灭幽汤对HP相关性胃炎脾胃湿热证小鼠模型中NF-κBp65蛋白,IL-8的表达的影响

Effect of Mieyou Decoction on the Expression of NF-κB p65 Protein and IL-8 in HP Related Gastritis of Spleen Stomach Damp Heat Syndrome Mouse Model

摘要:

目的: 建立幽门螺旋杆菌(Helicobacter pylori,HP)相关性胃炎脾胃湿热证小鼠模型,探讨其与核因子(NF)-κB p65蛋白表达及下游的炎症因子白介素-8(IL-8)相关性,及灭幽汤的作用. 方法: 60只8周龄BALB/c小鼠,雌雄各半,按体重分层随机分为6组:正常组、模型组、灭幽汤组低、中、高剂量(6.2,12.4,24.8 g·kg^-1)、胃三联组(280 mg·kg^-1),对实验组小鼠进行HP相关性胃炎脾胃湿热证造模:适应性饲养3 d予高脂饲养,第11天放入湿热箱中,第16天接种HP,隔日灌胃1次,共3次,定植2周后以胶体金法检测血清HP抗体,第36天药物组予相应浓度药物灌胃,每天1次,连续14 d,其他组正常饲养,第53天处死所有小鼠.观察小鼠一般情况及进食量、饮水量、体重、肛温等变化,光镜下HE染色观察胃黏膜情况并进行积分,免疫组化法检测胃组织中NF-κB p65蛋白表达,酶联免疫吸附法(ELISA)检测血清中IL-8. 结果: 与对照组比较,模型组小鼠胃窦部胃黏膜急慢性炎症积分,NF-κB p65蛋白,IL-8升高(P<0.05);与模型组比较,中、高剂量灭幽汤组及胃三联组的胃窦部胃黏膜急慢性炎症积分,NF-κB p65蛋白,IL-8降低(P<0.05),而低剂量灭幽汤组的变化无统计学差异. 结论: HP相关性胃炎脾胃湿热证发生的机制可能与NF-κB p65蛋白过度表达及IL-8升高而引发炎症反应相关;灭幽汤可能通过抑制NF-κB p65蛋白表达及IL-8而发挥其抗炎作用.

Abstract:

Objective: To study effect of Mieyou decoction on the expression of nuclear factor-κB(NF-κB) p65 protein and IL-8 in Helicobacter pylori(HP) related gastritis of spleen stomach damp heat syndrome mouse model. Method: Sixty BALB/c mice, half male and half female, were randomly divided into 6 groups:control group, model group, Mieyou decoction low, medium, high dose groups(6.2,12.4,24.8 g·kg^-1), gastric triad group(0.28 g·kg^-1). The model was established by high fat feeding and heat box in eleventh days, on sixteenth days with HP was inoculated, and serum HP antibody was detected using colloidal gold method,on thirty-sixth days corresponding drugs were given for 14 days, the other groups received normal feeding. All the mice were sacrificed on fifty-third days. The general condition of mice was observed, and the food and water intake, body weight, body temperature changes were obsedved. HE staining was used to observe the gastric mucosa and integral opticallens, the expression of NF-κB p65 protein was assayed by immunohistochmistry serum IL-8 was determined by ELISA. Result: Compared with control group, the model group increased antral gastric mucosa of acute and chronic inflammation score and expression of NF-κB p65 protein and IL-8 (P<0.05);compared with the model group,the Mieyou decoction middle and high dose group decreased gastric triad antral gastric mucosa of acute and chronic inflammation score and expression of NF-κB p65 protein and IL-8(P<0.05). Conclusion: Mieyou decoction may exert its anti-inflammatory effects through inhibition of NF-κB p65 protein expression and IL-8.