摘要
目的探讨土贝母对H22荷瘤小鼠体内肿瘤的抑制作用及其机制。方法将30只雄性昆明小鼠随机分为空白对照组、模型组、低浓度组、中浓度组和高浓度组,每组6只。模型组、低浓度组、中浓度组和高浓度组小鼠皮下注射H22瘤细胞悬液0.1 mL建立肝癌模型,空白对照组小鼠皮下注射0.1 mL生理盐水。造模后第2天,高浓度组、中浓度组和低浓度组小鼠分别灌胃给予2.0、1.0、0.5 g·mL-1的土贝母水提液(0.2 mL·10 g-1),空白对照组和模型组小鼠给予等体积的蒸馏水;每日1次,共给药30 d。观察土贝母对H22荷瘤小鼠体质量、饮食量及肿瘤体积的影响;苏木精-伊红染色法观察小鼠肿瘤细胞形态学变化;免疫组织化学法检测小鼠肿瘤组织中ki67的表达情况;Western blot法检测小鼠肿瘤组织中caspase-1、caspase-3、caspase-9蛋白的表达。结果模型组及低浓度组、中浓度组、高浓度组小鼠每日进食量均少于空白对照组(P<0.05)。低浓度组、中浓度组、高浓度组小鼠每日进食量均多于模型组(P<0.05)。低浓度组小鼠每日进食量多于高浓度组和中浓度组(P<0.05);中浓度组与高浓度组小鼠每日进食量比较差异无统计学意义(P>0.05)。模型组、低浓度组、高浓度组与空白组小鼠体质量比较差异无统计学意义(P>0.05);中浓度组小鼠体质量低于空白组(P<0.05)。低浓度组、中浓度组、高浓度组小鼠体质量均显著低于模型组(P<0.05)。低浓度组、中浓度组、高浓度组小鼠体质量比较差异无统计学意义(P>0.05)。低浓度组、中浓度组、高浓度组小鼠肿瘤体积均低于模型组(P<0.05);低浓度组、中浓度组、高浓度组小鼠肿瘤体积比较差异无统计学意义(P>0.05)。模型组小鼠肿瘤组织细胞排列紧密,细胞结构完整,未见明显的细胞凋亡、坏死、肿胀。低浓度组、中浓度组、高浓度组小鼠均出现肿瘤组织细胞肿胀,细胞数量减少,间隙增宽,出现不同程度的坏死及凋亡。低浓度组、中浓度组、高浓度组小鼠肿瘤组织中Ki67阳性表达率均低于模型组(P<0.05);低浓度组小鼠肿瘤组织中ki67阳性表达率低于中浓度组(P<0.05);高浓度组与低浓度组、中浓度组小鼠肿瘤组织中Ki67阳性表达率比较差异无统计学意义(P>0.05)。低浓度组、中浓度组、高浓度组小鼠肿瘤组织中caspase-1、caspase-3、caspase-9蛋白相对表达量均高于模型组(P<0.05)。中浓度组、高浓度组小鼠肿瘤组织中caspase-1、caspase-3蛋白相对表达量低于低浓度组(P<0.05);高浓度组小鼠肿瘤组织中caspase-9蛋白相对表达量低于低浓度组(P<0.05);中浓度组小鼠肿瘤组织中caspase-9蛋白相对表达量与低浓度组比较差异无统计学意义(P>0.05)。高浓度组小鼠肿瘤组织中caspase-3蛋白相对表达量低于中浓度组(P<0.05);高浓度组与中浓度组小鼠肿瘤组织中caspase-1、caspase-3蛋白相对表达量比较差异无统计学意义(P>0.05)。结论土贝母能够增加小鼠饮食量,抑制肿瘤生长,降低肿瘤标志物Ki67的表达;其可能是通过上调caspase-1、caspase-3、caspase-9蛋白的表达来抑制肿瘤生长。
Objective To explore the inhibitory effect of Tubeimu on the tumor of H22 tumor bearing mice and its mechanism.Methods Thirty male Kunming mice were randomly divided into blank control group,model group,low concentration group,medium concentration group and high concentration group,with 6 mice in each group.The mice in the model group,low concentration group,medium concentration group and high concentration group were subcutaneously injected with 0.1 mL H22 tumor cell suspension to establish liver cancer model,the mice in the blank control group were subcutaneously injected with 0.1 mL normal saline.On the second day after modeling,the mice in the high concentration group,medium concentration group and low concentration group were given 2.0,1.0,0.5 g·mL-1 water extract of Tubeimu(0.2 mL·10 g-1)by gavage,respectively,and the mice in the blank control group and model group were given the same volume of distilled water;once a day for 30 days.The effect of Tubeimu on the body weight,daily food intake and tumor volume of H22 tumor bearing mice were observed;the morphological changes of tumor cells were observed by hematoxylin-eosin staining;the expression of Ki67 in tumor tissues of mice was detected by immunohistochemistry;the expressions of caspase-1,caspase-3 and caspase-9 protein in tumor tissues of mice were detected by Western blot.Results The average daily food intake of mice in the model group,low concentration group,medium concentration group and high concentration group was less than that in the blank control group(P<0.05).The average daily food intake of mice in the low concentration group,medium concentration group and high concentration group was more than that in the model group(P<0.05).The average daily food intake of mice in the low concentration group was more than that in the high concentration group and medium concentration group(P<0.05);there was no significant difference in the average daily food intake of mice between the medium concentration group and high concentration group(P>0.05).There was no significant difference in the body weight between the model group,low concentration group,high concentration group and the blank control group(P>0.05);the body weight of mice in