枯草芽孢杆菌培养上清液对绵羊瘤胃上皮细胞β-防御素-1表达的影响  

Effect of Bacillus subtilis Culture Supernatant onβ-defensin-1 Expression in Ovine Ruminal Epithelial Cells

作　　者：白绥明 辛雅明 齐盟 胡亮 杨银凤[1] BAI Suiming;XIN Yaming;QI Meng;HU Liang;YANG Yinfeng(College of Veterinary Medicine,Inner Mongolia Agricultural University/Key Laboratory of Clinical Diagnosis and Treatment Technology for Animal Diseases,Minister of Agriculture and Rural Affairs,Hohhot 010018,China;Center for Animal Disease Prevention and Control of Qingjian County,Yulin 718399,China;Center for Animal Disease Prevention and Control of Ordos City,Ordos 017000,China;Bayannur Institute of Agricultural and Animal Husbandry Sciences,Bayannur 015000,China)

机构地区：[1]内蒙古农业大学兽医学院/农业农村部动物疾病临床诊疗技术重点实验室,内蒙古呼和浩特010018 [2]清涧县动物疫病预防控制中心,陕西榆林718399 [3]鄂尔多斯市动物疫病预防控制中心,内蒙古鄂尔多斯017000 [4]巴彦淖尔市农牧业科学研究所,内蒙古巴彦淖尔015000

出　　处：《畜牧与饲料科学》2023年第5期8-14,共7页Animal Husbandry and Feed Science

基　　金：国家自然科学基金项目(32060780)。

摘　　要：[目的]探究枯草芽孢杆菌培养上清液对绵羊瘤胃上皮细胞(ovine ruminal epithelial cells,ORECs)中绵羊β-防御素-1(sheepβ-defesin-1,SBD-1)表达的影响。[方法]采用Cell Counting kit-8(CCK-8)方法筛选对ORECs无明显毒性作用的枯草芽孢杆菌培养上清液刺激浓度范围与刺激时间范围。用筛选出的无明显毒性作用的不同浓度枯草芽孢杆菌对应的培养上清液刺激ORECs 8 h后,采用荧光定量PCR(qPCR)技术和酶联免疫吸附(ELISA)检测方法筛选枯草芽孢杆菌培养上清液诱导SBD-1表达的最佳刺激浓度,以该浓度枯草芽孢杆菌对应的培养上清液刺激ORECs不同时间,采用qPCR技术和ELISA检测方法确定枯草芽孢杆菌培养上清液诱导SBD-1表达的最佳刺激时间。[结果]①对ORECs无明显毒性作用的枯草芽孢杆菌对应的培养上清液浓度范围为10^(7)~10^(11) CFU/mL,刺激时间范围为2~24 h。②利用qPCR技术和ELISA检测方法筛选出最佳刺激浓度为10^(11) CFU/mL枯草芽孢杆菌对应的培养上清液。③以该浓度枯草芽孢杆菌对应的培养上清液刺激ORECs不同时间,采用qPCR技术和ELISA检测方法确定枯草芽孢杆菌培养上清液诱导SBD-1表达的最佳刺激时间为24 h。[结论]枯草芽孢杆菌培养上清液可以诱导ORECs SBD-1的表达。[Objective]This study was conducted to assess the effect of Bacillus subtilis culture supernatant on sheepβ-defesin-1(SBD-1)expression in ovine ruminal epithelial cells(ORECs).[Method]The corresponding supernatants of Bacillus subtilis cultures that had no obvious toxic effects on ORECs were screened using the cell counting kit-8(CCK-8)assay to determine the ranges of proper stimulating concentration and time.After stimulating ORECs with the different concentrations of Bacillus subtilis culture supernatants that had no obvious toxic effects for 8 h,the optimal stimulating concentration for inducing SBD-1 expression was determined by using qPCR and ELISA assays.In addition,this concentration of Bacillus subtilis culture supernatant was subsequently used to stimulate ORECs for various durations,and the optimal stimulating time for inducing SBD-1 expression was determined by using qPCR and ELISA assays.[Result]①The corresponding supernatants of Bacillus subtilis cultures at concentrations ranging from 10^(7) to 10^(11) CFU/mL exhibited no obvious toxic effects on ORECs,and the range of proper stimulating time was 2-24 h.②The best performance in inducing SBD-1 expression was observed in the corresponding supernatant of Bacillus subtilis cultures at a concentration of 10^(11) CFU/mL.③After treatment with the screened optimal stimulating concentration of Bacillus subtilis culture supernatant,the optimal stimulating time for inducing SBD-1 expression was determined as 24 h.[Conclusion]Bacillus subtilis culture supernatant could induce the SBD-1 expression in ORECs.

关 键 词：绵羊瘤胃上皮细胞 枯草芽孢杆菌培养上清液 β-防御素-1 荧光定量PCR 

分 类 号：S858.262.4[农业科学—临床兽医学]