自噬与蛋白酶体途径在布鲁菌感染巨噬细胞过程中的相互作用  

Interaction of autophagy and ubiquitin proteasome system after macrophage infected Brucella

作　　者：董炳梅 梁萌萌 许文雅 王君 宋立立 任洪林 DONG Bingmei;LIANG Mengmeng;XU Wenya;WANG Jun;SONG Lili;REN Honglin(College of Life Science,Cangzhou Normal University,Hebei 061001,China;Key Laboratory of Zoonosis Research of Ministry of Education/National Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases,College of Veterinary Medicine/Institute of Zoonosis,Jilin University,Changchun 130062,China)

机构地区：[1]沧州师范学院生命科学学院,河北沧州061001 [2]吉林大学动物医学学院/人兽共患病研究所人畜共患传染病重症诊治全国重点实验室/人兽共患病研究教育部重点实验室,吉林长春130062

出　　处：《中国兽医学报》2025年第3期507-513,共7页Chinese Journal of Veterinary Science

基　　金：沧州师范学院科研创新团队资助项目(cxtdl2303);山东省自然科学基金资助项目(ZR2013CQ005)。

摘　　要：为了探明巨噬细胞(macrophage,M_(φ))感染布鲁菌(Brucella)后泛素-蛋白酶体途径(ubiquitin proteasome system,UPS)和自噬-溶酶体途径(autophagy-lysosome pathway,ALP)在对抗Brucella过程中的相互作用关系,一方面将猪种布鲁菌(Brucella suis,B.suis)分别体外感染M_(φ)和腹腔接种BALB/c小鼠,并对细胞上清液和腹腔液中LC3Ⅱ、P62与20S蛋白酶体水平进行了测定,结果显示B.suis感染过程中UPS首先被激活,随后ALP途径发挥作用;另一方面,应用3-MA、乳胞素分别制备ALP抑制态M_(φ)与UPS抑制态M_(φ)并接种B.suis,随后对细胞上清中LC3Ⅱ、P62与20S蛋白酶体水平进行测定,结果显示当M_(φ)UPS功能被抑制时,ALP作用被显著提高;而ALP功能被抑制时,UPS活性未有显著变化。最后,将不同状态M_(φ)接种B.suis后,对B.suis的胞内增殖能力进行测定,结果显示当UPS与ALP功能被同时抑制时,B.suis的胞内增殖能力显著降低,且UPS对B.suis胞内增殖的促进作用强于ALP。结果表明,Brucella感染M_(φ)过程中UPS与ALP存在相互作用关系,在理论上可为Brucella与宿主细胞相互作用机制的研究提供新思路。In order to explore the relationship between ubiquitin proteasome system(UPS)and autophagy after macrophage was infected with Brucella.On the one hand,the levels of LC3Ⅱ、P62and 20S proteasomes in cell supernatant and peritoneal fluid were determined respectively after RAW264.7 cells were infected and BALB/c mice were intraperitoneal inoculated with Brucella suis(B.suis).The results displayed that UPS was activated firstly,followed by autophagy after B.suis infection.On the other hand,we prepared the UPS-inhibited-cells and ALP-inhibited-cells by lactacystin and 3-methy ladenine(3-MA),and then the cells were infected by B.suis and the levels of IC3Ⅱ、P62与20S proteasomes in cell supernatant were determined.The results showed that the ALP function was significantly improved when the effect of M_(φ)UPS was inhibited.However,when ALP is inhibited,UPS function is not improved effectively.At the last,the UPS and ALP co-inhibitory cells were prepared and the intracellular proliferation capacity of B.suis was determined.The result displayed that the intracellular proliferation capacity of B.suis was significantly reduced,and the promotion effect of UPS on the intracellular proliferation of B.suis was stronger than that of ALP.All above,this study preliminarily identified the interaction between UPS and ALP after M_(φ)infected Brucella,which theoretically provides a new idea for the study on interaction mechanism between Brucella and host cells.

关 键 词：自体吞噬 泛素蛋白酶体途径 布鲁菌 巨噬细胞 

分 类 号：S852.3[农业科学—基础兽医学] S852.61[农业科学—兽医学]