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目的:通过分析大鼠胃组织细胞凋亡和血浆差异代谢产物,探讨二冬膏延缓胃癌发生的作用机制。方法:大鼠随机分为正常组、模型组、维甲酸(12×10~(-3)g·kg~(-1))组、二冬膏低、高剂量(6,12 g·kg~(-1))组,除正常组外,其余大鼠ig N-甲基-N’-硝基-N-亚硝基胍(MNNG)0.2 g·kg~(-1)建立模型,10 d/次,共9次。各组每天ig相应药物,正常组ig等体积生理盐水,治疗3个月后处理动物,苏木素伊红(HE)染色分析胃癌程度及病理变化的改善;实时荧光定量PCR(Real-time PCR)测定胃组织中凋亡抑制基因B淋巴细胞瘤-2(Bcl-2)mRNA表达;脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TUNEL)检测胃组织细胞凋亡;血浆代谢组学寻找差异代谢产物。结果:与模型组比较,二冬膏低、高剂量组病变主要在良性的乳头状瘤和乳头状增生,说明二冬膏能够改善胃癌病变程度;低剂量组能明显降低胃组织Bcl-2 mRNA的表达(P<0.01),TUNEL显示二冬膏低、高剂量组胃组织细胞凋亡数比模型组高。二冬膏低剂量血浆代谢产物中发现的鞘氨醇-1-磷酸(Sphingosine-1-phosphate),20-羟基-二十烷四烯酸(20-HETE),甘氨胆酸(Glycocholic acid)等与细胞凋亡相关。结论:二冬膏能够促进胃癌变大鼠胃组织发生细胞凋亡,延缓胃癌发生。
OBJECTIVE: To investigate the mechanism of Erzhi Paste in delaying gastric carcinogenesis by analyzing apoptosis of rat gastric tissue and differential metabolites in plasma. Methods: The rats were randomly divided into normal group, model group, tretinoin group (12 × 10 -3 g · kg -1) (-1) group, except for the normal group, the other rats were given 0.2g · kg -1 N-methyl-N’-nitro-N-nitrosoguanidine (MNNG) / Times, a total of 9 times. Each group ig the corresponding drug, normal group ig equal volume of saline, after 3 months of treatment of animals, hematoxylin eosin (HE) staining analysis of gastric cancer and pathological changes; Real-time quantitative PCR (Real-time PCR) The expression of Bcl-2 mRNA in gastric tissues was detected by flow cytometry. Apoptosis of gastric tissue was detected by DNA nick end labeling (TUNEL) assay. Plasma metabolomics Find differences in metabolites. Results: Compared with the model group, the lesions of low and high dose Dongsituao were mainly in benign papillomatosis and papillary hyperplasia, which indicated that Shuangdong ointment could improve the degree of gastric cancer. The low dose group could significantly decrease the expression of Bcl-2 mRNA (P <0.01). The results of TUNEL showed that the apoptosis rate of gastric tissue in low and high dose Dong Er Gao group was higher than that in model group. Sphingosine-1-phosphate, 20-HETE, Glycocholic acid found in low-dose plasma metabolites of two-winter creams, Related to apoptosis. Conclusion: Shuangdong ointment can promote gastric cancer cell apoptosis in gastric cancer rats and delay the occurrence of gastric cancer.