益气活血方对胃癌前病变大鼠胃黏膜Sonic Hedgehog信号通路的影响

Effects of Yiqi Huoxue Decoction on the expression of Sonic Hedgehog signaling pathway in the stomach mucosa of rats with gastric precancerous lesions

目的:研究益气活血方对1-甲基-3-硝基-1-亚硝基胍(MNNG)诱导的新生大鼠胃癌前病变胃黏膜组织Sonic Hedgehog(Shh)信号通路的影响。方法:48只雄性乳鼠随机分为正常组8只,造模组40只。造模组灌服800mg/L的MNNG 0.1mL/只,正常组灌服等量0.9%氯化钠溶液,共10d。23周后将造模组随机均分为模型组,中药高、中、低剂量组,环耙明组。正常组、模型组灌服0.9%氯化钠溶液10mL/kg,中药高、中、低剂量组分别灌服38.4、19.2、9.6g/kg的益气活血方流浸膏,环耙明组腹腔注射0.2mg/kg环靶明,均每日1次,12周后HE染色观察胃黏膜组织病理学变化,免疫组化检测胃黏膜PCNA的表达,qRT-PCR法检测胃黏膜组织Shh、Ptch1、Smo、Gli1、SuFu、CyclinD1、CyclinE1、C-Myc mRNA的表达,Western Blot检测胃黏膜组织Shh、Ptch1、Smo、Gli1、SuFu、CyclinD1、CyclinE1、C-Myc、p-c-Myc蛋白表达。结果:与模型组比较,益气活血方高、中、低剂量组和环耙明组大鼠胃黏膜炎症及异型增生程度均显著减轻,PCNA表达水平显著降低(P<0.05,P<0.01),Ptch1及SUFU mRNA表达显著升高(P<0.05,P<0.01),Shh、Smo、Gli-1、CyclinD1及p-c-Myc蛋白的相对表达量显著降低而Ptch1及SUFU蛋白的表达显著升高(P<0.05,P<0.01)。结论:益气活血方可能通过下调Shh信号通路的表达起到对胃癌前病变的防治作用。

Objective:To investigate the impact of Yiqi Huoxue Decoction on the expression of Sonic Hedgehog(Shh)signaling pathway in the stomach mucosa of rats with gastric precancerous lesions induced by MNNG.Methods:A total of 48 suckling male rats were randomly divided into 2 groups,8 in normal group and 40 in model group.The rats in the model group were administrated with 0.1 mL MNNG at the dosage of 800 mg/L for 10 days,while the rats in the normal group with normal saline.The rats of the model group were randomly divided into model group,high,middle,low dosage group of traditional Chinese medicine(TCM),and cyclopamine group.Then the rats of normal group and model group were given saline at the dosage of 10 mL/kg,while the three TCM groups were administrated with Yiqi Huoxue prescription at the dosage of 38.4,19.2,9.6 g/kg,rats in the cyclopamine group were injected intraperitoneally with cyclopamine at the dosage of 0.2 mg/kg,respectively.All the rats were killed 12 weeks later.Histopathologic changes of the gastric mucosa were observed by HE staining;expression of PCNA in the gastric mucosa was determined by immunohistochemistry;the levels of Shh,Ptch1,Smo,Gli1,Su-Fu,CyclinD1,CyclinE1,c-Myc mRNA in the gastric mucosa were determined by qRF-PCR;the expressions of Shh,Ptch1,Smo,Gli1,SuFu,CyclinD1,c-Myc,p-c-Myc protein were detected by Western Blot.Results:Compared with the model group,rats of high,middle,low dosage group of TCM and cyclohexane group showed significantly less gastric mucosa inflammation and lower degree of dysplasia,lower expression of PCNA(P<0.05,P<0.01),significantly increased levels of Ptch1 and SUFU mRNA(P<0.05,P<0.01),and similarly,lower expression of Shh,Smo,Gli-1,CyclinD1 and p-c-Myc protein and higher expression of Ptch1 and SUFU protein in the gastric mucosa(P<0.05,P<0.01).Conclusion:Yiqi Huoxue Decoction may play a preventive role in gastric precancerous lesions by down-regulating the expression of Shh signaling pathway.