梅花草提取物调节肠道菌群抑制原位脑胶质瘤模型的作用机制研究  

Study on the mechanism of Meihuacao extracts regulating gut microbiota to suppress primary glioma models

作　　者：王睿君[1] 马宇衡 王儒帅 何占彪[4] WANG Ruijun;MA Yuheng;WANG Rushuai;HE Zhanbiao(Inner Mongolia Clinical Medical Research Center of Nervous System Diseases,Department of Neurosurgery,Affiliated Hospital,Inner Mongolia Medical University,Hohhot 010050,China;College of Pharmacy,Inner Mongolia Medical University,Hohhot 010050,China;Department of Oncology,Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010050,China;Department of Neurosurgery,Affiliated Hospital,Inner Mongolia Medical University,Hohhot 010050,China)

机构地区：[1]内蒙古自治区神经系统疾病临床医学研究中心·内蒙古医科大学附属医院神经外科,内蒙古自治区呼和浩特010050 [2]内蒙古医科大学药学院,内蒙古自治区呼和浩特010050 [3]内蒙古医科大学附属医院肿瘤内科,内蒙古自治区呼和浩特010050 [4]内蒙古医科大学附属医院神经外科,内蒙古自治区呼和浩特010050

出　　处：《中国实验诊断学》2024年第10期1205-1213,共9页Chinese Journal of Laboratory Diagnosis

基　　金：国家自然科学基金项目(82060906);内蒙古自治区科技厅自然科学基金项目(2020MS08146);内蒙古自治区卫健委医疗卫生科技计划项目(202201307);内蒙古医科大学重点项目(YKD2021ZD002);内蒙古医科大学附属医院院级项目——科研联合基金(2023NYFYLHZD007)。

摘　　要：目的探讨脑胶质瘤发生过程中梅花草对原位荷瘤鼠肠道菌群的影响。方法4周龄Balb/c雌性裸鼠颅内种植U87-MG或者RFP-U87-MG脑胶质瘤,前者用于肠道菌群检测(每组6只,分为2组),后者用于活体荧光成像评价脑胶质瘤的生长状态(每组3只,分为2组)。梅花草提取物经灌胃(25 mg/kg)隔天给药连续10次(植入后第6天至第24天),对照组为1×PBS。RFP-U87-MG模型鼠在第14天和第24天进行活体荧光成像观察肿瘤生长状态,同时进行裸鼠称量,并在第25天对裸鼠实施安乐死后,剥离脑组织和肿瘤,分别进行称重和组织的HE染色;而种植U87-MG的模型鼠,每3天称重,第25天实行安乐死收集结肠内新鲜粪便样本,以Illumina高通量测序技术分析肠道菌群,进行生物信息学挖掘潜在梅花草治疗相关的特征菌群。结果活体荧光成像和肿瘤病灶称重均提示梅花草提取物显著抑制原位脑肿瘤的恶性增殖,HE染色显示梅花草组肿瘤细胞数量减少、核浆比趋于正常,核异型性降低。经高通量测序发现梅花草提取物干预后并未影响荷瘤鼠肠道菌群的α多样性,而是影响特征菌群的丰度,其中梅花草提取物治疗后可以显著提高Muribaculum intestinale、Bacteroides faecichinchillae、Klebsiella oxytoca的菌种丰度,降低Lachnospiraceae bacterium A4和Helicobacter japonicus的丰度。结论梅花草提取物在显著抑制原位脑胶质瘤恶性增殖的同时,影响肠道菌群的丰度,其中特征菌群如Muribaculum intestinale、Bacteroides faecichinchillae等丰度增加,可能作为潜在益生菌参与脑肿瘤的机制调控。Objective To investigate the effect of Meihuacao on intestinal microbiota of bearing mice during glioma development.Methods The 4-week-old Balb/c female nude mice were intracranially implanted with either U87-MG or RFP-U87-MG gliomas,the former for intestinal flora assay(6per group,divided into 2groups),and the latter for in vivo fluorescence imaging to evaluate the growth status of gliomas(3per group,divided into 2groups).Meihuacao extracts were administered by gavage(25mg/kg)every other day for 10consecutive times(from day 6to day 24after implantation),and the control group was 1*PBS.The RFP-U87-MG model mice were subjected to in vivo fluorescence imaging to observe the tumor growth status on days 14and 24,while nude mice were weighed,and after euthanasia of nude mice on day 25,the brain tissues and tumors were exfoliated and subjected to separate weighing following HE staining of these tissues.While the U87-MG model mice were weighed each 3days,fresh fecal specimens were collected from the colon by euthanasia on the 25th day,and the gut microbes was analyzed by Illimina high-throughput sequencing for bioinformatics mining of characteristic species potentially related to the treatment of Meihuacao.Results In vivo fluorescence imaging and weighing of tumor foci suggested that Meihuacao extracts significantly inhibited the malignant proliferation of primary gliomas,and HE staining exhibited that the number of tumor cells in the Meihuacao group was reduced,the nuclear-to-plasma ratio tended to be normalized,and the nuclear anisotropy was reduced.After highthroughput sequencing,it was found that the intervention of Meihuacao extracts did not affect theα-diversity of the intestinal flora,but rather the abundance of the characteristic microbes,in which the treatment of Meihuacao extracts could significantly increase Muribaculum intestinale,Bacteroides faecichinchillae,Klebsiella oxytoca,while decreasing the abundance of Lachnospiraceae bacterium A4and Helicobacter japonicus.Conclusion While significantly inhibiting

关 键 词：脑胶质瘤 梅花草提取物 肠道微生态 脑-肠轴 荧光成像 

分 类 号：R739.41[医药卫生—肿瘤]