海南医学院学报2024，Vol.30Issue(22)：1708-1716,9.DOI:10.13210/j.cnki.jhmu.20240710.001
化瘀祛湿方对miR-21靶向Skp2介导早期膝骨关节炎软骨细胞自噬的影响
Effect of Huayu Qushi Decotion on miR-21-targeted Skp2-mediated autophagy of early knee osteoarthritis
摘要
Abstract
Objective:To study the effect of miR-21-5p mediated Skp2 on autophagy of knee OA chondrocytes,and to ex-plore the protective effect of Huayu Qushi Decotion on OA chondrocytes.Methods:(1)Human knee OA cartilage tissues from to-tal knee arthroplasty were taken,and miR-21-5p expression and localization were detected by FISH single-staining assay,Skp2 expression by immunofluorescence,apoptosis by TUNEL staining assay,and relative expressions of Bax,Bcl-2,Beclin1,CARM1,LC3Ⅱ/Ⅰ and Skp2 were determined by Western blot analysis;(2)A total of 36 SD rats were divided into 6 groups ac-cording to the randomized numerical method with 6 rats in each group,including the normal control group,the sham operation group,the model group,the model+airborne group,the model+miR-21-5p interference group,the model+Huayu Qushi Decotion group;the morphology and structure of cartilage were observed by paraffin section alisin blue staining,and the cartilage was detect-ed by qPCR and Western blot method in each group.The relative expressions of Beclin1,CARM1,LC3Ⅱ/Ⅰ and Skp2 were de-tected by qPCR and Western blot,and the IOD/Area value of Skp2 was detected by immunohistochemistry.Results:FISH assay showed that miR-21-5p was highly expressed in human knee OA cartilage tissues,and immunofluorescence assay indicated that Skp2 was lowly expressed in human knee OA cartilage tissues.TUNEL assay revealed a significant increase in apoptosis in human knee OA cartilage tissues.Western blot assay showed that compared with the normal control group,the relative expressions of Bax and CARM1 in human knee OA cartilage tissues were significantly higher compared with the normal group,while the relative expression of Bcl-2,LC3Ⅱ/Ⅰ,Skp2 were significantly lower,and the relative expressions of Beclin1 were slightly lower(P<0.05).Animal experiments showed that the expressions of Beclin1,CARM1,LC3Ⅱ/Ⅰ were significantly decreased and Skp2 was slightly decreased after treatment with miR-21-5p interference.Immunohistochemical detection revealed that the IOD/Area value of Skp2 was significantly decreased in the model group compared with that of the normal control group(P<0.05);com-pared with the model group,the IOD/Area value of the model+Huayu Qushi Decotion IOD/Area value was significantly higher in the model+Huayu Qushi Decotion compared with the model group(P<0.05).Alisin blue staining of paraffin sections showed that the blue color became lighter in the model+airborne group and the model+miR-21-5p interference group compared with that in the sham operation group,but the blue color became darker in the model group,and the blue color was more pronounced in the model+Huayu Qushi Decotion.Conclusion:miR-21-5p can negatively regulate Skp2,mediate knee OA chondrocyte autophagy,and accelerate the development of OA.The Huayu Qushi Decotion may play a role in delaying the degeneration of knee OA by in-hibiting miR-21-5p and increasing the expression of Skp2.关键词
骨关节炎/miR-21/泛素连接酶S期激酶相关蛋白2/化瘀祛湿方/自噬Key words
Chondrocyte osteoarthritis/MiR-21/Skp2/Huayu Qushi Decotion/Autophagy分类
医药卫生引用本文复制引用
袁长深,李彦宏,刘晋邑,袁景钊,梅其杰,徐文飞,段戡..化瘀祛湿方对miR-21靶向Skp2介导早期膝骨关节炎软骨细胞自噬的影响[J].海南医学院学报,2024,30(22):1708-1716,9.基金项目
This study was supported by National Natural Science Foundation of China(82060875 and 82160912) （82060875 and 82160912）
Guangxi Natural Science Foundation(2023GXNSFAA026051) （2023GXNSFAA026051）
Graduate Education Innovation Program of Guangxi University of Traditional Chinese Medicine(YCSW2024401) 国家自然科学基金资助项目(82060875 （YCSW2024401）
82160912) （）
广西自然科学基金资助项目(2023GXNSFAA026051) （2023GXNSFAA026051）
广西中医药大学研究生教育创新计划项目(YCSW2024401) （YCSW2024401）
