健脾解毒方对幽门螺杆菌诱发胃癌过程微血管密度及环氧合酶表达的影响

Effect of Jianpi Jiedu Recipe on Microvessel Density and Cyclooxygenase-2 Expression in Helicobacter pylori Induced Gastric Cancer

目的探讨健脾解毒方在幽门螺杆菌(Helicobacter pylori,H.pylori)长期感染诱发C57BL/6小鼠胃癌过程中对微血管密度(microvessel density,MVD)和环氧合酶-2(COX-2)的调控作用,为其治疗H.pylori相关性胃病提供实验依据。方法用H.pylori标准株悉尼株(H.pyloriSS1)灌胃的方法建立H.pylori感染C57BL/6小鼠诱发胃癌动物模型,分为对照组、模型组、健脾解毒方低剂量组和健脾解毒方高剂量组,每组40只小鼠。给药72周后处死小鼠,免疫组化法检测各组小鼠胃黏膜MVD的变化;实时荧光定量PCR法和免疫组化法检测各组小鼠胃黏膜COX-2mRNA和蛋白的表达。结果对照组、模型组、健脾解毒方低、高剂量组胃癌的发生率分别为0、22.2%、11.1%和10.0%;胃黏膜MVD分别为(2.50±1.54)、(18.56±2.62)、(14.61±3.60)和(7.39±1.75)个/cm2,模型组MVD较对照组明显增高(P<0.01),健脾解毒方低、高剂量组MVD显著降低(P<0.01)。模型组COX-2mRNA和蛋白表达较对照组均明显增加(P<0.01);健脾解毒方低、高剂量组均可降低其表达,并呈现剂量依赖关系。结论 H.pylori感染可增加C57BL/6小鼠胃黏膜MVD,促进COX-2的表达,可能在H.pylori诱导的胃癌发生中起到促进作用;健脾解毒方可减少小鼠胃黏膜MVD,抑制COX-2的表达,这可能是其防治胃癌的重要机制之一。

Objective To investigate the regulatory effect of Jianpi Jiedu Recipe （JJR） on the microvessel density （MVD） and cyclooxygenase-2 （COX-2） in long-term infection of Helicobacter pylori induced gastric cancer of C57BL/6 mice, thus providing experimental bases for its treatment of the H. pyloricorrelated gastropa- thy. Methods C57BL/6 mouse gastric cancer model induced by H. pylori infection was established by gastrogavage of H. pylori standard strain SSI. Mice were divided into the control group, the model group, low dose J JR group, and the high dose JJR group, 40 in each group. Mice were sacrificed after 72-week medication. Changes of the gastric mucosa MVD of mice in each group were detected by immunohistochemical method. Expressions of COX-2 mRNA and protein were detected by Real-time fluorescent quantitative polymerase chain reaction and immunohistochemical method. Results The occurrence rate of gastric cancer in the control group, the model group, the low dose J JR group, and the high dose J JR group was 0, 22.2%, 11.1%, and 10.0%, respectively. The gastric mucosa MVD （ number/cm2） of mice in each group was 2.50 ± 1.54, 18.56±2.62, 14. 61± 3.60, and 7.39 ±1.75, respectively. The gastric mucosa MVD in the model group increased more obviously than that in the control group （P〈0.01）. The gastric mucosa MVD significantly decreased in the low dose JJR group and the high dose JJR group （P〈0.01）. Expressions of COX-2 mRNA and protein in the model group in- creased more obviously than those in the control group （P〈0.01）. Low dose J JR and high dose JJR could decrease their expressions in a dose dependent manner. Conclusions H. pylori infection could increase the gastric mucosa MVD of C57BL/6 mice and promote COX-2 expressions, which might play a promoting effect in the incidence of H. pylori induced gastric cancer. J JR could decrease the gastric mucosa MVD and inhibit COX-2 expressions, which might be one of its important mechanisms of preventing and treating gastric cancer.