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目的研究热射病小鼠脑组织中小胶质细胞活化的变化情况,初步探讨小胶质细胞在热射病脑损伤中的作用。方法采用小动物环境模拟舱建立温度(41.2±0.5)℃,相对湿度(60±2)%的热暴露损伤模型,将57只BALB/c雄性小鼠分为正常组和热射病1、6、24 h组。Real-time PCR检测大脑皮层M1型极化标志物TNF-α、CD45和CD11b的表达,以及M2型极化标志物Arg1、FIZZ和CD206的表达。Western blot检测各组大脑皮层CD45、CD11b、FIZZ和CD206的表达。免疫组织荧光技术检测CD45和CD206的分布和共定位表达。结果 Real-time PCR检测结果显示M1型极化标志物TNF-α、CD45和CD11b在热损伤后1 h达高峰,而M2型极化标志物Arg1、FIZZ和CD206在热损伤后24 h达高峰(P<0.01)。Western blot检测M1型极化标志物CD45及CD11b的蛋白表达量,在1 h后显著升高,随后逐渐下降,24 h后最低;M2型极化标志物FIZZ与CD206的表达趋势并不完全一致,但二者均在24 h后显著升高并达峰值(P<0.01)。免疫组织荧光染色结果显示CD45和CD206共定位表达于热射病小鼠大脑皮层,且分别在热损伤后1 h和24 h荧光光密度增强。结论热射病后小鼠脑组织小胶质细胞出现明显活化,其活化的规律表现为热射病后1 h主要为M1型极化,而在热射病后24 h则主要表现为M2型极化,这种变化形式可能与热射病后出现的中枢神经损伤有关。
Objective To study the changes of microglial activation in brain tissue of mice with thermo-rash disease and to investigate the role of microglia in brain injury induced by heat-jet. Methods The model of heat exposure was set up in a small animal environment simulation cabin at temperature (41.2 ± 0.5) ℃ and relative humidity (60 ± 2)%. Fifty-seven BALB / c male mice were divided into normal group and pyrogen- , 24 h group. Real-time PCR was used to detect the expression of M1-type polar markers of cerebral cortex, TNF-α, CD45 and CD11b, and the expression of M2-type polarized markers Arg1, FIZZ and CD206. Western blot was used to detect the expression of CD45, CD11b, FIZZ and CD206 in each group. Immunofluorescence was used to detect the distribution and co-localization of CD45 and CD206. Results Real-time PCR results showed that M1-type polarized markers TNF-α, CD45 and CD11b reached the peak at 1 h after heat injury, while M2-type polarized markers Arg1, FIZZ and CD206 peaked 24 h after heat injury (P <0.01). The protein expression levels of M1 polarization markers CD45 and CD11b were detected by Western blot and increased significantly after 1 h, then decreased gradually and reached the lowest after 24 h. The expression trend of M2-type polarization markers FIZZ and CD206 was not completely consistent , But both significantly increased and peaked at 24 h (P <0.01). Immunofluorescence staining showed that co-localization of CD45 and CD206 was detected in the cerebral cortex of pyramidal mice, and the optical density of fluorescence was enhanced at 1 h and 24 h after heat injury respectively. Conclusions The microglial cells in mouse brain showed obvious activation after heat stroke, and the activation pattern was mainly M1 polarization at 1 h after heat injection and M2 at 24 h after heat injection Polarization, this change may be associated with central nervous system damage after heat stroke.