白术内酯Ⅰ对IL-1β诱导的关节软骨细胞炎性损伤的影响

Effect of atractylenolide I on the inflammatory injury of articular chondrocytes induced by IL-1β

目的探讨白术内酯I(Atractylenolide I,AT-I)对白细胞介素-1β(interleukin-1β,IL-1β)诱导的关节软骨细胞炎性损伤的影响。方法人关节软骨细胞来自武汉普诺赛生命科技有限公司。实验分6组:对照组(未处理)、模型组(10 ng/mL IL-1β)、AT-I-L组(25μmol/L)、AT-I-M组(50μmol/L)、AT-I-H组(100μmol/L)、AT-I-H+740 Y-P组[100μmol/L AT-I与50μg/mL 740 Y-P磷脂酰肌醇3-激酶(PI3K)激活剂]。CCK-8法和流式细胞术分别测定增殖及凋亡;ELISA法测定上清肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)水平;Western blot检测PI3K/蛋白激酶B(AKT)/核因子-κB(NF-κB)相关蛋白表达。结果与对照组相比,模型组OD450值(24、48 h)降低(P<0.05),TNF-α、IL-6表达、凋亡率、PI3K/AKT/NF-κB蛋白表达均升高(P<0.05)。与模型组对比,AT-I-L组OD450值(24、48 h)、TNF-α、IL-6表达、凋亡率、PI3K/AKT/NF-κB通路蛋白表达差异不显著(P>0.05),AT-I-M组、AT-I-H组OD450值(24 h、48 h)升高(P<0.05),TNF-α、IL-6表达、细胞凋亡率、PI3K/AKT/NF-κB通路蛋白表达均降低(P<0.05)。与AT-I-H组对比,AT-I-H+740 Y-P组OD450值(24、48 h)降低(P<0.05),TNF-α、IL-6表达、凋亡率、PI3K/AKT/NF-κB通路蛋白表达均升高(P<0.05)。结论白术内酯Ⅰ改善IL-1β诱导的关节软骨细胞炎性损伤,其可能机制是促进软骨细胞增殖并抑制炎症反应、细胞凋亡和PI3K/AKT/NF-κB通路实现。

Objective To investigate the effect of atractylenolide I(AT-I)on the inflammatory injury of articular chondrocytes induced by interleukin-1β(IL-1β).Methods Human articular chondrocytes were from Wuhan Punosai Life Technology Co.,LTD.The experiment was divided into six groups:control group(untreat),model group(10 ng/mL IL-1β),AT-I-L group(25μmol/L),AT-I-M group(50μmol/L),AT-I-H group(100μmol/L),and AT-I-H+740 Y-P group(100μmol/L AT-I and 50μg/mL 740 Y-P phosphatidylinositol 3-kinase(PI3K)activator).CCK-8 method and flow cytometry were applied to determine the proliferation and apoptosis,respectively.ELISA method was applied to measure the level of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in the supernatant.Western blotting was applied to detect the expression of PI3K/protein Kinase B(AKT)/Nuclear factor-κB(NF-κB)pathway related proteins.Results Compared to those in the control group,OD450 values(24 h,48 h)in the model group reduced(P<0.05),the TNF-α,IL-6 expression,apoptosis rate,and PI3K/AKT/NF-κB pathway protein expression increased obviously(P<0.05).Compared with the model group,the AT-I-L group showed no obvious differences in OD450 values(24 h,48 h),TNF-α,IL-6 expression,apoptosis rate,and PI3K/AKT/NF-κB pathway protein expression(P>0.05).OD450 values(24 h,48 h)in the AT-I-M and AT-I-H groups increased(P<0.05),and TNF-α,IL-6 expression,apoptosis rate,and PI3K/AKT/NF-κB pathway protein expression decreased(P<0.05).Compared to those in the AT-I-H group,OD450 values(24 h,48 h)in the AT-I-H+740 Y-P group reduced(P<0.05),TNF-αand IL-6 expression,cell apoptosis rate,and PI3K/AKT/NF-κB pathway protein expression increased(P<0.05).Conclusion AT-I improves the inflammatory injury of articular chondrocytes induced by IL-1β.The possible mechanism is to promote chondrocyte proliferation and to inhibit inflammatory response,apoptosis,and PI3K/AKT/NF-κB pathway.