摘要
目的:观察地黄管食通颗粒对食管癌造模大鼠细胞凋亡及放疗后原癌基因(c-myc),端粒酶逆转录酶(TERT)蛋白表达的影响,从分子生物学角度分析其作用机制。方法:选用清洁级Wistar大鼠,除空白对照组(正常组)外,其余大鼠均以甲基戊基亚硝胺5 mg.kg-1 sc,每周1次,连续18周,确定造模成功后,除模型组大鼠外,余造模大鼠均以戊巴比妥钠麻醉,钴60放射治疗机进行食管局部照射。末次照射24 h后,随机分为放疗组、地黄管食通高、中、低剂量组、六味地黄丸(阳性对照4.5g.kg-1)组,分别ig,连续35 d。应用TUNEL法检测对食管癌癌细胞凋亡的促进作用;免疫组化检测大鼠食管细胞中c-myc,TERT蛋白的表达。结果:①TUNEL法检测各组均见有细胞凋亡,且药物治疗各组细胞凋亡指数明显增高,高剂量组细胞凋亡明显增多。②免疫组化结果显示c-myc蛋白在模型组、放疗组表达率及表达强度均明显增高,药物治疗组均有不同程度的降低,以高剂量组降低最明显(P<0.05),高剂量组与放疗组比较有统计学意义(P<0.05)。③与放疗组比较,各药物组TERT蛋白表达均呈递减趋势,高、中、低剂量组有统计学意义(P<0.01,P<0.05)。结论:诱导细胞凋亡是地黄管食通颗粒在体内杀伤食管癌细胞的作用机制之一,而这一作用机制与抑制c-myc,TERT蛋白的表达相关;该药物还可有效改善食管癌化疗后副作用,并预防复发。
Objective：To observe the effect of a combination of traditional Chinese herbs Dihuang Guanshitong Granules on the rat model of esophageal cancer after rradiotherapy for inducing apoptosis and the c-myc,TERT protein expression.Method：Wistar rats were randomly divided into 2 groups.They were treated with MANA 5 mg·kg-1 by subsutaneous injection except control group（the normal）.After the model was successful,the model rats were treated with Intraperitoneal injection of sodium pentobarbital anesthesia and local esophageal radiation by cobalt60 except the model group rats.The rats were randomly divided into the radiotherapy group,the Dihuang Guanshitong high,middle,low dose（5,10,15 g·kg-1） groups,the Liuwei Dihuang Wan（4.5 g·kg-1） group after the last radiation.Then they were administered.TUNEL assay was used to examine the promotion on apoptosis of esophageal cancer cells.Immunohisto-chemistry（IHC） technique was used to measure the expression of c-myc and TERT protein in cells.Result：① The esophageal cancer cells were apoptic according to the result of TUNEL assay in all the groups.The granules was more obvious and the high-dosage group was the most.②Immunohistochemical results showed that c-myc protein expression in model group were increased than that in the radiation group.The drug therapy group were reduced by different degree and the high-dosage group were more obvious（P0.05）.The comparison between the high-dosage groups and radiation therapy group was significant（P0.05）.③TERT protein expression of each drug groups were tending to decrease compared with that in radiotherapy group.The high,middle,low dosage groups were significant（P0.01,P0.05）.Conclusion：Apoptosis is likely one of the mechanisms of Dihuang Guanshitong Granules to kill esophageal cancer cells.This is related with inhibiting c-myc,TERT protein expression.The granules can decrease side effects after chemotherapy and prevent of recurrence.
出处
《中国实验方剂学杂志》
CAS
北大核心
2011年第18期240-243,共4页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家中医药管理局科技公关项目(02-03JP25)
河南省科学技术厅项目(9412011Y0762)
