摘要
目的:探讨姬松茸多糖(ABP)对白细胞介素1β(IL-1β)诱导软骨细胞CHON-001损伤的保护作用和分子机制。方法:以10 ng·ml^(-1)的IL-1β处理CHON-001细胞建立软骨细胞炎症模型。将CHON-001细胞分为对照(Con)组、IL-1β组、IL-1β+ABP低剂量(ABP-L,10 mg·L^(-1))组、IL-1β+ABP中剂量(ABP-M,20 mg·L^(-1))组、IL-1β+ABP高剂量(ABP-H,40 mg·L^(-1))组、IL-1β+miR-NC组、IL-1β+miR-382-3p组、IL-1β+ABP+anti-miR-NC组和IL-1β+ABP+anti-miR-382-3p组。采用流式细胞术检测细胞凋亡;酶联免疫吸附法检测CHON-001细胞培养液中白细胞介素6(IL-6)、γ干扰素(IFN-γ)和肿瘤坏死因子α(TNF-α)的水平;Western blot检测B细胞淋巴瘤(Bcl-2)和Bcl相关x蛋白(Bax)水平;实时定量PCR检测miR-382-3p表达。结果:与Con组比较,IL-1β组IL-6、IFN-γ、TNF-α水平以及细胞凋亡率、Bax蛋白水平显著升高(P<0.05),miR-382-3p表达及Bcl蛋白水平显著降低(P<0.05)。与IL-1β组比较,IL-1β+ABP-L组、IL-1β+ABP-M组、IL-1β+ABP-H组IL-6、IFN-γ、TNF-α水平以及细胞凋亡率、Bax蛋白水平显著降低(P<0.05),miR-382-3p表达及Bcl蛋白水平显著升高(P<0.05)。与IL-1β+miR-NC组比较,IL-1β+miR-382-3p组IL-6、IFN-γ、TNF-α水平以及细胞凋亡率、Bax蛋白水平显著降低(P<0.05),Bcl蛋白水平显著升高(P<0.05)。与IL-1β+ABP+anti-miR-NC组比较,IL-1β+ABP+anti-miR-382-3p组IL-6、IFN-γ、TNF-α水平以及细胞凋亡率、Bax蛋白水平显著升高(P<0.05),Bcl蛋白水平显著降低(P<0.05)。结论:姬松茸多糖通过上调miR-382-3p表达来抑制IL-1β诱导的软骨细胞炎症反应和凋亡。
Objective:To investigate the protective effects and molecular mechanism of agaricus blazei polysaccharide(ABP) on IL-1β-induced chondrocyte(CHON-001) injury.Methods:CHON-001 cells were treated with 10 ng·ml^(-1)interleukin 1β(IL-1β) to establish a chondrocyte inflammation model.CHON-001 cells were divided into control(Con) group,IL-1β group,IL-1β+ABP low dose(ABP-L,10 mg·L^(-1)) group,IL-1β+ABP medium dose(ABP-M,20 mg·L^(-1)) group,IL-1β+ABP high dose(ABP-H,40 mg·L^(-1)) group,IL-1β+miR-NC group,IL-1β+miR-382-3 p group,IL-1β+ABP+ anti-miR-NC group and IL-1β+ABP+anti-miR-382-3 p group.Flow cytometry was used to detect the apoptosis.Enzyme-linked immunosorbent assay was used to detect the levels of interleukin 6(IL-6),interferon γ(IFN-γ) and tumor necrosis factor α(TNF-α) in CHON-001 cell culture media.Western blot was used to detect the levels of B-cell lymphoma(Bcl-2) and Bcl-related x protein(Bax).Real-time quantitative PCR was used to detect miR-382-3 p expression.Results:Compared with those in the Con group,the IL-6,IFN-γ and TNF-α levels,the apoptosis rate and Bax protein level in IL-1β group were significantly increased(P<0.05),and miR-382-3 p expression and Bcl-2 protein level were significantly decreased(P<0.05).Compared with those in IL-1β group,the IL-6,IFN-γ and TNF-α levels,apoptosis rate and Bax protein level in IL-1β+ABP-L group,IL-1β+ABP-M group and IL-1β+ABP-H group were significantly decreased(P<0.05),and miR-382-3 p expression and Bcl-2 protein level were significantly increased(P<0.05).Compared with those in IL-1β+miR-NC group,the IL-6,IFN-γ and TNF-α levels,apoptosis rate and Bax protein level in IL-1β+miR-382-3 p group were significantly reduced(P<0.05),and Bcl-2 protein level was significantly increased(P<0.05).Compared with those in IL-1β+ ABP+ anti-miR-NC group,the IL-6,IFN-γ and TNF-α levels,apoptosis rate and Bax protein level in IL-1β+ABP+anti-miR-382-3 p group were significantly increased(P<0.05),and Bcl-2 protein level was significantly decreased(P<0.05).Conclusion:Agaricus blazei polysaccharide inhibits IL-1β-induced chondrocytes inflammation and apoptosis by up-regulating miR-382-3 p expression.
作者
蒲超
张珊珊
吴青霞
罗栩伟
王亮
张波
Pu Chao;Zhang Shanshan;Wu Qingxia;Luo Xuwei;Wang Liang;Zhang Bo(Department of Orthopedics,The Second Clinical College of North Sichuan Medical College,Nanchong Central Hospital,Sichuan Nanchong 637000,China;Department of Neurology,The Second Clinical College of North Sichuan Medical College,Nanchong Central Hospital,Sichuan Nanchong 637000,China)
出处
《中国药师》
CAS
2022年第4期573-578,共6页
China Pharmacist
基金
四川省教育厅科研计划(编号:17ZB0176)
南充市市校合作科研项目(编号:19SXHZ0119)。
