仁术健胃颗粒含药血清对MC细胞增殖及凋亡相关因子表达的影响  

Effect of serum-containing Renshu Jianwei granule on the expression of proliferation and apoptosis related factors in MC cells

作　　者：张廷 孙惠丽 严士海[1] 朱长乐[1] 陆为民[1] ZHANG Ting;SUN Huili;YAN Shihai;ZHU Changle;LU Weimin(Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,Jiangsu,China;NanTong Hospital of Traditional Chinese Medicine,Nantong 226001,Jiangsu,China)

机构地区：[1]南京中医药大学附属医院,江苏南京210029 [2]江苏省南通市中医院,江苏南通226001

出　　处：《现代中西医结合杂志》2020年第34期3776-3781,3822,共7页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：国家中医药管理局“国医大师徐景藩学术经验传承研究室”建设项目(国中医药人教发〔2010〕59号);江苏省卫生健康委干部保健课题(BJ18025)。

摘　　要：目的观察仁术健胃颗粒含药血清对MC细胞增殖及凋亡相关因子表达的影响,从分子生物学角度探讨该方治疗胃癌前病变的机制。方法将15只SPF级新西兰大白兔随机分为5组,每组3只。生理盐水组灌胃生理盐水,维A酸组灌胃0.5 mg/mL的维A酸,仁术健胃颗粒低、中、高剂量组分别灌胃0.25 g/mL、0.5 g/mL、1 g/mL的仁术健胃颗粒,均连续灌胃3 d。末次灌胃3 h后,于耳缘静脉取血,获得含药血清。用含2×10^-5 mol/L MNNG的完全培养液培养GES-1细胞24 h后换用正常完全培养液,一段时间后获得MC细胞模型。细胞实验分为6组,GES-1细胞组、MC细胞组加入含生理盐水组含药血清培养,维A酸组及仁术健胃颗粒低、中、高剂量组分别加入相应组含药血清培养,运用MTT法检测各组MC细胞增殖情况,SP法检测各组MC细胞中CDK4、P16、Fas、Bcl-2、P53蛋白表达情况。结果5%含药血清维A酸组及仁术健胃颗粒中剂量组、10%及20%含药血清维A酸组及仁术健胃颗粒各剂量组MC细胞增殖OD值均明显低于MC细胞组(P均<0.05),10%含药血清仁术健胃颗粒高剂量组MC细胞增殖OD值明显低于维A酸组(P<0.05)。与MC细胞组比较,维A酸组和仁术健胃颗粒各剂量组中CDK4蛋白阳性表达率均明显降低(P均<0.05),P16、Fas蛋白阳性表达率均明显升高(P均<0.05);且维A酸组及仁术健胃颗粒高剂量组Bcl-2、P53蛋白阳性表达率和仁术健胃颗粒中剂量组Bcl-2蛋白阳性表达率也均明显低于MC细胞组(P均<0.05)。结论仁术健胃颗粒能抑制MC细胞的增殖,可抑制CDK4、Bcl-2、P53蛋白表达及促进P16、Fas蛋白表达,发挥抗增殖及促凋亡的作用,这可能是其阻断或逆转胃癌前病变的作用机制之一。Objective It is to observe the effect of serum containing Renzhu Jianwei granules on the expression of proliferation and apoptosis-related factors in MC cells,and to explore the mechanism of this decoction in the treatment of gastric precancerous lesions from the perspective of molecular biology.Methods 15 SPF New Zealand white rabbits were randomly divided into 5 groups,3 rabbits in each group.The normal saline group was intragastrically administered with normal saline,the tretinoin group was intragastrically administered with 0.5 mg/mL tretinoin,and the Renzhu Jianwei granule low,medium,and high dose groups were respectively intragastrically administered with 0.25 g/mL,0.5 g/mL,1 g/mL Renzhu Jianwei granules,all the groups were administered continuously for 3 days.At 3 hours after the last gavage,the blood was taken from the ear vein to obtain medicated serum.GES-1 cells were cultured with a complete culture medium containing 2×10^-5 mol/L MNNG for 24 hours which were replaced with a normal complete medium.After a period of time,the MC cell models were obtained.The cell experiment was divided into 6 groups.The GES-1 cell group and MC cell group were cultured with the medicated serum gotten from the normal saline group,and the tretinoin group and the low,medium and high dose groups of Renzhu Jianwei granules were cultured with the respective medicated serum gotten from each group.The proliferation of MC cells in each group was detected by MTT method,the expression of CDK4,P16,Fas,Bcl-2,P53 protein of MC cells in each group was detected by SP method.Results The OD value of MC cell proliferation in 5%medicated serum retinoic acid group and medium dose Renzhu Jianwei granule group,10%and 20%medicated serum retinoic acid group and each dose Renzhu Jianwei granule groups were significantly lower than that in MC cell group(all P<0.05),the OD value of MC cell proliferation in the 10%medicated serum high-dose Renzhu Jianwei granule group was significantly lower than that in the tretinoin group(P<0.05).Compared with the

关 键 词：仁术健胃颗粒 MC细胞 CDK4 P16 FAS BCL-2 P53 

分 类 号：R-332[医药卫生]