Zhang Xiao-Zhe, Zhang Dong, Ruan An-Min, Li Hui-Juan, Chen Pu, Zhou Jun, Ma Yu-Feng, Qi Yin-Ze, Wang Qing-Fu
The Third Affiliated Hospital of Beijing University of Chinese Medicine, Beijing 100029, China.
Zhongguo Gu Shang. 2020 Jul 25;33(7):643-8. doi: 10.12200/j.issn.1003-0034.2020.07.010.
To observe effects of (, TLZT) gel preparation on p53, miR-502-5p, NF-κBp65 in synovial tissue of knee osteoarthritis (KOA), and to explore mechanism of TLZT gel preparation in treating KOA.
Thirthy-six Wistar rats aged 8 weeks and weighed 200 to 220 g (meaned 208 g) were randomly divided into normal group, model group and traditional Chinese medicine (TCM) group, 12 rats in each group. KOA model was established by modified Hulth method. After 4 weeks of modeling, TCM group treated with TLZT gel preparation for external use, 3 times daily for 2 weeks;normal group and model group were fed normally without intervention. After treatment, morphological changes of specimens in each group were observed, changes of miR-502-5p in synovial tissue were detected by qPCR, and contents of p53, NF-κBp65, IL-1β, TNF-α, MMP-13 in synovial tissue were detected by qPCR and Western Blot respectively.
(1)Morphological observation of specimens showed that the articular cartilage in model group was hyaline and uneven, the synovial membranes were hypertrophic and proliferative with a large number of inflammatory cells infiltrating, the joint fluid was thicker in texture;the articular cartilage in TCM group was more transparent and smooth, synovial hyperplasia was mild with a small amount of inflammatory cell infiltration, the texture of articular fluid was clear and sparse. (2) Compared with normal group, content of miR-502-5p of synovial tissue in model and TCM group were increased, mRNA and expression of p53 decreased, expression of NF-κBp65, IL-1β, TNF-α, MMP-13 increased. (3)Compared with model group, content of miR-502-5p in synovial tissue of TCM group decreased (<0.05), mRNA and protein expression of p53 increased (<0.05), mRNA and protein expression of NF-κBp65, IL-1β, TNF-α, MMP-13 decreased (<0.05).
Expression of p53, miR-502 -5p, NF -κBp65 in synovial tissue is closely related to synovial hyperplasia and inflammatory reaction, TLZT gel preparation may reduce proliferation and inflammatory reaction of KOA synovium by regulating the expression of p53, miR- 502-5p, NF-κBp65 in synovial tissues.
观察通络止痛凝胶剂（TLZT）对膝骨关节炎（KOA）滑膜组织中p53、miR-502-5p、NF-κBp65的影响，探讨TLZT凝胶剂治疗KOA的作用机制。
将36只8周龄、体重200～220g（平均208g）的Wistar大鼠随机分为正常组、模型组和中药组，每组12只。采用改良Hulth法建立KOA模型。造模4周后，中药组外用TLZT凝胶剂，每日3次，连续2周；正常组和模型组正常饲养，不做干预。治疗后，观察各组标本的形态学变化，采用qPCR检测滑膜组织中miR-502-5p的变化，采用qPCR和Western Blot分别检测滑膜组织中p53、NF-κBp65、IL-1β、TNF-α、MMP-13的含量。
（1）标本形态学观察显示，模型组关节软骨呈透明状且不平整，滑膜肥厚、增生，有大量炎性细胞浸润，关节液质地黏稠；中药组关节软骨更透明、光滑，滑膜增生较轻，有少量炎性细胞浸润，关节液质地清亮、稀薄。（2）与正常组比较，模型组和中药组滑膜组织中miR-502-5p含量升高，p53的mRNA及表达降低，NF-κBp65、IL-1β、TNF-α、MMP-13表达升高。（3）与模型组比较，中药组滑膜组织中miR-502-5p含量降低（P<0.05），p53的mRNA及蛋白表达升高（P<0.05），NF-κBp65、IL-1β、TNF-α、MMP-13的mRNA及蛋白表达降低（P<0.05）。
滑膜组织中p53、miR-502-5p、NF-κBp65的表达与滑膜增生及炎症反应密切相关，TLZT凝胶剂可能通过调节滑膜组织中p53、miR-502-5p、NF-κBp65的表达，减轻KOA滑膜的增生及炎症反应。
