基于SOX9/NF-κB/MMP-13信号通路探讨补肾痹通方含药血清对IL-1β诱导软骨细胞凋亡的影响OACSTPCD
Exploring the effect of Bushen Bitong recipe-containing serum on IL-1β-induced chondrocyte apoptosis based on SOX9/NF-κB/MMP-13 signaling pathway
目的:观察补肾痹通方(BSBT)含药血清对IL-1β诱导软骨细胞凋亡的影响及可能作用机理.方法:第3代大鼠软骨细胞随机分为Control组、IL-1β组、IL-1β+BSBT(L)组、IL-1β+BSBT(M)组、IL-1β +BSBT(H)组(5%、10%、15%BSBT含药血清),分别干预24 h后采用CCK8法、流式细胞术分别检测细胞增殖情况和凋亡率;Western blot检测软骨细胞中B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(BAX)、半胱氨酸天冬氨酸蛋白酶3(Caspase-3)、性别决定区Y框蛋白9(SOX9)、磷酸化细胞核因子(NF-κB p65)、基质金属蛋白酶13(MMP-13)蛋白的表达水平;ELISA法检测软骨细胞培养上清中肿瘤坏死因子-α(TNF-α)、白细胞介素6(IL-6)和碱性成纤维细胞生长因子(bFGF)水平.结果:与Control组比较,IL-1β组细胞增殖活性下降,细胞凋亡率上升,NF-κB p65、MMP-13蛋白水平和TNF-α、IL-6水平升高,SOX9蛋白水平和bFGF水平降低;与IL-1β组比较,不同浓度的补肾痹通方含药血清组细胞增殖活性上升,细胞凋亡率降低,NF-κB p65、MMP-13蛋白水平和TNF-α、IL-6水平降低,SOX9蛋白水平和bFGF水平升高;与IL-1β+BSBT(L)组比较,IL-1β+BSBT(M)组和IL-1β+BSBT(H)组细胞增殖活性上升,细胞凋亡率降低,NF-κB p65、MMP-13蛋白水平和TNF-α、IL-6水平降低,SOX9蛋白水平和bFGF水平升高.结论:补肾痹通方含药血清可能通过SOX9/NF-κB/MMP-13信号通路,促进IL-1β诱导的软骨细胞增殖,减少细胞凋亡,改善软骨细胞微环境,促进软骨修复.
Objective:To observe the effect and possible mechanism of action of Bushen Bitong recipe(BSBT)containing serum on IL-1β-induced chondrocyte apoptosis.Methods:Chondrocytes of the 3rd generation rat were randomized into Control,IL-1β,IL-1β+BSBT(L),IL-1β+BSBT(M),and IL-1β+BSBT(H)groups(5%,10%and 15%BSBT-containing se-rum).The cell proliferation and apoptosis rate were tested 24 h after intervention;Western blot was used to detected the expres-sion levels of Bcl-2,BAX,Caspase-3,SOX9,NF-κB p65 and MMP-13 proteins in chondrocytes;ELISA was used to detected the levels of TNF-α,IL-6,and bFGF in the supernatants of chondrocyte culture.Results:Compared with Control group,cell proliferation activity decreased,apoptosis rate increased,NF-κB p65,MMP-13 protein level and TNF-α,IL-6 level increased,and SOX9 protein level and bFGF level decreased in IL-1β group;compared with IL-1β group,different concentrations of BSBT-containing serum group,cell proliferation activity increased,and apoptosis rate decreased.NF-κB p65,MMP-13 protein level and TNF-α,IL-6 level decreased,SOX9 protein level and bFGF level increased;compared with IL-1β+BSBT(L)group,cell pro-liferation activity increased,apoptosis rate decreased in IL-1β+BSBT(M)and IL-1β+BSBT(H)groups,and NF-κB p65,MMP-13 protein level and TNF-α level and IL-6 levels decreased,and SOX9 protein levels and bFGF levels increased.Conclu-sion:Bushen Bitong recipe-containing serum may promote IL-1β-induced proliferation of chondrocytes,reduce apoptosis,im-prove the microenvironment of chondrocytes,and promote cartilage repair through the SOX9/NF-κB/MMP-13 signaling pathway.
易林;张文豪;向文远;石正誉;热米拉·艾买提;邓迎杰;方锐
新疆医科大学第四临床医学院,新疆 乌鲁木齐 830054||新疆医科大学附属中医医院,新疆 乌鲁木齐 830099新疆医科大学附属中医医院,新疆 乌鲁木齐 830099
中医学
补肾痹通方骨关节炎软骨细胞信号通路IL-1β
Bushen Bitong recipeOsteoarthritisChondrocytesSignaling pathwayIL-1β
《海南医学院学报》 2024 (004)
241-247 / 7
This study was supported by National Natural Science Foundation of China(82360934);Science and Technology Innovation Leading Talents Project of Xinjiang Uygur Autonomous Region(2022TSYCLJ0007);Xinjiang Uygur Autonomous Region Key Research and Development Task Special Project(2021B03006);Natural Science Foundation of Xinjiang Uygur Autonomous Region(2022D01C170;2022D01C171)国家自然科学基金(82360934);新疆维吾尔自治区科技创新领军人才项目(2022TSYCLJ0007);新疆维吾尔自治区重点研发任务专项(2021B03006);新疆维吾尔自治区自然科学基金(2022D01C170;2022D01C171)
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