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目的:观察清热化湿方对人胃腺癌细胞7901增殖、凋亡及CD14基因、肿瘤坏死因子α(TNF-α)及白细胞介素1β(IL-1β)表达的影响。方法:采用血清药理学方法,对照组予胎牛血清,治疗组分别予不同体积分数(5%,10%,15%,20%,30%)清热化湿方含药血清干预体外培养的胃腺癌细胞SGC-7901,应用四甲基偶氮唑盐微量酶反应比色法(MTT法)检测细胞增殖,流式细胞仪分析细胞凋亡率、细胞周期,免疫组化、Real-time PCR技术检测CD14,TNF-α,IL-1β蛋白及mRNA表达情况。结果:与对照组比较,治疗组SGC-7901细胞存活率明显下降(P<0.05);与对照组比较,治疗组S期细胞减少、G0/G1期细胞增多,且与药物质量浓度有剂量依赖性(P<0.05)。治疗组可明显下调CD14,TNF-α,IL-1β蛋白及mRNA表达并促进细胞凋亡(P<0.05),以中剂量更为明显。结论:清热化湿方可能通过调控CD14,TNF-α,IL-1β表达,介导SGC-7901细胞S期,诱导细胞凋亡,发挥防治胃癌的效应。
Objective: To observe the effects of Qingre Huashi prescription on the proliferation and apoptosis of human gastric adenocarcinoma cell line 7901 and the expression of CD14, TNF-α and IL-1β. Methods: Serum pharmacology method was used. The control group was fed with fetal bovine serum. The treatment group was given different concentrations of 5%, 10%, 15%, 20% and 30% The proliferation of SGC-7901 cells was detected by MTT method. The apoptosis rate, cell cycle, immunohistochemistry and Real-time PCR were analyzed by flow cytometry The expressions of CD14, TNF-α and IL-1β protein and mRNA were detected. Results: Compared with the control group, the survival rate of SGC-7901 cells in the treatment group was significantly decreased (P <0.05). Compared with the control group, the number of cells in the S phase and the G0 / G1 phase cells in the treatment group were increased, (P <0.05). The treatment group could obviously downregulate the protein and mRNA expression of CD14, TNF-α, IL-1β and promote cell apoptosis (P <0.05). Conclusion: Qingre Huashi decoction can induce S phase and induce apoptosis of SGC-7901 cells by regulating the expression of CD14, TNF-α and IL-1β and exert the effect of preventing and treating gastric cancer.