摘要
目的:观察中药癌痛方乙醇提取物对Lewis肺癌小鼠肿瘤细胞的影响,以及通过降钙素基因相关肽(Calcitonin Gene-related Peptide,CGRP)/p-环磷腺苷效应元件结合蛋白(cAMP-response Element Binding Protein,CREB)信号通路调控肺癌细胞凋亡的分子作用机制。方法:采取皮下注射肿瘤细胞的方法构建C57BL/6小鼠Lewis肺癌小鼠模型,随机分为癌痛方乙醇提取物组、模型对照组,分别给予癌痛方乙醇提取物或等量0.9%氯化钠溶液连续灌胃14天后,依次处死、称重、剥离瘤体、称量瘤重、测量肿瘤体积。HE染色方法观察肿瘤组织细胞的形态学特征,免疫荧光标记法分析细胞凋亡和增殖水平,蛋白质免疫印迹法检测肿瘤组织中相关蛋白表达及CGRP/p-CREB信号通路的影响。结果:比较各组瘤重,癌痛方乙醇提取物组小于模型对照组,但差异无统计学意义(P>0.05)。癌痛方乙醇提取物组肿瘤体积小于模型对照组,差异有统计学意义。光镜下观察肿瘤组织切片可见模型对照组肿瘤细胞分布均匀致密;同时,癌痛方乙醇提取物组肿瘤细胞肿胀,可见多核细胞。在肿瘤细胞凋亡和增殖水平上,癌痛方乙醇提取物组能显著提高小鼠肿瘤组织凋亡蛋白表达水平,促进细胞凋亡(P<0.05),抑制小鼠肿瘤组织增殖(P<0.05);蛋白质免疫印迹法检测结果提示癌痛方乙醇提取物可明显下调CGRP及p-CREB蛋白表达水平。结论:中药癌痛方乙醇提取物可诱导Lewis肺癌小鼠肿瘤细胞的凋亡,在一定程度上减缓肺癌细胞的增殖,其作用机制可能与CGRP/p-CREB信号通路的抑制有关。
Objective:To observe the effects of ethanol extract of the Aitong prescription(癌痛方)on tumor cells of Lewis lung cancer mice,and the molecular mechanism of CGRP/p-CREB signal pathway on apoptosis of lung cancer cells.Methods:C57BL/6 mice model of Lewis lung cancer was established by subcutaneous injection of tumor cells.The mice were randomly divided into the Aitongfang ethanol extract group and the model control group.After 14 days of intragastric administration with ethanol extract of the Aitong prescription or the same amount of normal saline,respectively,the tumors model mice were killed and weighed,the tumor was removed and weighed,and then the tumor volume was measured.The morphological characteristics of tumor cells were observed by HE staining method.The level of cell apoptosis and proliferation was analyzed by immunofluorescence method.The expression of related proteins and the effects of CGRP/p-CREB signal pathway were detected by Western blot method.Results:The tumor weight in the Aitongfang ethanol extract group was smaller than the model control group,but there was no significant difference between the groups(P>0.05).The tumor volume in the Aitongfang ethanol extract group was smaller than the model control group,and there was a statistically significant difference.The tumor tissue sections under light microscope showed that the tumor cells in the model control group were enenly and densely distributed,the tumor cells in the Aitongfang ethanol extract group were swollen,and multinucleated cells were seen.In the Aitongfang ethanol extract group,the expression level of apoptotic protein in mice tumor tissue was obviously increased,cell apoptosis was promoted(P<0.05),and the proliferation of mice tumor tissues was inhibited(P<0.05).The results of Western blotting indicated that ethanol extract of the Aitong prescription could significantly decrease the expression of CGRP and p-CREB protein.Conclusion:The ethanol extract of the Aitong prescription can induce the apoptosis of Lewis lung cancer mice c
出处
《中医临床研究》
2021年第28期6-10,共5页
Clinical Journal Of Chinese Medicine
关键词
肺癌细胞
癌痛方
LEWIS肺癌小鼠
信号通路
Lung cancer cell
The Aitong prescription
Lewis lung cancer mice
Signal pathway
