CAJ | 학술논문

目的观察AIM2( absent in melanoma 2)在感染早期识别胞浆小鼠巨细胞病毒(MCMV) DNA的变化状况.方法建立MCMV全身播散型感染模型,接种MCMV Smith株后第1、3、5和7天各处死3只小鼠；同时设模拟感染小鼠作为正常对照.Western blot检测脾脏巨噬细胞中AIM2、衔接子凋亡相关点样蛋白(ASC)和caspase-1蛋白的表达状况；同时应用ELISA法检测血清中IL-1β和IL-18的水平；空斑法测定感染小鼠唾液腺感染性病毒滴度.结果 MCMV感染后第3、5、7天,小鼠唾液腺组织中感染性病毒滴度逐渐增加；MCMV感染鼠脾脏巨噬细胞中AIM2、ASC和caspase-1蛋白的表达呈现一致的变化,与模拟感染对照鼠比较,AIM2、ASC和caspase-1蛋白相对吸光值在感染后第1天开始升高(P＞0.05),第3天明显升高并达峰值[分别为(1.121±0.243) vs(0.240±0.046),(1.318±0.333) vs (0.248±0.090),(1.085±0.243) vs (0.247±0.064); P＜0.01],其后接近正常；MCMV感染鼠血清IL-1β和IL-18水平在感染后第3天也明显高于模拟感染对照鼠[分别为(112.72±5.20) pg/ml vs (47.86±4.35) pg/ml,(42.74±4.23) pg/ml vs( 22.60±2.82)pg/ml；P＜0.01],其后均逐渐下降接近正常.结论 MCMC感染早期巨噬细胞通过AIM2炎性体识别胞浆MCMV DNA,可能成为CMV感染及感染后所引起的疾病的治疗靶点.
목적 관찰AIM2( absent in melanoma 2)재감염조기식별포장소서거세포병독(MCMV) DNA적변화상황.방법 건립MCMV전신파산형감염모형,접충MCMV Smith주후제1、3、5화7천각처사3지소서；동시설모의감염소서작위정상대조.Western blot검측비장거서세포중AIM2、함접자조망상관점양단백(ASC)화caspase-1단백적표체상황；동시응용ELISA법검측혈청중IL-1β화IL-18적수평；공반법측정감염소서타액선감염성병독적도.결과 MCMV감염후제3、5、7천,소서타액선조직중감염성병독적도축점증가；MCMV감염서비장거서세포중AIM2、ASC화caspase-1단백적표체정현일치적변화,여모의감염대조서비교,AIM2、ASC화caspase-1단백상대흡광치재감염후제1천개시승고(P＞0.05),제3천명현승고병체봉치[분별위(1.121±0.243) vs(0.240±0.046),(1.318±0.333) vs (0.248±0.090),(1.085±0.243) vs (0.247±0.064); P＜0.01],기후접근정상；MCMV감염서혈청IL-1β화IL-18수평재감염후제3천야명현고우모의감염대조서[분별위(112.72±5.20) pg/ml vs (47.86±4.35) pg/ml,(42.74±4.23) pg/ml vs( 22.60±2.82)pg/ml；P＜0.01],기후균축점하강접근정상.결론 MCMC감염조기거서세포통과AIM2염성체식별포장MCMV DNA,가능성위CMV감염급감염후소인기적질병적치료파점.
Objective To observe the changes of AIM2 ( absent in melanoma 2) inflammasome during early murine cytomegalovirus (MCMV) infection.Methods BALB/c mice were randomly divided into two groups.One was infected with MCMV Smith for establishing disseminated infection,the other was sham-inoculated control.On days 1,3,5 and 7 of the experiment,three mice of each group were randomly chosen to be killed separately.The expression of AIM2,ASC and caspase-1 in splenic macrophages was detected by Western blot,the levels of IL-1β and IL-18 in sera were measured by double antibody sandwich ELISA,and the viral titers in salivary gland tissues were quantified by a standard plaque assay.Results The MCMV titers in salivary gland tissues were gradually increased in MCMV-infected mice on days 3,5 and 7,while the expressions of AIM2 in macrophages were began to increase on day 1 and significantly increased and reached the highest level on day 3 but gradually decreased afterwards.The relative intensity of AIM2 on day 3 differed significantly between the MCMV-infected mice and the controls (1.121±0.243 vs 0.240±0.046,P＜0.01,t test),as did ASC ( 1.318±0.333 vs 0.248±0.090,P＜0.01 ) and caspase-1 ( 1.085±0.243 vs 0.247±0.064,P＜0.01 ).Meanwhile,the levels of IL-1β and IL-18 in MCMV-infected mice were (112.72±5.20) pg/ml and (42.74±4.23) pg/ml,and the levels were significantly higher (P＜0.01 ) than those in controls [ (47.86±4.35) pg/ml and (22.60±2.82) pg/ml].Conclusion These results demonstrate that AIM2 inflammasome is activated in macrophages during early MCMV infection and could be as a therapeutic target for CMV-induced diseases.