目的:评估芍药苷通过信号转导与转录激活子3(signal transducer and activator of transcriptions 3，STAT3)对白细胞介素-13(interleukin-13，IL-13)诱导的BEAS-2B哮喘细胞模型的炎症、氧化应激和自噬的改善作用。方法:用1、10和30μmol·L-1的芍药苷干预IL-13诱导前后的BEAS-2B细胞，CCK-8检测细胞增殖活力，筛选芍药苷最佳作用条件。然后利用芍药苷和STAT3激活剂干预IL-13诱导的细胞模型，将细胞分为对照组、模型组、STAT3激活剂组、芍药苷组和STAT3激活剂+芍药苷共处理组。CCK-8检测各组细胞增殖活力，ELISA试剂盒检测各组细胞上清中IL-4、IFN-γ的含量，及细胞中MDA、CAT、SOD水平，Western blot检测各组细胞LC3Ⅱ、LC3 Ⅰ、P62、p-STAT3和STAT3的蛋白表达。结果:CCK-8检测结果显示，10 μmol·L-1的芍药苷作用24 h为最佳干预条件。与对照组相比，模型组细胞增殖能力下降(P＜0。05)，IFN-γSOD和CAT含量下降(P＜0。05)，IL-4和MDA含量增加(P＜0。05)，LC3Ⅱ/Ⅰ蛋白表达比值和p-STAT3/STAT3蛋白表达比值增加(P＜0。05)，P62蛋白表达下降(P＜0。05)。与模型组相比，STAT3激动剂组细胞增殖能力下降(P＜0。05)，IFN-γ、SOD和CAT含量下降(P＜0。05)，IL-4和MDA含量增加(P＜0。05)，LC3Ⅱ/Ⅰ蛋白表达比值和p-STAT3/STAT3蛋白表达比值增加(P＜0。05)，P62蛋白表达下降(P＜0。05);芍药苷组细胞增殖能力增加(P＜0。05)，IFN-γ、SOD和CAT含量增加(P＜0。05)，IL-4和MDA含量下降(P＜0。05)，LC3 Ⅱ/Ⅰ蛋白表达比值和p-STAT3/STAT3蛋白表达比值下降(P＜0。05)，P62蛋白表达增加(P＜0。05)。与STAT3激动剂组相比，STAT3激动剂+芍药苷组共处理逆转了 STAT3激动剂的作用。结论:芍药苷通过抑制STAT3蛋白磷酸化，抑制炎症反应，从而改善哮喘中细胞氧化应激和自噬。
Paeoniflorin inhibits IL-13-induced oxidative stress and autophagy in BEAS-2B cells via STAT3
OBJECTIVE To evaluate the effects of paeoniflorin on interleukin-13(IL-13)-induced inflammation,oxidative stress and autophagy in BEAS-2B asthma cell model via signal transducer and activator of transcriptions 3(STAT3).METHODS BEAS-2B cells were intervened with paeoniflorin(1,10,30 μmol·L-1)before and after IL-13 induction.Cell pro-liferation activity was detected by CCK-8 and optimal conditions of paeoniflorin were screened.Then paeoniflorin and STAT3 activators were utilized for intervening IL-13-induced cell models.The cells were assigned into five groups of control,model,STAT3 activator,paeoniflorin and STAT3 activator+paeoniflorin co-treatment.Cell proliferation activity was detected by CCK-8;IL-4 and IFN-γ contents in supernatant of each group were detected by enzyme-linked immunosorbent assay(ELISA);MDA,CAT and SOD levels were detected by ELISA.And the protein expressions of LC3 Ⅱ,LC3 Ⅰ,P62,p-STAT3 and STAT3 in each group were detected by Western blot.RESULTS The results of CCK-8 indicated that 10 μmol·L-1 of paeoniflo-rin for 24 h was the optimal intervention.As compared with control group,cell proliferation capacity dropped(P＜0.01),the con-tents of IFN-γ,SOD and CAT declined(P＜0.01),the contents of IL-4 and MDA spiked(P＜0.01),the ratio of LC3 Ⅱ/Ⅰprotein expression and the ratio of p-STAT3/STAT3 protein expression rose(P＜0.05),and the expression of P62 protein decreased in model group(P＜0.01).Compared with model group,proliferation capability of cells(P＜0.01)and contents of IFN-γ,SOD and CAT decreased(P＜0.01)while the contents of IL-4 and MDA spiked in STAT3 agonist group(P＜0.01).LC3Ⅱ/Ⅰ and p-STAT3/STAT3 protein expression ratios rose(P＜0.05)while there was a down-regulation of P62 protein expression(P＜0.01).In paeoniflorin group,cell proliferation capacity spiked(P＜0.01),IFN-γ,SOD and CAT contents jumped(P＜0.01),IL-4 and MDA contents dropped(P＜0.01)and LC3 Ⅱ/Ⅰ and p-STAT3/STAT3 protein expression ratios decreased(P＜0.01),and the expression of P62 protein became up-regulated(P＜0.01).As compared with STAT3 ago-nist group,the effect of STAT3 agonist was reversed after a co-treatment of STAT3 agonist and paeoniflorin.CONCLUSION Paeoniflorin may suppress inflammatory responses through blunting the phosphorylation of STAT3 protein and improve oxidative stress and autophagy of asthma cell.
asthmapaeoniflorinsignal transducer and activator of transcriptions 3oxidative stressautophagy
NETL
NSTL
万方数据