the medium concentration group was lower than that in the blank control group(P<0.05).The body weights of mice in the low concentration group,medium concentration group and high concentration group were significantly lower than those in the model group(P<0.05).There was no significant difference in body weight of mice among the low concentration group,medium concentration group and high concentration group(P>0.05).The tumor volumes of mice in the low concentration group,medium concentration group and high concentration group were significantly lower than those in the model group(P<0.05),and there was no significant difference in the tumor volume of mice among the low concentration group,medium concentration group and high concentration group(P>0.05).In the model group,the tumor cells were arranged closely,the cell structure was complete,and there was no obvious apoptosis,cell necrosis and cell swelling.The tumor cells in the low concentration group,medium concentration group and high concentration group were swelling,and the number of cells decreased,the cell space widened;the tumor cells showed different degrees of necrosis and apoptosis.The positive expression rates of Ki67 in tumor tissue of mice in the low concentration group,medium concentration group and high concentration group were lower than those in the model group(P<0.05);the positive expression rate of Ki67 in tumor tissue of mice in the low concentration group was lower than that in the medium concentration group(P<0.05);there was no significant difference in the positive expression rate of Ki67 in tumor tissue of mice between the high concentration group and the low concentration group,medium concentration group(P>0.05).The relative expressions of caspase-1,caspase-3,caspase-9 protein in tumor tissues of mice in low concentration group,medium concentration group and high concentration group were significantly higher than those in the model group(P<0.05).The relative expressions of caspase-1,caspase-3 protein in tumor tissues of mice in medium concentration group and high concentration group were significantly lower than those in the low concentration group(P<0.05);the relative expression of caspase-9 protein in tumor tissues of mice in high concentration group was significantly lower than that in the low concentration group(P<0.05);there was no statistical difference in the relative expression of caspase-9 protein in tumor tissues of mice between the medium concentration group and low concentration group(P>0.05).The relative expressions of caspase-3 protein in tumor tissues of mice in the high concentration group was significantly lower than that in the medium concentration group(P<0.05);there was no statistical difference in the relative expressions of caspase-1,caspase-9 protein in tumor tissues of mice between the high concentration group and medium concentration group(P>0.05).Conclusion Tubeimu can increase the daily food intake of the mice,inhibit the growth of the tumor,and reduce the expression of the tumor marker Ki67;it may inhibit tumor growth by up-regulating the expressions of caspase-1,caspase-3,caspase-9 protein.
作者
查青
陈燕妮
晁旭
ZHA Qing;CHEN Yanni;CHAO Xu(Shaanxi University of Chinese Medicine,Xi′an 712046,Shaanxi Province,China;Department of Science Research,the Second Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang 712000,Shaanxi Province,China)
出处
《新乡医学院学报》
CAS
2021年第4期301-307,共7页
Journal of Xinxiang Medical University
基金
国家自然科学基金资助项目(编号:81774132)
陕西中医药大学消化病肿瘤分子机制及中西医结合防治基础研究创新团队(编号:2019-YS05)。
关键词
土贝母
肝细胞癌
细胞凋亡
Tubeimu
hepatocellular carcinoma
apoptosis