Wu K Y, Ren Y X, Ruan Y M, Ma J L, Li C X, Wang Z
Institute of Occupational and Environmental Health, Twelfth People's Hospital Affiliated to Guangzhou Medical University, Guangzhou 510620, China.
Key Laboratory of Occupational Environment and Health, Guangzhou Twelfth People's Hospital, Guangzhou 510620, China.
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2024 May 20;42(5):332-339. doi: 10.3760/cma.j.cn121094-20230915-00157.
To explore the effect of the absent in melanoma 2 (AIM2) -mediated neuroinflammation in noise-induced cognitive dysfunction in rats. In April 2023, sixteen male Wistar rats were randomly divided into control group and noise group, with 8 rats in each group. The rats in the noise group were placed in 50 cm×50 cm×40 cm transparent boxes and exposed to 100 dB (A) white noise with a sound pressure level of 100 dB (A) (4 h/d for 30 d) . At the same time, rats in the control group were kept in similar boxes with environmental noise less than 60 dB (A) . After 30 days of noise exposure, the Morris water maze experiment was applied to test the learning and memory abilities of the rats; the pathological morphology of hippocampal tissues was observed by Hematoxylin-Eosin (HE) staining. Western blot was used to detect the protein expression levels of AIM2, cysteinyl aspartate specific proteinase-1 (caspase-1) , apoptosis-associated speck-like protein (ASC) , interleukin-1β (IL-1β) , IL-18, ionic calcium-binding articulation molecule-1 (Iba-1) , and glial fibrillary acidic protein (GFAP) . The expression of both Iba-1 and GFAP in hippocampal tissue was assessed by immunohistochemical staining. The co-localization of AIM2 with Iba-1 or GFAP was determined by immunofluorescence double staining. Compared with the control group, the escape latency of rats in the noise group was increased by 16.29 s, 17.71 s, and 20.26 s on days 3, 4, and 5, respectively. On day 6, the noise-exposed rats spent shorter time in the target quadrant and had fewer times in crossing the platform[ (7.25±2.27) s and (1.13±0.64) times] than the control group[ (15.64±3.99) s and (4.25±2.12) times] (<0.05) . After noise exposure, hippocampal neurons of rats displayed marked nuclear hyperchromatic and pyknosis phenomenon. The noise-exposed rats had higher numbers of both microglia and astrocytes (27.00±2.65 and 43.33±5.51) in the DG area of the hippocampus relative to the control group (14.67±3.06 and 20.00±4.58) (<0.05) . Moreover, the glial cells in the noise group had larger cell cytosol with more and thicker branches. The protein expression levels of inflammatory cytokines Cleaved-IL-1β and Cleaved-IL-18 in the hippocampus of rats in the noise group (1.55±0.19 and 1.74±0.12) were significantly higher than the control group (1.00±0.11 and 1.00±0.13) (<0.05) . After noise exposure, the protein expression levels of AIM2, Cleaved-Caspase-1 and ASC (1.19±0.09, 1.34±0.07 and 1.14±0.01) were higher than the control group (1.00±0.07, 1.00±0.14 and 1.00±0.06) and differences between the two groups were statistically significant (<0.05) . A significant increase in the number of cells co-localizing AIM2 with Iba-1 or GFAP in the noise group (28.67±4.04 and 40.67±5.13) compared with the control group (15.67±4.04 and 17.67±3.79) , and statistically significant differences were observed between the two groups (<0.05) . Noise exposure may activate the AIM2 inflammasome in hippocampal glial cells of rats, releasing excessive inflammatory cytokines and causing neuroinflammation that damages neurons.
探讨黑色素瘤缺失2(AIM2)介导的神经炎症在大鼠噪声性认知功能障碍中的作用。2023年4月,将16只雄性Wistar大鼠随机分为对照组和噪声组,每组8只。将噪声组大鼠置于50 cm×50 cm×40 cm的透明箱中,暴露于声压级为100 dB(A)的白噪声(100 dB(A),4 h/d,共30 d)。同时,将对照组大鼠置于环境噪声小于60 dB(A)的类似箱中。噪声暴露30天后,应用Morris水迷宫实验测试大鼠的学习和记忆能力;采用苏木精-伊红(HE)染色观察海马组织的病理形态。采用蛋白质印迹法检测AIM2、半胱天冬酶-1(caspase-1)、凋亡相关斑点样蛋白(ASC)、白细胞介素-1β(IL-1β)、IL-18、离子钙结合衔接分子-1(Iba-1)和胶质纤维酸性蛋白(GFAP)的蛋白表达水平。通过免疫组织化学染色评估海马组织中Iba-1和GFAP的表达。通过免疫荧光双染色确定AIM2与Iba-1或GFAP的共定位。与对照组相比,噪声组大鼠在第3、4和5天的逃避潜伏期分别增加了16.29 s、17.71 s和20.26 s。在第6天,噪声暴露大鼠在目标象限的停留时间较短,穿越平台的次数较少[(7.25±2.27)s和(1.13±0.64)次],而对照组为[(15.64±3.99)s和(4.25±2.12)次](<0.05)。噪声暴露后,大鼠海马神经元出现明显的核染色质增多和固缩现象。与对照组(14.67±3.06和20.00±4.58)相比,噪声暴露大鼠海马齿状回(DG)区的小胶质细胞和星形胶质细胞数量均较高(27.00±2.65和43.33±5.51)(<0.05)。此外,噪声组的神经胶质细胞胞质较大,分支更多更粗。噪声组大鼠海马中炎性细胞因子裂解型IL-1β和裂解型IL-18的蛋白表达水平(1.55±0.19和1.74±0.12)显著高于对照组(1.00±0.11和1.00±0.13)(<0.05)。噪声暴露后,AIM2、裂解型Caspase-1和ASC的蛋白表达水平(1.19±0.09、1.34±0.07和1.14±0.01)高于对照组(1.00±0.07、1.00±0.14和1.00±0.06),两组间差异具有统计学意义(<0.05)。与对照组(15.67±4.04和17.67±3.79)相比,噪声组中AIM2与Iba-1或GFAP共定位的细胞数量显著增加(28.67±4.04和40.67±5.13),两组间差异具有统计学意义(<0.05)。噪声暴露可能激活大鼠海马神经胶质细胞中的AIM2炎性小体,释放过多炎性细胞因子并引起神经炎症,从而损害神经元。