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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 Dihuang Guan Shi Tong Granule on the apoptosis of esophageal cancer-induced rat and the expression of proto-oncogene (c-myc) and telomerase reverse transcriptase (TERT) after esophageal cancer, and to analyze its molecular biology Mechanism. Methods: Clean Wistar rats were selected, except the blank control group (normal group), the other rats were methyl amyl nitrosamines 5 mg.kg-1 sc, once a week for 18 weeks, After the model was successfully established, rats in the model group were anesthetized with pentobarbital sodium and local irradiation with cobalt 60 radiotherapy machine. After 24 h of the last irradiation, the rats were randomly divided into radiotherapy group, Rehmannia glutinosa tube fed high, medium and low dose group, and Liuweidihuangwan (positive control 4.5g.kg-1) group, respectively, for 35 consecutive days. TUNEL method was used to detect apoptosis of esophageal cancer cells. Immunohistochemistry was used to detect the expression of c-myc and TERT protein in esophageal cells. Results: ①TUNEL assay showed that all the groups showed apoptosis, and the apoptotic index in each group increased obviously, while the apoptosis in high dose group increased obviously. ② The results of immunohistochemistry showed that the expression of c-myc protein in model group and radiotherapy group were significantly higher than those in the control group (P <0.05). The high dose Group and radiotherapy group was statistically significant (P <0.05). ③ Compared with the radiotherapy group, the expression of TERT protein in each drug group showed a decreasing trend, and there was a significant difference between the high, middle and low dose groups (P <0.01, P <0.05). CONCLUSION: Induction of apoptosis is one of the mechanisms of Dihuang Guan Shi Tong granule in killing esophageal cancer cells in vivo, and this mechanism is related to the inhibition of the expression of c-myc and TERT proteins. The drug can also effectively improve the esophageal carcinoma after chemotherapy Side effects, and prevent recurrence.