicine,Lanzhou 730000,China)
出处
《中国骨质疏松杂志》
北大核心
2025年第12期1735-1740,1745,共7页
Chinese Journal of Osteoporosis
基金
中央引导地方科技发展资金项目(24ZYQA037)
甘肃省骨关节退行性疾病临床医学研究中心项目(18JR2FA009,2020-0411-SFC-0002)
甘肃省重点研发计划-社会发展领域项目(22YF7FA103,25YFFA059)。
