摘要
为研究苦参碱对A型产气荚膜梭菌致小鼠肠炎的预防和治疗效果,本实验将56只8周龄的健康雄性昆明小鼠均分为对照组、菌液组、治疗组和预防组。菌液组小鼠以0.2 mL/只灌胃A型产气荚膜梭菌(1×10~9cfu/mL)16 d,治疗组小鼠于灌胃A型产气荚膜梭菌7 d后灌服苦参碱(30 mg/kg)9 d,预防组小鼠则以A型产气荚膜梭菌菌液和苦参碱同时灌胃16 d,对照组小鼠则每日每只灌服PBS 0.2 mL至16 d。对各组小鼠称重采血后剖杀、收集回肠组织制备病理切片观察组织病变,并分离血清,利用ELISA试剂盒对血清中的炎性细胞因子进行检测,采集肠内容物,经PCR检测后灰度分析A型产气荚膜梭菌的数量,采用ELISA试剂盒测定α毒素含量。结果显示治疗组和预防组小鼠的体质量明显增加;菌液组小鼠肠道小肠绒毛脱落,淋巴细胞和红细胞的数量增加,而预防组和治疗组的肠道炎性细胞浸润减少,肠绒毛结构逐渐恢复,炎症引起的肠组织损伤得以修复。ELISA检测结果显示,预防组小鼠血清中IL-1β、IL-6、TNF-α、MAPK8的含量与对照组之间无显著性差异(P>0.05);治疗组小鼠血清中IL-6和MAPK8含量显著高于对照组(P<0.05)。PCR检测后灰度分析结果显示,治疗组和预防组小鼠肠内容物A型产气荚膜梭菌的数量与菌液组无显著差异(P>0.05),但与对照组比较,上述两组小鼠释放的A型产气荚膜梭菌α毒素含量明显降低。上述结果表明苦参碱可抑制A型产气荚膜梭菌释放ɑ-毒素,并可恢复小鼠回肠黏膜免疫屏障,降低荚膜梭菌所引起小鼠的肠道炎症。本研究为苦参碱应用于A型产气荚膜梭菌所致肠炎的治疗和预防提供参考依据。
To investigate the preventive and therapeutic effects of matrine (MT) on diarrhea induced by Clostridium perfringens type A in mice,56 healthy 8-week-old male Kunming mice were evenly divided into four groups:control group (PBS),bacterial suspension group (Cp A 0.2mL/mouse,1×10~9cfu/mL),treatment group,and prevention group.In the treatment group,mice were intragastric administered MT (30mg/kg) for 9 days after Clostridium perfringens type A gavage for 7 days.In the prevention group mice were simultaneously intragastric administered with Clostridium perfringens type A bacterial suspension and MT for 16 days After weighing the mice and collecting blood in each group,serum was separated,and inflammatory cytokines in the serum were detected using the ELISA method.Ileal tissues and intestinal contents were collected to determine the number of Clostridium perfringens type A by PCR and the amount of α-toxin released by ELISA kit.The results showed that the body weight of mice in the treatment and prevention groups increased significantly.In the bacterial suspension group,the intestinal villi of the mice were shed,and the number of lymphocytes and red blood cells increased.However,in the prevention and treatment groups,the infiltration of inflammatory cells in the intestine decreased,the structure of intestinal villi gradually recovered,and the intestinal tissue damage caused by inflammation was repaired.ELISA results showed no significant difference in the levels of IL-1β,IL-6,TNF-α,and MAPK8 in the serum of mice in the pre-vention group compared with the control group (P>0.05).The levels of IL-6 and MAPK8 in the treatment group were significantly higher than those in the control group (P<0.05).PCR results showed no significant difference in the amount of Clostridium perfringens type A in the intestinal contents of mice in the treatment and prevention groups compared with the bacterial suspension group (P>0.05).However,compared with the control group,there was a significant inhibitory effect on the content of α-toxin produced by Clostridium perfringens type A.These results indicate that MT can inhibit the release of α-toxin by Clostridium perfringens type A,restore the mucosal immune barrier of the ileum in mice,and reduce intestinal inflammation caused by Clostridium perfringens.This study provides a theoretical basis for the application of MT in the treatment and prevention of enteritis caused by Clostridium perfringens type A.
作者
李慧
李晓林
秦颖
孟妮佳
吴晨晨
刘文君
薛虹
LI Hui;LI Xiao-lin;QING Ying;MENG Ni-jia;WU Chen-chen;LIU Wen-jun;XUE Hong(Harbin Sport University,Harbin 150001,China;Northwest A&F University,Xianyang 712199,China)
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2024年第7期740-746,共7页
Chinese Journal of Preventive Veterinary Medicine
基金
黑龙江省高校基本科研业务(2023 KYYWF-PY10)。
关键词
苦参碱
A型产气荚膜梭菌
小鼠
体外抑制
治疗与预防
Matrine
Clostridium perfringens type A
mice
inhibition in vitro
treatment and prevention
