柴芪益肝颗粒改善H22肝癌荷瘤小鼠肠道黏膜通透性调控菌群的抗肝癌作用机制研究  

Study on the anti-hepatocellular carcinoma mechanism of H22 liver cancer-bearing mice of Chaiqi Yigan granule in improving intestinal mucosal permeability regulation flora

作　　者：庞旭 李晓斌 孙童 刘博文 胡世平 PANG Xu;LI Xiao-bin;SUN Tong;LIU Bo-wen;HU Shi-ping(Department of Hepatology,Dongzhimen Hospital,Beijing University of Chinese Medicine,Bejing 100700,China;Department of Hepatology,Shenzhen Hospital,Bejing University of Chinese Medicine,Shenzhen 518172,Guangdong Province,China)

机构地区：[1]北京中医药大学东直门医院肝病科,北京100700 [2]北京中医药大学深圳医院(龙岗)肝病科,广东深圳518172

出　　处：《中国临床药理学杂志》2024年第22期3301-3305,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金面上基金资助项目(81973733);深圳市科技计划基金资助项目(JCYJ20220530172812028);深圳市龙岗医疗卫生科技创新计划基金资助项目(LGKCYLWS2021000015);深圳市龙岗区医疗卫生技术攻关基金资助项目(LGKCYLWS2022006)。

摘　　要：目的研究柴芪益肝颗粒对H22肝癌荷瘤小鼠的治疗作用及其对肠道菌群的影响。方法用H22细胞悬液皮下接种方式构建肝癌小鼠模型。将小鼠随机分为正常组、模型组(肝癌模型)、阳性对照组(50mg·kg^(-1)索拉非尼)和高、低剂量实验组(49.68和12.42g·kg^(-1)柴芪益肝颗粒)。剥离瘤体,瘤体称质量,测定抑瘤率;用酶联免疫吸附试验(ELISA)法检测肝癌血清标志物甲胎蛋白(AFP);用16srRNA检测肠道菌群结构;用免疫组织化学染色法观察结肠黏膜中紧密连接蛋白闭锁因子(Occludin)和紧密连接蛋白-1(ZO-1)的表达及肠道黏膜通透性的改变。结果模型组、低剂量实验组、高剂量实验组和阳性对照组瘤体质量分别为(1.18±0.08)、(1.13±0.10)、(0.84±0.13)和(0.55±0.07)g,AFP的表达水平分别为(7.93±0.21)、(7.74±0.23)、(7.25±0.53)和(5.59±0.03)ng·mL^(-1),高剂量实验组的上述指标与模型组比较,在统计学上差异均有统计学意义(均P<0.01)。正常组、模型组、低剂量实验组、高剂量实验组和阳性对照组的拟杆菌门分别为(32.10±8.67)%、(12.72±4.31)%、(50.34±13.48)%、(28.37±6.17)%和(44.47±2.85)%,厚壁菌门分别为(67.18±4.20)%、(82.47±4.64)%、(38.16±12.88)%、(68.39±5.83)%和(52.61±2.86)%,乳杆菌科分别为(12.20±3.25)%、(1.85±0.63)%、(9.17±2.47)%、(3.56±0.74)%和(27.83±6.77)%,高剂量实验组的上述指标与模型组相比,在统计学上差异均有统计学意义(P<0.01,P<0.05)。结肠免疫组化结果显示,模型组的细胞排列松散,细胞间蛋白减少,细胞间隙增大,导致黏膜的通透性增加。随着柴芪益肝颗粒剂量的增加,Occludin蛋白和ZO-1蛋白逐渐增加,使得细胞排列整齐有序,细胞间隙变小,黏膜的通透性逐渐向正常恢复。结论柴芪益肝颗粒可能通过改善肠道黏膜通透性,调控肠道菌群发挥抗肝癌作用。Objective To study the therapeutic effect of Chaiqi Yigan granules on H22 liver cancer-bearing mice and its effect on intestinal flora.MethodsSThe mouse model of liver cancer was constructed by subcutaneous inoculation of H22 cell suspension.The rats were divided into normal group,model group(H22 liver cancer-bearing model),positive control group(50 mg·kg^(-1) Sorafenib)and experimental-L,-H groups(12.42,49.68 g·kg^(-1) Chaiqi Yigan granules).Stripping the tumor,weighing the tumor.Thee serum marker alpha-fetal protein(AFP)of liver cancer was detected by enzyme-linked immunosorbent assay(ELISA)method.The intestinal flora was detected by 16 s rRNA.The expression of tight junction proteins Occludin and zonula occludens protein-1(ZO-1)in colonic mucosa and the changes of intestinal mucosal permeability were observed by immunohistochemical staining.Results The tumor weights in model group,experimental-L group,experimental-H group and positive control group were(1.18±0.08),(1.13±0.10),(0.84±0.13)and(0.55±0.07)g;the expression of AFP were(7.93±0.21),(7.74±0.23),(7.25±0.53)and(5.59±0.03)ng mL^(-1),respectively.There were significant differences in the above indexes between the experimental-H group and the model group(P<0.01).The Bacteroidota in normal group,model group,experimental-L group,experimental-H group and positive control group were(32.10±8.67)%,(12.72±4.31)%,(50.34±13.48)%,(28.37±6.17)%and(44.47±2.85)%;Firmicutes were(67.18±4.20)%,(82.47±4.64)%,(38.16±12.88)%,(68.39±5.83)%and(52.61±2.86)%;Lactobacillaceae were(12.20±3.25)%,(1.85±0.63)%,(9.17±2.47)%,(3.56±0.74)%and(27.83±6.77)%,respectively.There were significant differences in the above indexes between the experimental-H group and the model group(P<0.01,P<0.05).The results of colonic immunohistochemistry showed that the cells in the model group were loosely arranged,the intercellular proteins were reduced,and the intercellular space was increased,resulting in increased mucosal permeability.With the increase of the dose of Chaiqi Yigan g

关 键 词：柴芪益肝颗粒 肝癌 肠道菌群 肠道黏膜通透性 

分 类 号：R28[医药卫生—中药学]