熊果酸对脂多糖诱导胃癌细胞增殖、炎症反应的影响及其与NLRP3炎症小体的关系

Effects of ursolic acid on lipopolysaccharide-induced proliferation and inflammatory response of gastric cancer cells and its relationship with NLRP3 inflammasome

目的观察熊果酸(UA)对脂多糖(LPS)诱导的胃癌细胞增殖及炎症反应的影响,并探讨该作用是否与NOD样受体热蛋白结构域相关蛋白3(NLRP3)炎症小体有关。方法取人胃癌细胞系HGC-27、SGC-7901、BGC-823、MGC-803、AGS及正常人胃黏膜上皮细胞株GES-1,RT-q PCR法检测各细胞系中NLRP3表达。将NLRP3表达最高的胃癌细胞BGC-823分为空白对照组、LPS组、LPS+MCC950组及LPS+UA组,LPS组加入100 ng/m L的LPS、LPS+MCC950组加入50μmol/L的NLRP3抑制剂MCC950及100 ng/m L的LPS、LPS+UA组加入50μmol/L的UA及100 ng/m L的LPS,空白对照组不做任何处理。采用CCK-8法观察各组细胞增殖能力,Western blotting、RT-q PCR法检测各组细胞NLRP3蛋白和m RNA,RT-q PCR法检测各组细胞炎症因子白细胞介素1β(IL-1β)、IL-6、IL-18、肿瘤坏死因子α(TNF-α)m RNA。结果各时点细胞存活率LPS组>空白对照组>LPS+MCC950组、LPS+UA组,NLRP3蛋白和m RNA表达LPS组>LPS+MCC950组、LPS+UA组>空白对照组,IL-1β、IL-6、IL-18、TNF-αm RNA表达LPS组>LPS+MCC950组、LPS+UA组>空白对照组(P均<0.05)。结论UA可抑制LPS诱导的胃癌细胞BGC-823细胞增殖及炎症反应,其机制可能与抑制NLRP3炎症小体的激活有关。

Objective To observe the effects of ursolic acid(UA)on the proliferation and inflammatory response of gastric cancer cells induced by lipopolysaccharide(LPS),and to explore whether this effect is related to the inflammasome of NOD-like receptor pyrin domain containing 3(NLRP3).Methods Human gastric cancer cell lines HGC-27,SGC-7901,BGC-823,MGC-803,AGS and normal human gastric mucosa epithelial cell line GES-1 were cultured,and the expression of NLRP3 in each cell line was detected by RT-qPCR.The gastric cancer cell line BGC-823 with the highest expression of NLRP3 was divided into blank control group,LPS group,LPS+MCC950 group,and LPS+UA group.Cells in the LPS group were added with 100 ng/mL of LPS,and cells in the LPS+MCC950 group were added with 50μmol/L of NLRP3 inhibitor MCC950 and 100 ng/mL of LPS,cells in the LPS+UA group were added with 50μmol/L UA and 100 ng/mL LPS,and cells in the blank control group were not treated.CCK-8 method was used to observe the cell proliferation ability of each group,Western blotting and RT-qPCR were used to detect the protein and mRNA of NLRP3 in cells of each group;RT-qPCR was used to detect the inflammatory factor interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-18(IL-18),and tumor necrosis factor-α(TNF-α)mRNA.Results Cell survival rate at each time point was in the following order:LPS group>blank control group>LPS+MCC950 group,LPS+UA group;NLRP3 protein and mRNA expression levels were as follows:LPS group>LPS+MCC950 group,LPS+UA group>blank control group,and the IL-1β,IL-6,IL-18,and TNF-αmRNA expression was as follows:LPS group>LPS+MCC950 group,LPS+UA group>blank control group(all P<0.05).Conclusion UA can inhibit LPS-induced proliferation and inflammatory response of gastric cancer BGC-823 cells,which may be related to the inhibition of NLRP3 inflammasome activation.