摘要
目的:基于肝细胞生长因子及其受体(HGF/C-Met)信号通路探讨乌梅丸对寒热错杂证Lewis肺癌小鼠的抑癌效果及作用机制。方法:将20只健康雄性小鼠分为正常组、模型组、顺铂组和乌梅丸组,每组5只。通过丙硫氧嘧啶+知母石膏水煎液+番泻叶灌胃、冰水游泳与干酵母混悬液皮下注射,联合Lewis细胞悬液右侧腋窝皮下注射的方式制备寒热错杂证Lewis肺癌小鼠模型。模型制备完成后,顺铂组予以4.0 mg·kg^(-1)顺铂腹腔注射,乌梅丸组予以12.5 mL·kg^(-1)乌梅丸灌胃,模型组予以等体积蒸馏水灌胃,连续6周。取材检验,测量和计算瘤质量、瘤体积、抑癌率和抑制肺癌转移率,采用苏木素-伊红(HE)染色观察肺癌组织病理学形态,采用免疫组化法分析肿瘤组织生长状态,采用实时荧光定量聚合酶链式反应(Real-time PCR)检测HGF、C-Met mRNA表达,采用蛋白免疫印迹法(Western blot)检测HGF、C-Met、存活素(Survivin)和X连锁凋亡抑制蛋白(XIAP)的蛋白表达。结果:与正常组比较,模型组HGF、C-Met mRNA表达显著升高(P<0.01),HGF、C-Met、Survivin和XIAP的蛋白表达显著升高(P<0.01)。与模型组比较,乌梅丸组的阳性细胞比率、阳性细胞密度、阳性评分降低(P<0.05),组织化学评分、瘤质量、瘤体积显著降低(P<0.01),HGF、C-Met mRNA表达显著降低(P<0.01),HGF、C-Met、Survivin和XIAP的蛋白表达显著降低(P<0.01)。与模型组比较,顺铂组的阳性细胞率、阳性细胞密度明显降低(P<0.05),阳性评分、瘤质量、瘤体积显著降低(P<0.01),HGF、C-Met mRNA表达显著降低(P<0.01),HGF、C-Met、Survivin、XIAP蛋白表达显著降低(P<0.01);组织化学评分相当,差异无统计学意义。与顺铂组比较,乌梅丸组HGF mRNA表达升高(P<0.01);阳性细胞率、阳性细胞密度、组织化学评分、阳性评分、瘤质量、瘤体积相当,C-Met mRNA表达及HGF、C-Met、Survivin、XIAP蛋白表达相当,差异均无统计学意义。结论:乌梅丸能够通过抑制HGF/C-Met信号通路减缓寒热错杂证Lewis肺癌小鼠的肺癌进展。
Objective:To investigate the effect and mechanism of Wumeiwan against Lewis lung cancer in mice with syndrome of cold and heat in complexity based on hepatocyte growth factor/mesenchymalepithelial transition factor(HGF/C-Met)signaling pathway.Method:Twenty healthy male mice were classified into blank group,model group(equivalent volume of distilled water,ig),cisplatin group(4.0 mg·kg^(-1)cisplatin,ip),and Wumeiwan group(12.5 mL·kg^(-1)Wumeiwan,ig),with 5 in each group.Lewis lung cancer with the syndrome of cold and heat in complexity was induced in mice except the blank group by gavage of propylthiouracil,Zhimu Shigaotang,and Fanxieye,ice-water swimming,and subcutaneous injection of dry yeast suspension and Lewis cell suspension under the right armpit.After modeling,administration began and lasted 6 weeks.After the experiment,the tumor weight,tumor volume,tumor inhibition rate,and lung cancer metastasis-inhibiting proportion were measured and calculated.The pathological morphology of lung tissue was observed based on hematoxylin and eosin(HE)staining.The growth state of tumor tissue was analyzed by immunohistochemistry.The mRNA expression of HGF and C-Met was detected by Real-time polymerase chain reaction(PCR),and the protein expressions of HGF,C-Met,survivin,and X-linked inhibitor of apoptosis protein(XIAP)by Western blot.Result:Compared with the blank group,the model group showed high mRNA expression of HGF and C-Met and protein expression of HGF,C-Met,surviving,and XIAP(P<0.01).Compared with the model group,Wumeiwan group displayed low proportion of positive cells,positive cell density,positive score(P<0.05),histochemical score,tumor weight,tumor volume(P<0.01),mRNA expression of HGF and C-Met(P<0.01),and protein expression of HGF,C-Met,surviving,and XIAP(P<0.01).Compared with the model group,the cisplatin group displayed decrease in the proportion of positive cells,density of positive cells(P<0.05),positive score,tumor weight,tumor volume(P<0.01),mRNA expression of HGF and C-Met(P<0.01),and protein expression of HGF,C-Met,surviving,and XIAP(P<0.01),and insignificant variation in the histochemical score.Wumeiwan group had high mRNA expression of HGF(P<0.01),and insignificant variation in the proportion of positive cells,positive cell density,histochemical score,positive score,tumor weight,tumor volume,mRNA expression of C-Met,and protein expression of HGF,C-Met,surviving,and XIAP.Conclusion:Wumeiwan can slow down the progression of Lewis lung cancer in mice with syndrome of cold and heat I complexicity by inhibiting HGF/C-Met signaling pathway.
作者
俞赟丰
上官雪丽
李新
YU Yunfeng;SHANGGUAN Xueli;LI Xin(Hunan University of Chinese Medicine,Changsha 410208,China;The Second Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410005,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2022年第21期32-41,共10页
Chinese Journal of Experimental Traditional Medical Formulae
基金
湖南省教育厅科学研究项目(16A161)。
关键词
乌梅丸
寒热错杂
肺癌
肝细胞生长因子及其受体(HGF/C-Met)
Wumeiwan
cold and heat in complexity
lung cancer
hepatocyte growth factor/mesenchymal-epithelial transition factor(HGF/C-Met)
作者简介
第一作者:俞赟丰,硕士,医师,从事中医药防治脾胃病研究,E-mail:1091047331@qq.com;通信作者:李新,硕士,副教授,从事中医药防治呼吸系统疾病研究,E-mail:lixin2007@163.com。
