毛樱桃总黄酮对LPS诱导大鼠滑膜细胞RSC-364炎症及自噬的影响

The effect of Prunus tomentosa Thumb total flavones on the inflammation and autophagy of LPS-stimulated rat synovial cells RSC-364

目的探索毛樱桃总黄酮(Prunus tomentosaThumb total flavones,PTTTF)对大鼠滑膜细胞RSC-364炎症反应的影响及其自噬相关机制。方法选用RSC-364细胞株,通过脂多糖(LPS)诱导大鼠滑膜细胞RSC-364建立类风湿关节炎滑膜炎性细胞模型,将细胞分为空白组(CON)、模型组(MOD)、地塞米松(DXM)和不同剂量(0.4、4.0和40.0 mg/L)的PTTTF组。用RT-PCR法检测大鼠滑膜细胞RSC-364细胞中环氧酶1(COX-1)、环氧酶2(COX-2)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、白细胞介素-17(IL-17)及肿瘤坏死因子(TNF-α)表达水平;采用Western blotting法检测大鼠滑膜细胞RSC-364细胞中自噬相关蛋白ATG5、ATG7、ATG12、Beclin1、Lc3II及Bcl-2的表达水平。结果与空白组相比,模型组中COX-2、IL-1β、IL-6、IL-17和TNF-αmRNA表达水平明显升高(P<0.05或P<0.01),自噬相关蛋白ATG5、ATG7、ATG12、Beclin1、Lc3II和Bcl-2蛋白表达水平升高(P<0.05或P<0.01),与模型组相比,PTTTF(0.4、4.0和40.0 mg/L)可明显降低大鼠滑膜细胞RSC-364细胞中炎症因子COX-2、IL-1β、IL-6、IL-17和TNF-αm RNA表达水平(P<0.05或P<0.01);PTTTF(0.4、4.0和40.0 mg/L)可明显抑制大鼠滑膜细胞RSC-364细胞中自噬相关蛋白的表达(P<0.05或P<0.01)。结论 PTTTF抑制大鼠滑膜细胞RSC-364炎症因子的表达及其作用机制可能与抑制自噬相关蛋白表达有关。

This study was performed to explore the effects of Prunus tomentosa Thumb total flavones(PTTTF)on the inflammatory response and autophagy of RSC-364 in rat synovial cells. RSC-364 cell line was selected and induced by lipopolysaccharide(LPS) to construct rheumatoid arthritis synovial inflammatory cell model. RSC-364 cells were divided into blank group(CON), model group(MOD), dexamethasone(DXM) and different doses(0.4, 4.0 and 40.0 mg/L) of the PTTTF group. RT-PCR was used to detect the mRNA expression levels of COX-1, COX-2,IL-1β, IL-6, IL-17 and TNF-α, Western blot was used to detect the expression levels of ATG5, ATG7, ATG12,Beclin1, Lc3 II and Bcl-2. Compared with the blank group, the expression levels of COX-2, IL-1β, IL-6, IL-17 and TNF-α mRNA in the model group were significantly increased(P<0.05 or P<0.01), the expression level of autophagy-related proteins ATG5, ATG7, ATG12, Beclin1, Lc3 II and Bcl-2 were increased(P<0.05 or P<0.01).Compared with the model group, PTTTF(0.4, 4.0 and 40.0 mg/L) significantly decreased the expression levels of inflammatory factors COX-2, IL-1β, IL-6, IL-17 and TNF-α mRNA in RSC-364 cells(P<0.05 or P<0.01),and significantly inhibited the expression of autophagyrelated proteins(P<0.05 or P<0.01). In conclusion,PTTTF can inhibit the expression of RSC-364 inflammatory cytokines in rat synovial cells, and its mechanism may be related to the inhibition of autophagy-related proteins expression.