摘要
目的:观察参苓白术散通过TLR4/MyD88通路对Lewis肺癌小鼠肿瘤组织凋亡的干预效果和机制,探讨培土生金法对肺癌的部分治疗机制。方法:40只SPF级雄性C57小鼠,采用腋窝皮下接种肺癌Lewis细胞悬液法建立Lewis肺癌小鼠模型,造模成功后,随机分成Lewis肺癌组、顺铂组、中药组和中药+顺铂组,每组10只。根据人的使用剂量,按照体表面积折算法进行人和小鼠等效剂量换算。各药物组小鼠给予相应药物,Lewis肺癌组小鼠给予等体积生理盐水。给药14 d后,采用TUNEL染色法检测小鼠肿瘤组织细胞凋亡情况,并计算细胞凋亡指数;采用免疫组化法测定Toll样受体4(TLR4)、MyD88和Caspase-3的蛋白表达水平;采用Western blotting法测定TLR4、MyD88和Caspase-3蛋白表达水平。结果:与Lewis肺癌组比较,顺铂组肿瘤增长抑制率为38.59%,肿瘤细胞凋亡指数升高,TLR4和MyD88阳性细胞数量减少,平均光密度值明显降低(P<0.01),蛋白表达水平降低(P<0.01),Caspase-3阳性细胞数量增加,平均光密度值升高(P<0.01),蛋白表达水平升高(P<0.01);中药组肿瘤增长抑制率为30.14%,肿瘤细胞凋亡指数升高,TLR4和MyD88阳性细胞数量减少,平均光密度值降低(P<0.01),蛋白表达水平降低(P<0.01),Caspase-3阳性细胞数量增加,平均光密度值升高(P<0.01),蛋白表达水平升高(P<0.01);中药+顺铂组肿瘤增长抑制率为44.51%,肿瘤细胞凋亡指数升高,TLR4和MyD88阳性细胞数量减少,平均光密度值降低(P<0.01),蛋白表达水平降低(P<0.01),Caspase-3阳性细胞数量增加,平均光密度值升高(P<0.01),蛋白表达水平升高(P<0.01)。与顺铂组、中药组比较,中药+顺铂组肿瘤细胞凋亡指数升高,TLR4、MyD88和Caspase-3蛋白表达水平的变化更明显。结论:参苓白术散可以降低Lewis肺癌小鼠TLR4、MyD88蛋白表达水平,增加Caspase-3蛋白表达水平,通过调控TLR4/MyD88通路,促进肿瘤细胞凋亡。
Objective: To observe the intervention effect and mechanism of Shenling Baizhu powder on tumor tissue apoptosis in Lewis lung cancer mice through TLR4/MyD88 pathway, and to explore therapeutic mechanisms of Peitu Shengjin method on lung cancer. Methods: Forty SPF male C57 mice were inoculated subcutaneously into the armpit with a suspension of lung cancer cells to establish a Lewis lung cancer mouse model.After successful modeling, they were randomly divided into 4 groups: Lewis lung cancer group, cisplatin group,traditional Chinese medicine group and traditional Chinese medicine + cisplatin group, with 10 mice in each group. According to the human dosage, the equivalent calculation of human and mouse was carried out according to the body surface area conversion algorithm. Mice in each drug group were given corresponding drugs, and mice in Lewis lung cancer group were given equal volume normal saline. After 14 days of administration,TUNEL staining was used to detect the apoptosis of tumor tissues in mice, and the apoptosis index was calculated.The protein expression levels of toll-like receptor 4(TLR4), MyD88 and Caspase-3 were measured by immunohistochemistry. The expression levels of TLR4, MyD88 and Caspase-3 were measured by Western blotting.Results: Compared with Lewis lung cancer group, in cisplatin group, the tumor growth inhibition rate was 38.59%, the tumor cell apoptosis index increased, the number of TLR4 and MyD88 positive cells and the average optical density decreased(P<0.01), the protein expression level decreased(P<0.01), the number of Caspase-3 positive cells increased(P<0.01), the average optical density increased(P<0.01), and the protein expression level increased(P <0.01). In traditional Chinese medicine group, the tumor growth inhibition rate was 30.14%, the tumor cell apoptosis index increased, the number of TLR4 and MyD88 positive cells and the average optical density decreased(P<0.01), the protein expression level decreased(P<0.01), the number of Caspase-3 positive cells increased, the average optical density increased(P<0.01), and the protein expression level increased(P<0.01). In traditional Chinese medicine + cisplatin group, the tumor growth inhibition rate was 44.51%, the tumor cell apoptosis index increased, the number of TLR4 and MyD88 positive cells and the average optical density decreased(P<0.01), the protein expression level decreased(P<0.01), the number of Caspase-3 positive cells increased, the average optical density increased(P <0.01), and the protein expression level increased(P <0.01).Compared with cisplatin group and traditional Chinese medicine group, the apoptosis index of tumor cells in traditional Chinese medicine + cisplatin group was higher, and the changes of TLR4, MyD88 and Caspase-3 protein expression levels were more obvious. Conclusion: Shenling Baizhu powder can decrease the protein expression level related TLR4 and MyD88 and increase the protein expression level related Caspase3. It can promote the apoptosis of Lewis lung cancer through TLR4/MyD88 pathway.
作者
张云亭
刘羽茜
蒋宗蓥
董秀
张林
王靖宇
王艳杰
ZHANG Yun-ting;LIU Yu-xi;JIANG Zong-ying;DONG Xiu;ZHANG Lin;WANG Jing-yu;WANG Yan-jie(College of Integrated Chinese and Western Medicine,Liaoning University of Traditional Chinese Medicine,Shenyang Liaoning 110847,China)
出处
《中医药导报》
2021年第11期41-45,63,共6页
Guiding Journal of Traditional Chinese Medicine and Pharmacy
基金
辽宁省科学技术计划项目(自然基金指导计划)(2019-ZD-0950)。
