基于NF-κB和JAK/STAT通路探讨黄芪、当归、忍冬藤水提冻干粉调控大鼠滑膜细胞增殖和凋亡的作用机制

Effect and mechanism of Milkvetch Root, Chinese Angelica and Honeysuckle Stem water-extracted lyophilized powder on proliferation and apoptosis of rats synoviocytes based on NF-κB and JAK/STAT signaling pathway

目的基于NF-κB和JAK/STAT通路,探讨黄芪、当归、忍冬藤水提冻干粉对IL-1β诱导的大鼠滑膜细胞增殖和凋亡的影响及作用机制。方法取生长状态良好的RSC-364细胞,分为5组:正常组无药物干预,模型组加10 ng/mL IL-1β,对照组加10 ng/mL IL-1β、10 ng/mL IL-1RA,100μg/mL药物组加10 ng/mL IL-1β和100μg/mL黄芪、当归、忍冬藤水提冻干粉,250μg/mL药物组加10 ng/mL IL-1β和250μg/mL黄芪、当归、忍冬藤水提冻干粉,分别于培养12 h、24 h后流式细胞术检测各组滑膜细胞凋亡率,HE染色观察滑膜细胞形态,Western-Blot法检测滑膜细胞中NF-κB和JAK/STAT通路关键蛋白NF-κB p50、IKKα/β、JAK1、STAT3表达水平。结果模型组培养12 h和24 h细胞凋亡率均显著低于正常组(P均<0.05),250μg/mL药物组培养12 h后细胞凋亡率显著高于模型组(P<0.05)。模型组培养12 h和24 h后细胞中TRAF2、IKKα/β、NF-κB p50、JAK1、STAT3表达水平均明显高于正常组(P均<0.05),而250μg/mL药物组均明显低于模型组(P均<0.05);100μg/mL药物组STAT3表达水平与模型组比较差异无统计学意义(P均>0.05),TRAF2、IKKα/β、NF-κB p50、JAK1表达水平也均明显低于模型组(P均<0.05)。结论黄芪、当归、忍冬藤水提冻干粉可能通过抑制NF-κB和JAK/STAT信号通路促进滑膜细胞凋亡,抑制IL-1β诱导的滑膜细胞过度增殖,减轻炎症反应。

Objective It is to observe the effect and mechanism of Milkvetch Root, Chinese Angelica and Honeysuckle Stem water-extracted lyophilized powder on Il-1β-induced proliferation and apoptosis of rats synoviocytes based on NF-κB and JAK/STAT signaling pathway. Methods RSC-364 cells with good growth status were selected and divided into 5 groups: normal group without drug intervention, model group treated with 10 ng/mL IL-1β, control group treated with 10 ng/mL IL-1βand 10 ng/mL IL-1 RA, 100 μg/mL drug group treated with 10 ng/mL IL-1β and 100 μg/mL astragalus, angelica, honeysuckle water-extracted lyophilized powder, 250 μg/mL drug group was treated with 10 ng/mL IL-1β and 250 μg/mL astragalus, angelica, honeysuckle water-extracted lyophilized powder. The apoptosis rate of synovial cells in each group was detected by flow cytometry after 12 h and 24 h of culture. The morphology of synovial cells was observed by HE staining. The expression levels of NF-κB and JAK/STAT key proteins NF-κB p50, IKKα/β, JAK1 and STAT3 in synovial cells were detected by Western-Blot method. Results The apoptotic rate after 12 h and 24 h of culture in the model group was significantly lower than that in the normal group(P<0.05), and the apoptotic rate after 12 h of culture in 250 μg/mL drug group was significantly higher than that in the model group(P<0.05). The expression levels of TRAF2, IKKα/β, NF-κB p50, JAK1, and STAT3 in the model group were significantly higher than those in the normal group after 12 and 24 hours of culture(P<0.05), and the expression levels in 250 μg/mL drug group were significantly lower than those in the model group(P<0.05). The expression level of STAT3 in the 100 μg/mL drug group was not significantly different from the model group(P>0.05), and the expression levels of TRAF2, IKKα/β, NF-κB p50, and JAK1 were also significantly lower than those in the model group(P<0.05). Conclusion Astragalus membranaceus, Angelica sinensis, and Honeysuckle water-extracted lyophilized powder can promote the apoptosis of synovial cells, inhibit the proliferation of synovial cells induced by IL-1β, and relieve the inflammatory response by inhibiting the NF-κB and JAK/STAT signal pathways.