白果内酯通过NLRP3相关通路发挥对乙醇致大鼠胃溃疡的保护作用

Ginkgolide protects against ethanol-induced gastric ulcer in rats through NLRP3 related pathway

背景白果内酯对胃溃疡胃黏膜损伤有一定的保护作用,但其具体的作用机制尚不清楚.目的探讨白果内脂通过NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)通路改善乙醇诱导胃溃疡的分子机制.方法将60只SD大鼠随机分为空白组(A组,不做处理),模型组(B组,乙醇诱导的急性胃溃疡模型),对照组(C组,20 mg/mL奥美拉唑),低剂量组(D组,1 mg/mL白果内酯)、中剂量组(E组,2.5 mg/mL白果内酯)和高剂量组(F组,5 mg/mL白果内酯),每组10只,比较不同组别大鼠的胃液pH值、胃泌素、胃蛋白酶以及溃疡指数(ulcer index,UI)等,采用Elisa方法检测大鼠腹主动脉血清以及眼眶血清中的NLRP3、半胱氨酸蛋白酶-1(Caspase-1)、IL-18和IL-1β、超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)和还原型谷胱甘肽(reduced glutathione,GSH)含量.采用实时荧光定量PCR、免疫印迹(WB)法以及免疫荧光法检测胃组织中NLRP3相关通路中NLRP3、白介素-18(IL-1β)、IL-1β、Caspase-1和调亡相关斑点样蛋白(apoptosis associated blotch-like protein,ASC)的表达量变化.结果B组UI、胃泌素、总酸度和胃蛋白酶总活性均明显高于A组(P<0.01),C组,E组和F组的UI、总酸度和胃蛋白酶总活性均低于B组(P<0.01).C组,D组,E组和F组大鼠血清中SOD值和GSH值较B组明显升高(P<0.01).B组大鼠MDA值、NLRP3、Caspase-1、IL-18、IL-1β、Caspase-1和ASC蛋白的mRNA水平和蛋白表达量均明显高于A组(P<0.01).C组,D组,E组和F组大鼠胃组织中MDA值、NLRP3、Caspase-1、IL-18、IL-1β、Caspase-1和ASC蛋白的mRNA水平和蛋白表达量均低于B组(P<0.01).结论白果内脂可通过NLRP3通路抗炎机制达到胃溃疡的保护作用.

BACKGROUND Ginkgolide has a protective effect on gastric ulcer gastric mucosa damage,but the exact mechanism involved is still not clear.AIM To investigate the molecular mechanism of ginkgolide for protecting against ethanol-induced gastric ulcer by detecting the expression of proteins related to the nod-like receptor thermal protein domain associated protein 3(NLRP3)pathway.METHODS Sixty SD rats were randomly divided into a blank group(group A,rats with no treatment),a model group(group B,rats treated with ethanol),control group(group C,rats treated with ethanol and 20 mg/mL omeprazole),low-dose ginkgolide group(group D,rats treated with ethanol and 1 mg/mL ginkgolide),medium-dose ginkgolide group(group E,rats treated with ethanol and 2.5 mg/mL ginkgolide),and high-dose ginkgolide group(group F,rats treated with ethanol and 5 mg/mL ginkgolide),with ten rats in each group.Gastric pH,gastrin,pepsin,and ulcer index(UI)of rats in different groups were compared.The contents of NLRP3,caspase-1,IL-18,IL-1β,superoxide dismutase(SOD),malondialdehyde(MDA),and reduced glutathione(GSH)in serum were determined by ELISA.The expression levels of NLRP3,interleukin(IL)-18,IL-1β,caspase-1,and apoptosis associated blotch-like protein(ASC)in gastric tissues were detected by real-time fluorescence quantitative PCR,Western blot(WB),and immunofluorescence assay.RESULTS UI,gastrin,total acidity,and total pepsin activity in group B were significantly higher than those in group A(P<0.01),while UI,total acidity,and total pepsin activity in groups C,E,and F were significantly lower than those in group B(P<0.01).Serum levels of SOD and GSH in groups C,D,E,and F were significantly higher than those in group B(P<0.01).Serum MDA and the mRNA and protein expression of NLRP3,caspase-1,IL-18,IL-1β,and ASC in gastric tissue in group B were significantly higher than those in group A(P<0.01).Serum MDA and the mRNA and protein expression of NLRP3,caspase-1,IL-18,IL-1β,and ASC in gastric tissue of rats in groups C,D,E,and F were significantly lower than those in group B(P<0.01).CONCLUSION Ginkgolide protects against ethanol-induced gastric ulcer through the NLRP3 pathway.