肉桂酸-小檗碱共晶长期给药对小鼠肠道屏障及肠道菌群的影响  

Effect of Long-term Administration of Cinnamic Acid-Berberine Cocrystal on Intestinal Barrier and Intestinal Flora in Mice

作　　者：高文恒 李孟孟 何勇 何仔昂 杨晔 尹登科 GAO Wenheng;LI Mengmeng;HE Yong;HE Ziang;YANG Ye;YIN Dengke(School of Pharmacy,Anhui University of Chinese Medicine,Anhui Hefei 230012,China;School of Chemistry and Chemical Engineering,Hefei University of Technology,Anhui Hefei 230009,China;Anhui Province Key Laboratory of Pharmaceutical Preparation Technology and Application,Anhui Hefei 230012,China;Anhui Province Key Laboratory of Chinese Medicinal Formula,Anhui Hefei 230012,China)

机构地区：[1]安徽中医药大学药学院,安徽合肥230012 [2]合肥工业大学化学与化工学院,安徽合肥230009 [3]药物制剂技术与应用安徽省重点实验室,安徽合肥230012 [4]中药复方安徽省重点实验室,安徽合肥230012

出　　处：《安徽中医药大学学报》2024年第5期59-66,共8页Journal of Anhui University of Chinese Medicine

基　　金：安徽省高校重大科研项目(2022AH040077);安徽省高校学科(专业)拔尖人才学术资助项目(gxbjZD2021056);安徽省高校科研计划团队项目(2022AH010036)。

摘　　要：目的观察肉桂酸—小檗碱共晶(cinnamic acid-berberine cocrystal,CBBR)和小檗碱(berberine,BBR)长期口服给药对小鼠肠道屏障及肠道菌群的影响。方法将小鼠随机分为对照(control,CTL)组、BBR组和CBBR组,每组6只;给药组小鼠每天灌胃相应药物(BBR:70 mg/kg;CBBR:100 mg/kg),连续灌胃9周。每周称量小鼠体质量,9周后麻醉处死小鼠,测定小鼠结肠长度;HE染色观察小鼠结肠组织病理变化;AB-PAS染色观察小鼠结肠黏液层厚度和杯状细胞数量变化;免疫荧光染色法和Western blot法检测小鼠结肠紧密连接蛋白(ZO-1、Claudin-1、Occludin)表达水平;16S rDNA测序分析小鼠肠道菌群变化。结果与CTL组比较,BBR组和CBBR组小鼠体质量显著减少(P<0.05);各组小鼠结肠长度比较,差异无统计学意义(P>0.05);HE和AB-PAS染色结果显示,小鼠结肠组织结构、杯状细胞及黏液量无明显改变;免疫荧光染色和Western blot法检测结果显示,与对照组比较,给药组ZO-1、Claudin-1、Occludin蛋白表达差异无统计学意义(P>0.05);α多样性分析和主坐标分析显示,BBR和CBBR改变肠道菌群的多样性和结构组成,显著升高另枝菌属(Alistipes)、臭杆菌属(Odoribacter)、罗氏菌属(Roseburia)、瘤胃球菌属(norank_f_Ruminococcaceae)、优/真杆菌属(Eubacterium_xylanophilum_group)的丰度(P<0.05);与BBR组比较,CBBR组臭杆菌属(Odoribacter)、优/真杆菌属(Eubacterium_xylanophilum_group)丰度显著升高(P<0.05),拟杆菌属(Bacteroides)丰度显著下降(P<0.05)。结论CBBR和BBR长期给药可显著降低小鼠体质量,能调节小鼠肠道菌群的组成和某些菌属的丰度,对小鼠肠道屏障无显著影响。Objective To investigate the effect of long-term oral administration of cinnamic acid-berberine cocrystal(CBBR)versus berberine(BBR)on intestinal barrier and intestinal flora in mice.Methods Mice were randomly divided into control group(CTL group),BBR group(70 mg/kg),and CBBR group(100 mg/kg),and the drugs were given by gavage for 9 consecutive weeks.Body weight of the mice was measured every week,and after 9 weeks,the mice were anesthetized and sacrificed to measure the length of colon.HE staining was used to observe the histopathological changes of the colon;AB-PAS staining was used to measure the thickness of the colonic mucus layer and the number of goblet cells;immunofluorescent staining and Western blot were used to measure the expression levels of the tight junction proteins zonula occluden-1(ZO-1),Claudin-1,and Occludin in the colon;16S rDNA sequencing was used to analyze the change in intestinal flora.Results Compared with the CTL group,the BBR group and the CBBR group had a significant reduction in body weight(P<0.05).There was no significant difference in the length of the colon between the groups(P>0.05).HE and AB-PAS staining showed no obvious changes in colon structure,goblet cells,and mucus layer.Immunofluorescent staining and Western blot showed no significant differences in the expression of the tight junction proteins ZO-1,Claudin-1,and Occludin between the administration groups and the control group(P>0.05).The alpha-diversity analysis and the principal coordinate analysis showed alterations in the abundance and structural composition of intestinal flora after the administration of BBR and CBBR,with significant increases in the abundance of Alistipes,Odoribacter,Roseburia,norank_f_Ruminococcaceae,and Eubacterium_xylanophilum_group(P<0.05),and compared with the BBR group,the CBBR group had significant increases in the abundance of Odoribacter and Eubacterium_xylanophilum_group(P<0.05)and a significant reduction in the abundance of Bacteroides(P<0.05).Conclusion The long-term administration of CB

关 键 词：小檗碱 肉桂酸—小檗碱共晶 肠道屏障 肠道菌群 

分 类 号：R289.5[医药卫生—方剂学] R255.6[医药卫生—中药学]