鸢尾素通过抑制PI3K/AKT信号通路降低P21的表达缓解软骨细胞衰老延缓小鼠骨关节炎的进展  

Irisin alleviates chondrocyte aging and delays the progression of osteoarthritis in mice by inhibiting PI3K/Akt signaling pathway and reducing P21 expression

作　　者：金叶盛 薛源[2] 张元澍 汪凯 施勤[3] 芮永军[2] Jin Yesheng;Xue Yuan;Zhang Yuanshu;Wang Kail;Shi Qin;Rui Yongjun(Suzhou Medical College of Soochow University,Suzhou 215031,China;Department of Hand Surgery,Wuxi Ninth Hospital Affiliated to Suzhou University,Wuxi 214026,China;Department of Orthopedics,the First Affliated Hospital of Suzhou University,Institute of Orthopedicsof Suzhou University,Suzhou 215006,China)

机构地区：[1]苏州大学苏州医学院,苏州215031 [2]苏州大学附属无锡九院手外科,无锡214026 [3]苏州大学附属第一医院骨科,苏州大学骨科研究所,苏州215006

出　　处：《中华手外科杂志》2022年第6期517-522,共6页Chinese Journal of Hand Surgery

基　　金：无锡市“太湖人才计划”顶级医学专家团队;无锡市卫健委青年项目(Q202130);无锡市卫健委青年项目(Q202046)。

摘　　要：目的按探究鸢尾素对骨关节炎小鼠中衰老因子P21的影响。方法将H_(2)O_(2)和鸢尾素作用于小鼠原代软骨细胞,通过CCK8验证鸢尾素对小鼠原代软骨细胞增殖的影响。通过β半乳糖苷酶染色检测小鼠原代软骨细胞中细胞衰老的情况。通过RT-PCR检测小鼠原代软骨细胞中P21 mRNA的表达情况。通过Westem blot检测小鼠原代软骨细胞中P21蛋白的表达。通过DMM术构造小鼠骨关节炎模型,关节腔注射鸢尾素,通过免疫组化染色和免疫荧光染色,检测小鼠胫骨表面软骨中P21蛋白的表达情况。通过Westerm blot检测PI3K/AKT信号通路相关蛋白的表达。结果CCK8实验结果显示,鸢尾素对小鼠原代软骨细胞的增殖没有显著的影响。β半乳糖苷酶染色结果显示,鸢尾素能改善H_(2)O_(2)对小鼠软骨细胞的衰老作用。RT-PCR的结果表明,鸢尾素能逆转H,O2诱导后小鼠软骨细胞中P21基因的增加。Western blot结果表明,鸢尾素能逆转H,O,诱导后小鼠软骨细胞中P21蛋白的增加。RTPCR的结果表明,内源性缺少鸢尾素会加重H_(2)O_(2)对小鼠软骨细胞中P21基因的诱导作用。Western blot结果表明,内源性缺少鸢尾素会加重H_(2)O_(2)对小鼠软骨细胞中P21蛋白的诱导作用。免疫组化和免疫荧光染色的结果表明,鸢尾素能改善DMM术后骨关节炎小鼠腔骨表面软骨中P21蛋白的表达。Western blot结果显示,鸢尾素能抑制PI3K/AKT信号通路的激活。结论鸢尾素通过抑制PI3K/AKT信号通路降低骨关节炎小鼠软骨细胞中衰老基因P21的表达,延缓骨关节炎的进展。Objective To investigate the effect of irisin on P21 in osteoarthritis mice.Methods H_(2)O_(2) and irisin were applied to mouse primary chondrocytes,and the effect of irisin on the proliferation of mouse primary chondrocytes was verified by CCK8.Theβgalactosidase staining was used to detect cell senescence in mouse primary chondrocytes.The expression of P21 mRNA in mouse primary chondrocytes was detected by RTPCR.The expression of P21 protein in mouse primary chondrocytes was detected by Western blot.A mouse osteoarthritis model was constructed by DMM,irisin was injected into the joint cavity,and the expression of P21 protein in the cartilage of the tibia surface of mice was detected by immunohistochemistry and immunofluorescence staining.The expression of PI3K/Akt signaling pathway related proteins was detected by Western blot.Results The CCK8 test showed that irisin had no significant effect on the proliferation of primary chondrocytes in mice.Theβgalactosidase staining showed that irisin could improve the aging effect of H_(2)O_(2)on mouse chondrocytes.The RT-PCR results showed that irisin could reverse the increase of P21 gene in mouse chondrocytes induced by H_(2)O_(2).The Western blot results showed that irisin could reverse the increase of P21 protein in mouse chondrocytes induced by H_(2)O_(2).The RT-PCR results showed that endogenous lack of irisin would aggravate the induction of P21 gene in mouse chondrocytes by H_(2)O_(2).The Western blot results showed that endogenous lack of irisin would aggravate the induction of P21 protein in mouse chondrocytes by H,O,.The results of immunohistochemistry and immunofluorescence staining showed that irisin could improve the expression of P21 protein in the tibial surface cartilage of osteoarthritis mice after DMM.The Western blot results showed that irisin could inhibit the activation of PI3K/Akt signaling pathway.ConclusionThe irisin can reduce the expression of P21 in chondrocytes of osteoarthritis mice by inhibiting PI3K/Akt signaling pathway,thus delaying the

关 键 词：骨关节炎 细胞衰老 鸢尾素 P21 PI3K/AKT 

分 类 号：R285.5[医药卫生—中药学]