基于ERK/Cyclin E通路探讨健脾化瘀解毒方逆转CAG大鼠胃黏膜癌变效应机制

Exploring Mechanism of Jianpi Huayu Jiedu Formula in Reversing Gastric Mucosa Carcinogenesis in Rats with Chronic Atrophic Gastritis Based on ERK/Cyclin E Pathway

目的探讨健脾化瘀解毒方对慢性萎缩性胃炎(CAG)大鼠增殖、分化ERK/Cyclin E信号通路的影响。方法选用SPF级SD大鼠,运用N-甲基-N’-硝基-N-亚硝基胍(MNNG)复合造模法复制CAG大鼠模型,设立分组:正常组,模型组,叶酸组,健脾化瘀解毒方高、中、低组。HE染色观察大鼠胃黏膜组织病理学改变;采用qRT-PCR法检测胃黏膜组织ERK1、ERK2、Cyclin E mRNA转录水平;免疫组化法检测胃黏膜组织ERK1、ERK2、Cyclin E蛋白表达。结果健脾化瘀解毒方可显著改善CAG大鼠胃黏膜上皮腺体结构、形态紊乱及细胞异型性等病理表现。模型组CAG大鼠胃黏膜上皮的ERK1、ERK2、Cyclin E mRNA水平及蛋白表达评分均较正常组显著增加(P<0.05或P<0.01);与模型组比较,健脾化瘀解毒方能显著下调CAG大鼠胃黏膜上皮的ERK1、Cyclin E mRNA水平及蛋白表达评分(P<0.05,P<0.01),以高剂量组疗效最佳;健脾化瘀解毒方对ERK2无显著调控作用。结论健脾化瘀解毒方能显著下调ERK1、Cyclin E的表达,抑制胃黏膜上皮细胞的过度增殖,进而阻断CAG恶性转变。

Objective To investigate the effects of Jianpi Huayu Jiedu Formula on the expressions of ERK1,ERK2 and Cyclin E in chronic atrophic gastritis(CAG)rats.Methods SPF-class SD rats were used to replicate the CAG rat model by N-methyl-N’-nitro-N-nitrosoguanidine(MNNG)composite modeling.The animals were randomly allocated into six groups as follows:normal group,model group,folic acid group,high,medium and lowdose groups which were treated with different doses of Jianpi Huayu Jiedu Formula by gavage.The pathological changes of gastric mucosa were observed by HE staining.The mRNA levels of ERK1,ERK2 and Cyclin E mRNA in gastric mucosa were detected by qRT-PCR.The expressions of ERK1,ERK2 and Cyclin E in gastric mucosa were detected by immunohistochemistry.Results The mRNA levels and protein expression levels of ERK1,ERK2 and Cyclin E in gastric mucosal epithelium of model group were significantly increased compared with those of the normal group(P<0.05 or P<0.01).Compared with those of the model group,Jianpi Huayu Jiedu Formula group’s levels of ERK1 and Cyclin E mRNA and protein expression in gastric mucosal epithelium of CAG rats were significantly down-regulated(P<0.05 or P<0.01).Jianpi Huayu Jiedu Formula had no significant regulatory effect on ERK2.Conclusion Jianpi Huayu Jiedu Formula can significantly down-regulate the expressions of ERK1 and Cyclin E,inhibit the excessive proliferation of cells,and then block the malignant transformation of CAG.