蒙药红花清肝13味丸治疗CCl4诱导小鼠肠道损伤的转录组测序分析研究  

Transcriptome Sequencing Analysis Reveals the Possible Therapeutic Mechanisms of a Traditional Mongolian Medicine Honghua Qinggan 13 Flavor Pills in CCl4 Induced Intestinal Injury in Mice

作　　者：魏媛媛 红梅 王海生 WEI Yuanyuan;Hongmei;WANG Haisheng(Basic Medical College,Inner Mongolia Medical University,Hohhot010110,China)

机构地区：[1]内蒙古医科大学基础医学院,内蒙古呼和浩特010110

出　　处：《畜牧与饲料科学》2023年第6期13-21,共9页Animal Husbandry and Feed Science

基　　金：内蒙古自治区自然科学基金项目(2021BS08027);内蒙古自治区卫健委项目(202202143);内蒙古医科大学英才项目(YCPY2023100)。

摘　　要：[目的]筛选参与四氯化碳(CCl4)诱导的小鼠肠道损伤以及蒙药红花清肝13味丸治疗过程的重要信号通路及基因。[方法]将18只6~8周龄、体重为20~22 g的雄性C57BL/6小鼠随机分为3组,即空白对照组(n=6)、模型组(n=6)和治疗组(n=6)。空白对照组与模型组每天灌胃1次生理盐水,剂量为10 m L/(kg·BW),连续7 d;治疗组每天灌胃1次红花清肝13味丸,剂量为616.5 mg/(kg·BW),连续14 d;最后一次灌胃4 h后,空白对照组腹腔注射橄榄油,模型组和治疗组腹腔注射CCl4橄榄油混合物(CCl4∶橄榄油=1∶4,V/V),剂量均为10 m L/(kg·BW)。肠道损伤处理后第2天,分别采集3组小鼠十二指肠组织,进行高通量转录组测序。利用获得的测序数据,对差异表达基因进行预测和功能分析以及KEGG信号通路富集,筛选红花清肝13味丸治疗后显著下调信号通路富集的基因群,并与红花清肝13味丸药物靶点基因进行互交分析。[结果](1)转录组测序分析表明,与空白对照组相比,模型组与治疗组分别获得了1 419个和2 875个差异表达基因;与模型组相比,治疗组获得了2 288个差异表达基因,其中,上调表达基因1 139个,下调表达基因1 149个。对模型组差异表达基因进行KEGG信号通路富集,发现被激活的信号通路包括破骨细胞分化﹑B细胞﹑趋化因子﹑炎症性肠病﹑JAK-STAT等;对治疗组差异表达基因进行KEGG信号通路富集,发现被激活的信号通路包括FOXO﹑m TOR﹑长寿调节等,被抑制的信号通路包括破骨细胞分化﹑细胞凋亡﹑肿瘤坏死因子﹑B细胞﹑结肠直肠癌等。(2)筛选出治疗组显著抑制信号通路富集的基因群,与红花清肝13味丸药物靶点基因进行互交分析,进一步筛选出在转录组数据中表达量存在显著差异的4个基因,分别为JUN﹑STAT1﹑NFKBIA、FOS。[结论]CCl4诱导的小鼠肠道损伤以及蒙药红花清肝13味丸治疗过程涉及的主要信号通路包括趋化因子﹑炎症�[Objective]The aim of the present study was to screen the important signaling pathways and genes involved in carbon tetrachloride(CCl4)induced intestinal injury in mice and in the treatment process with a traditional Mongolian medicine Honghua Qinggan 13 Flavor Pills.[Method]A total of 18 male C57BL/6 mice aged 6-8 weeks and weighing 20-22 g were randomly divided into three groups,a blank control group(n=6),a model group(n=6),and a treatment group(n=6).The blank control group and model group were gavaged with physiological saline once a day at a dose of 10 mL/(kg·BW)for 7 consecutive days.The treatment group was gavaged with Honghua Qinggan 13 Flavor Pills once a day at a dose of 616.5 mg/(kg·BW)for 14 consecutive days.Four hours after the last gavage,the blank control group was intraperitoneally injected with olive oil,while both the model group and treatment group were intraperitoneally injected with CCl4 and olive oil mixture(CCl4∶olive oil=1∶4 V/V)at a dose of 10 mL/(kg·BW).On the second day following the intestinal injury treatment,the duodenal tissues of the mice in the three groups were collected for high-throughput transcriptome sequencing.Using the obtained sequencing data,the differentially expressed genes were predicted,functionally analyzed,as well as KEGG signaling pathway enriched.The gene groups enriched in the significantly down regulated signaling pathway after the treatment with Honghua Qinggan 13 Flavor Pills were screened,and interaction analysis was performed with the target genes of Honghua Qinggan 13 Flavor Pills.[Result]①Transcriptome sequencing analysis showed that compared with the blank control group,the model group and the treatment group obtained 1419 and 2875 differentially expressed genes,respectively.Compared with the model group,the treatment group obtained 2288 differentially expressed genes,of which 1139 were up regulated and 1149 were down regulated.KEGG signaling pathways enrichment on the differentially expressed genes in the model group found that the activated sign

关 键 词：肠道损伤 红花清肝13味丸 转录组测序 信号通路 差异基因 

分 类 号：R285.5[医药卫生—中药学] R-332[医药卫生—中医学]