基于Sonic Hedgehog通路探讨复方胃炎合剂干预慢性萎缩性胃炎癌前病变的机制

Based on Sonic Hedgehog Pathway to Explore the Mechanism of Compound Gastritis Mixture on Precancerous Lesions of Gastric Cancer

目的观察复方胃炎合剂对慢性萎缩性胃炎癌前病变(precancerous lesions of gastric cancer,PLGC)脾虚痰湿热瘀证大鼠Sonic Hedgehog(Shh)通路相关基因表达的影响,并探讨其抑制PLGC进展的机制。方法SPF级雄性Wistar大鼠随机分为空白组、空白+中药组、模型组、维酶素组、中药低剂量组、中药中剂量组和中药高剂量组,每组8只。造模组采用复合造模法构建PLGC脾虚痰湿热瘀证大鼠模型,造模成功后,各组予以相应干预,干预30 d后进行标本采集,采用HE染色检测大鼠胃黏膜病变情况,RT-qPCR检测胃组织Shh通路相关基因SHH、PTCH1、SMO、Gli1、Gli2、Gli3 mRNA的表达。结果HE染色提示空白组和空白+中药组大鼠胃组织病理组织学正常,模型组大鼠胃黏膜萎缩,并可见肠上皮化生(intestinal metaplasia,IM)和上皮内瘤变(intraepithelial neoplasia,IN);维酶素组和中药各剂量组病理组织学可见不同程度改善,以中药中、高剂量组改善最为明显。与空白组比较,模型组大鼠SHH、SMO、PTCH、Gli1 mRNA表达量下降,Gli2、Gli3 mRNA表达量上升,差异均有统计学意义(P<0.05,P<0.01)。与模型组比较,各治疗组大鼠胃组织SHH、SMO、PTCH、Gli1 mRNA表达量均上升,Gli2、Gli3 mRNA表达量均降低,其中空白+中药组、维酶素组、中药中剂量组、中药高剂量组SHH、SMO、PTCH mRNA表达差异有统计学意义(P<0.05,P<0.01),中药高剂量组大鼠胃组织Gli1 mRNA表达差异有统计学意义(P<0.05),空白+中药组、维酶素组、中药中剂量组、中药高剂量组大鼠胃组织Gli2 mRNA表达差异有统计学意义(P<0.01),中药各剂量组Gli3 mRNA表达差异无统计学意义(P>0.05)。结论复方胃炎合剂可通过激活Shh通路改善胃黏膜病变,从而抑制PLGC的进展。

Objective To observe the effect of compound gastritis mixture on the related genes expression of Sonic Hedgehog(Shh)pathway in rats with spleen deficiency,dampness-heat and blood stasis syndrome of precancerous lesions of gastric cancer(PLGC),and to explore its mechanism of inhibiting the progression of PLGC.Methods SPF male Wistar rats were randomly divided into the blank group,the blank+traditional Chinese medicine(TCM)group,the model group,the vitacoenzyme group,the low-dosage TCM group,the medium-dosage TCM group,and the high-dosage TCM group,with 8 rats in each group.The rat model of spleen deficiency,dampness-heat,and blood stasis syndrome of PLGC was established by compound modeling method.After the model was successfully established,each group was given 30 days of related interventions.Then,stomach specimens were collected and the pathological changes of gastric mucosa were detected by HE staining,and the related genes expression of Shh pathway of SHH,PTCH,SMO,Gli1,Gli2 and Gli3 mRNA in gastric tissue was detected by RT-qPCR.Results HE staining showed that the histopathology of gastric tissue in the blank group and the blank+TCM group was normal,however,atrophy,intestinal metaplasia(IM)and intraepithelial neoplasia(IN)could be seen in the gastric mucosa of rats in the model group;the histopathology of the vitacoenzyme group and each dose of TCM group were improved in varying degrees,especially in the medium-dosage TCM group and the high-dosage TCM group.Compared with the blank group,the expressions of SHH,SMO,PTCH and Gli1 mRNA in the model group decreased,while the expressions of Gli2 and Gli3 mRNA increased,and the differences were statistically significant(P<0.05,P<0.01).Compared with the model group,the expressions of SHH,SMO,PTCH and Gli1 mRNA in gastric tissue of rats in each treatment group increased,while the expressions of Gli2 and Gli3 mRNA decreased,among them,there were significant differences in the expressions of SHH,SMO and PTCH mRNA in the blank+TCM group,the vitacoenzyme group,the medium-dosage TCM group,and the high-dosage TCM group(P<0.05,P<0.01),there were significant difference in the expression of Gli1 mRNA in gastric tissue of rats in the high-dosage TCM group(P<0.05),the expression of Gli2 mRNA in gastric tissue of rats in the blank+TCM group,the vitacoenzyme group,the medium-dosage TCM group,and the high-dosage TCM group was decreased significantly(P<0.01),while the expression of Gli3 mRNA in each dosage TCM group was not statistically significant(P>0.05).Conclusion Compound gastritis mixture can improve the pathological change of gastric mucosa by reactivating Shh pathway,thus inhibiting the progression of PLGC.