畜牧兽医学报2024，Vol.55Issue(5)：2195-2205,11.DOI:10.11843/j.issn.0366-6964.2024.05.037
猪肺炎支原体乳酸脱氢酶在诱导猪支气管上皮细胞凋亡中的作用
Effects of LDH in Mesomycoplasma(Mycoplasma)hyopneumoniae on Apoptosis of Porcine Bronchial Epithelial Cells
摘要
Abstract
Mesomycoplasma(Mycoplasma)hyopneumoniae(Mhp)infection can cause damage and apoptosis of host epithelial cells,and the breakdown of the epithelial barrier often leads to the secondary infection of other bacteria and viruses,causing serious economic losses to the swine industry.However,the pathogenicity and virulence determinants of Mhp are not yet completely elucidated.Previous studies have found that Mhp lactate dehydrogenase(LDH)not only involved in the metabolism of Mhp,but also related to its virulence.Therefore,in this study,recombinant LDH(rLDH)was obtained through prokaryotic expression and affinity chromatography,and different concentrations of rLDH were used to interact with porcine bronchial epithelial cell(PBEC).The effect of rLDH on cellular morphology was observed by microscope,and the effect of rLDH on cellular viability was detected by CCK-8 method.The rLDH concentrations(50 and 100 μg·mL-1)were selected for further analysis by that there was no significant effect on cellular morphology and viability when it incubated with PBEC,and the apoptosis rate of cells was detected by flow cytometry.The transcriptional and expression levels of apoptosis-related genes caspase3,caspase 8,caspase 9,Bax and BCL-2 were detected by reverse transcription-quantitative polymerase chain reaction(RT-qPCR)and Western blot.The results showed that rLDH induced apoptosis of PBEC in a dose-dependent manner,the function of that was similar with Mhp.The mRNA levels of caspase 3,caspase 9,and Bax of PBEC after incubation with rLDH and Mhp 168 were significantly up-regulated and the mRNA level of BCL-2 was significantly down-regulated,while the mRNA of caspase 8 was not changed significantly.The protein expression levels of caspase 3 and Bax were significantly up-regulated and the protein expression level of BCL-2 was significantly down-regulated.The results showed that Mhp could induce the endogenous apoptosis of PBEC through LDH,in which the apoptosis-related factors caspase 3,caspase 9,Bax and BCL-2 playing important roles.关键词
猪肺炎支原体(Mhp)/乳酸脱氢酶(LDH)/猪支气管上皮细胞(PBEC)/凋亡Key words
Mesomycoplasma hyopneumoniae/lactate dehydrogenase/PBEC/apoptosis分类
农业科技引用本文复制引用
王吉英,于岩飞,尹蕊如,谢星,王海燕,刘胡栋,胡辉,熊祺琰,冯志新,邵国青..猪肺炎支原体乳酸脱氢酶在诱导猪支气管上皮细胞凋亡中的作用[J].畜牧兽医学报,2024,55(5):2195-2205,11.基金项目
国家重点研发计划(2022YFD1800903) （2022YFD1800903）
湖南省教育厅科学研究项目(22B1055) （22B1055）
湖南省自然科学基金项目(2022JJ50321) （2022JJ50321）
国家自然科学基金项目(32172860 （）
32273011) （）
江苏省科协青年科技人才托举工程(JSTJ-2023-XH031) （JSTJ-2023-XH031）
