基于PI3K/AKT信号通路探讨补肾壮骨汤对骨关节炎防治作用的实验研究  

Experimental study on the preventive and therapeutic effects of Bushen Zhuanggu decoction on osteoarthritis based on the PI3K/AKT signaling pathway

作　　者：宋晨阳 孟庆良 SONG Chenyang;MENG Qingliang(Academy of Orthopedics and Traumatology,Henan University of Chinese Medicine,Zhengzhou 450002,China)

机构地区：[1]河南中医药大学骨伤学院,河南郑州450002

出　　处：《陕西中医》2024年第10期1311-1314,1319,共5页Shaanxi Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(81874456);河南省中医药科学研究专项课题(20-21ZYZD05,2019ZY1018,2019ZY2032)。

摘　　要：目的:基于PI3K/AKT信号通路探讨补肾壮骨汤对骨关节炎防治作用的机制。方法:取雄性SD大鼠30只,随机分为对照组、模型组和补肾壮骨组。其中对照组和模型组均给予等体积0.9%氯化钠溶液灌胃;而补肾壮骨组给予补肾壮骨汤灌胃,连续给药4周。干预完成后处死大鼠,采集血液并取关节软骨组织。采用酶联免疫吸附测定(ELISA)检测血清中白细胞介素(IL)-1β、IL-6和IL-10的含量。采用实时定量聚合酶链反应(RT-qPCR)检测磷脂酰肌醇3激酶(PI3K)、丝氨酸-苏氨酸激酶(AKT)和哺乳动物雷帕霉素靶蛋白(mTOR)基因表达;采用Western blot检测抗凋亡基因B淋巴细胞瘤-2(Bcl-2)和Survivin蛋白、促凋亡基因Bcl-2相关X蛋白(Bax)表达。结果:与对照组比较,模型组的软骨细胞活性明显降低;IL-1β和IL-6的含量均明显升高,而IL-10的含量显著下降;PI3K、AKT和mTOR基因和蛋白表达水平均显著降低;Bcl-2和Survivin蛋白表达增加,Bax蛋白表达降低(均P<0.05);而与模型组比较,补肾壮骨组的软骨细胞活性明显增加;IL-1β和IL-6的含量明显下降,而IL-10的含量显著上升;PI3K、AKT和mTOR基因和蛋白表达水平均显著升高;Bcl-2和Survivin蛋白表达下降,Bax蛋白表达上升(均P<0.05)。结论:补肾壮骨汤有助于促进OA软骨细胞增殖、抑制软骨细胞凋亡,发挥抗炎作用,其机制可能与调控PI3K/AKT通路,进而介导软骨细胞增殖凋亡和抗炎作用有关。Objective:To explore the on the preventive and therapeutic effects of Bushen Zhuanggu decoction on osteoarthritis based on the PI3K/AKT signaling pathway.Methods:A total of 30 male SD rats were randomly divided into a control group,a model group and a Bushen Zhuanggu group.Both the control group and the model group were given equal volumes of physiological saline by gavage.The Bushen Zhuanggu group was given Bushen Zhuanggu decoction by gavage for 4 consecutive weeks.After intervention,the rats were euthanized,blood was collected,and articular cartilage tissue was taken.Enzyme linked immunosorbent assay(ELISA)was used to detect the levels of interleukin(IL)-1β,IL-6,and IL-10 in rat serum.Real-time quantitative polymerase chain reaction(RT-qPCR)was applied to detect the gene expressions of phosphatidylinositol 3-kimase(PI3K),v-akt murine thymoma viral oncogene homolog(AKT),and mammalian target of rapamycin(mTOR);while Western blot was employed to detect the protein expressions of PI3K,AKT and mTOR,as well as antiapoptotic gene B lymphoblastoma-2(Bcl-2),Survivin,and proapoptotic gene Bcl-2-associated X protein(Bax).Results:Compared with the control group,the model group had significantly increased chondrocyte activity,obviously increased IL-1βand IL-6 contents,but remarkably decreased IL-10 content;as well as evidently downregulated gene and protein expressions of PI3K,AKT and mTOR;as well as increased Bcl-2 and Survivin protein expressions,and decreased bax protein expression(all P<0.05).While compared with the model group,Bushen Zhuanggu group showed significantly increased chondrocyte activity,obviously reduced IL-1βand IL-6 contents,but remarkably elevated IL-10 content;evidently upregulated gene and protein expressions of PI3K,AKT and mTOR;as well as reduced Bcl-2 and Survivin protein expressions,and elevated Bax protein expression(all P<0.05).Conclusion:Bushen Zhuanggu decoction can promote the proliferation but inhibit the apoptosis of chondrocytes in osteoarthritis,and inhibit inflammatory reactions,whic

关 键 词：骨关节炎 补肾壮骨汤 PI3K/AKT信号通路 软骨细胞 增殖和凋亡 抗炎 抗凋亡基因B淋巴细胞瘤-2 

分 类 号：R684.3[医药卫生—骨科学]