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目的:观察益气活血解毒中药胃痞消对胃癌前病变(PLGC)大鼠Notch/DLLs信号通路及其下游靶蛋白的调控效应和机制。方法:雄性SPF级SD大鼠随机分为空白组,模型组,维酶素组(0.2g·kg-1·d-1)、胃痞消高、中、低剂量(15、7.5、3.75g·kg-1·d-1)组6组,复制脾虚证PLGC大鼠模型,连续给药10周,免疫组化法检测大鼠胃黏膜上皮细胞(GECs)周期素E(cyclin E)、Myc转录蛋白(c-myc)和氨酸重复序列的G蛋白偶联受体5(Lgr5)表达,RT-PCR法测GECs Notch1、Notch4和Delta样配体4(DLL4)mRNA的表达。结果:与空白组比较,PLGC大鼠GECs Notch1 mRNA和cyclin E、c-myc与Lgr5蛋白表达均显著性增加(P<0.01),Notch4 mRNA显著性减小(P<0.01),PLGC大鼠Lgr5胃癌干细胞由基底侧向胃腔侧侵袭迁移;胃痞消3剂量组GECs Notch1、Notch4mRNA及cyclin E、c-myc和Lgr5蛋白表达累积面积百分比和平均灰度均显著性减小(P<0.01),胃痞消中、低剂量组GECs DLL4 mRNA均显著性减少(P<0.01);胃痞消低剂量组Notch1、Notch4 mRNA及cyclin E、Lgr5蛋白表达累积面积百分比和平均光度较胃痞消高、中剂量组均显著性减小(P<0.01,P<0.05)。结论:胃痞消可拮抗Notch/DLLs通路活化诱导cyclin E、c-myc和Lgr5上调所致PLGC大鼠胃癌干细胞的增殖、凋亡抑制、侵袭和迁移。
Objective: To observe the regulatory effect and mechanism of Yiwei Huoxue Huoxue Decoction Chinese medicine Weibixiao on Notch / DLLs signal pathway and its downstream target protein in gastric precancerous lesions (PLGC) rats. Methods: Male SPF SD rats were randomly divided into blank group, model group, verapamil group (0.2g · kg-1 · d-1), Weipi disappeared high, medium and low dose (15,7.5,3.75g · Kg-1 · d-1) for 6 weeks. The rats were injected with PLGC for 10 weeks. The expressions of cyclin E, Myc The expression of Notch1, Notch4 and Delta-like ligand 4 (DLL4) mRNA in GECs was detected by RT-PCR. Results: Compared with the blank group, the expressions of Notch1 mRNA, cyclin E, c-myc and Lgr5 protein in GGCs were significantly increased (P <0.01), Notch4 mRNA was significantly decreased in PLGC rats (P <0.01) The gastric cancer stem cells migrated from the basolateral side to the gastric cavity side. The accumulative area percent and the average gray level of Notch1, Notch4 mRNA, cyclin E, c-myc and Lgr5 in Giemsa group were significantly decreased (P <0.01) (P <0.01). The accumulative area percentage and average luminosity of Notch1, Notch4 mRNA and cyclin E, Lgr5 protein expression in Weipixiao low dose group were significantly lower than those in Weipi group , Medium dose group were significantly reduced (P <0.01, P <0.05). Conclusion: Weipixiao can antagonize the activation of Notch / DLLs pathway and induce the proliferation, apoptosis inhibition, invasion and migration of gastric cancer stem cells in PLGC rats induced by the upregulation of cyclin E, c-myc and Lgr5.