摘要
目的探讨氧化苦参碱对结直肠癌小鼠肠道菌群的影响及其对小鼠结直肠癌作用的相关微生物机制。方法将16只5周龄BALB/c雄性小鼠通过氧化偶氮甲烷(AOM)-葡聚糖硫酸钠(DSS)法建立小鼠原位结直肠肿瘤模型,采用分层抽样的方法分为对照组和氧化苦参碱干预组,每组8只;其中氧化苦参碱干预组于造模第5周开始接受10 mg/kg氧化苦参碱溶液隔日腹腔注射,对照组接受等量0.9%NaCl注射液腹腔注射,至造模第81天实验结束。自开始造模,每3 d测量1次小鼠体质量。于处死前收集小鼠粪便,进行微生物及16S核糖体DNA(rDNA)高通量测序。观察结直肠肿瘤数,取肿瘤组织,采用苏木精-伊红(HE)染色判断肿瘤分化程度,计算各分化级别肿瘤组织灶占总肿瘤组织灶的比例;免疫组织化学法检测Ki-67表达情况。结果实验后期(造模第49天至第81天),对照组小鼠体质量低于氧化苦参碱干预组[造模第81天体质量:(22.9±0.5)g比(24.0±0.5)g,t=2.187,P<0.05],实验结束时,氧化苦参碱干预组肠道肿瘤数少于对照组[(8.5±1.2)个比(12.0±1.2)个,t=2.824,P<0.05];氧化苦参碱干预组小鼠肠道高分化肿瘤灶占比高于对照组[(62.5±3.7)%比(25.0±2.6)%],Ki-67表达评分低于对照组[(3.2±1.0)分比(6.0±1.0)分,t=2.668,P<0.05]。在门水平,氧化苦参碱干预组小鼠肠道中厚壁菌门、变形菌门丰度高于对照组(均P<0.05),拟杆菌门丰度低于对照组(P=0.037)。在属水平,氧化苦参碱干预组norank_f__桑科菌丰度高于对照组(P=0.001),拟杆菌属、臭气杆菌属、副细菌属、异波菌属丰度低于对照组(均P<0.05)。结论氧化苦参碱可能通过调节小鼠肠道菌群维持肠道菌群稳定,从而降低肠道肿瘤的发生率,延缓肿瘤进展。
Objective To explore the effect of oxymatrine on the intestinal flora of mice with colorectal cancer and its related microbial mechanisms.Method Based on azoxymethane(AOM)-dextran sulfate sodium(DSS)method,165-week-old male BALB/c mice were performed to establish a orthotopic colorectal tumor mouse model.According to the stratified sampling method,mice were divided into the control group and oxymatrine intervention group,8 in each group.From 5th week,mice in oxymatrine intervention group were given intraperitoneal injection of 10 mg/kg oxymatrine solution every other day;mice in the control group were given the same amount of 0.9%NaCl injection intraperitoneally until the end of the experiment at 81st d of modeling.The body weight of mice was measured every 3 days since the beginning of the modeling;before mice were sacrificed,mouse feces were collected for microbiological and 16S ribosome(rDNA)highthroughput sequencing.The tumor number of colorectal cancer was observed and tumor tissues were taken out.Hematoxylin and eosin(HE)staining was used to evaluate the differentiation degree,the percentage of tumor tissues with all differentiation degrees in the total tumor tissues was calculated,and immunohistochemistry staining was used to test the expression of Ki-67.Results At the late of the experiment(d 49-d 81 of modeling),the body weight of mice in the control group was lower than that of mice in oxymatrine intervention group[modeling at 81st d:(22.9±0.5)g vs.(24.0±0.5)g,t=2.187,P<0.05],and the tumor number of intestinal tract in oxymatrine intervention group was lower than that in the control group[(8.5±1.2)vs.(12.0±1.2),t=2.824,P<0.05]at the end of experiment.The percentage of well-differentiated tumors in mice intestinal tract in the oxymatrine intervention group was higher than that of mice in the control group[(62.5±3.7)%vs.(25.0±2.6)%],and the expression score of Ki-67 in oxymatrine intervention group was lower compared with that in the control group[(3.2±1.0)scores vs.(6.0±1.0)scores,t=2.668,P<0.05).At the phylum level,the abundance of Firmicutes and Proteobacteria in the intestine of mice in oxymatrine intervention group was higher than that in the control group(all P<0.05),the abundance of Bacteroides in oxymatrine intervention group was lower than that in the control group(P=0.037).At the genus level,compared with the control group,the oxymatrine intervention group had a higher abundance of norank_f__Muribaculaceae(P=0.001).The abundance of Bacteroides,Odoribacter,Parabacteroides and alloprevotella in oxymatrine intervention group was lower than that in the control group(all P<0.05).Conclusion Oxymatrine can decrease the incidence of colorectal cancer in mice and delay development of colorectal cancer by regulating the gut microbiota and sustaining the stability of intestinal flora.
作者
高珊
柳清
Gao Shan;Liu Qing(Department of Pathology,the Second Affiliated Hospital of Harbin Medical University,Harbin 150086,China)
出处
《肿瘤研究与临床》
CAS
2021年第9期645-650,共6页
Cancer Research and Clinic
基金
国家自然科学基金(8170110698)。
关键词
结直肠肿瘤
模型
动物
病理评价
肠道菌群
Colorectal neoplasms
Models,animal
Pathological evaluation
Gut bacteria
作者简介
通信作者:柳清,Email:liuqingsix1217@126.com。
