姜黄素对肺炎链球菌诱导BEAS-2B细胞SLPI、TNF-α和IL-1β表达的影响

Effect of curcumin on expression of SLPI,TNF-α and IL-1β in BEAS-2B cells induced by Streptococcus pneumoniae

目的:探讨姜黄素(Cur)及其衍生物Y20和6B对肺炎链球菌(SP)诱导的BEAS-2B细胞分泌型白细胞蛋白酶抑制因子(SLPI)、肿瘤坏死因子α(TNF-α)和白细胞介素1β(IL-1β)表达的调节及可能机制。方法:建立体外SP感染模型,采用q PCR检测对照组、感染组以及Cur、Y20和6B处理组在感染1、3、6和9 h SLPI及感染3、6和9 h TNF-α和IL-1β的mRNA水平,ELISA法检测TNF-α和IL-1β的蛋白分泌水平,Western blot实验检测各时点Toll样受体2(TLR2)和磷酸化核因子κB(NF-κB)p65的蛋白水平。结果:q PCR结果显示,Cur、Y20和6B药物处理组SLPI的mRNA表达较SP感染组增加(P<0.05),TNF-α和IL-1β的mRNA表达较SP感染组明显下降(P<0.05);ELISA结果显示,感染不同时点的TNF-α和IL-1β含量在药物处理组中均下降(P<0.05);Western blot结果显示,SP感染3、6和9 h在药物处理组中TLR2和p-NF-κB p65的蛋白水平较感染组下降(P<0.05)。结论:在不同感染时间,Cur、Y20和6B促进SP感染的BEAS-2B细胞SLPI表达,减少TNF-α和IL-1β分泌,其机制可能与抑制TLR2表达、下调NF-κB转录活性有关。

AIM： To explore the effect of curcumin（ Cur） and curcuminoids（ Y20 and 6 B） on the expression of secretory leukocyte protease inhibitor（ SLPI）,tumor necrosis factor-α（ TNF-α） and interleukin-1β（ IL-1β） induced by Streptococcus pneumoniae（ SP） and the possible mechanism. METHODS： BEAS-2 B cells incubated with SP were set up as an inflammation model of pneumonia. The mRNA levels of SLPI at 1 h,3 h,6 h and 9 h,and the mRNA expression of TNF-α and IL-1β at 3 h,6 h and 9 h in control group,SP infection group,Cur treatment group,Y20 treatment group and 6 B treatment group were measured by q PCR. The protein levels of TNF-α and IL-1β in the culture supernatant were measured by ELISA. The protein levels of Toll-like receptor 2（ TLR2） and phosphorylated nuclear factor-κB（ p-NF-κB）p65 at 3 h,6 h and 9 h were determined by Western blot. RESULTS： The mRNA level of SLPI was increased in Cur,Y20 and 6 B treatment groups compared with SP group（ P 〈 0. 05）. The protein levels of TLR2 and p-NF-κB p65 were significantly increased after SP stimulation. After treatment with Cur,Y20 and 6 B,the protein levels of TLR2 and p-NF-κB p65 were significantly decreased（ P 〈 0. 05）. The levels of TNF-α and IL-1β were significantly increased after SP stimulation. Cur,Y20 and 6 B significantly decreased the levels of TNF-α and IL-1β in the supernatant（ P 〈 0. 05）. CONCLUSION： Cur,Y20 and 6 B increase SLPI expression,reduce the expression of inflammatory cytokines TNF-α and IL-1β.The possible mechanism might be associated with inhibiting TLR2 expression and down-regulating the transcriptional activity of NF-κB.