Circhipk3对热射病神经损伤中小胶质细胞极化的影响

目的探讨circhipk3在热射病神经损伤中小胶质细胞的表达情况,初步分析circhipk3对热射病神经损伤中小胶质细胞极化的影响。方法随机(随机数字法)将小鼠分为对照组及热射病0.8 h组(HS 0.8),热射病8 h组(HS 8),热射病24 h组(HS 24)。通过建立小鼠热射病(HS)模型,取得热损伤脑组织,分离小胶质细胞并提取RNA。定量PCR法检测小胶质细胞中M1和M2标志分子,评估小胶质细胞极化方向及类型。检测circhipk3在热射病神经损伤中小胶质细胞的表达水平,通过干预circhipk3在小胶质细胞中的表达,进一步阐明circhipk3对小胶质细胞极化的影响。结果HS 8组在CD45、CD11-b表达较正常组明显上升(4.41±0.18)vs.(1±0.15),P=0.000]、(3.47±0.19)vs.(1±0.15),P=0.000],而HS 24组的CD45、CD11-b较HS 8组明显下降(1.34±0.15)vs.(4.41±0.18),P=0.000]、(1.38±0.21)vs.(3.47±0.19),P=0.001]。同时HS 8组在CD206、FIZZ、Arg1表达较对照组开始上升(1.59±0.16)vs.(1±0.12),P=0.014]、(1.62±0.15)vs.(1±0.15),P=0.002]、(2.23±0.28)vs.(1±0.19),P=0.004],而HS 24组的CD206、FIZZ、Arg1较对照组显著升高(2.67±0.20)vs.(1±0.12),P=0.002]、(2.19±0.15)vs.(1±0.15),P=0.000]、(3.04±0.18)vs.(1±0.19),P=0.001];circhipk3 mimicis显著增高Arg1表达(7.26±0.06)vs.(3.86±0.06),P=0.000];同时circhipk3 inhibitor促进CD45及HO-1表达(2.96±0.03)vs.(1.63±0.09),P=0.000]、(2.52±0.10)vs.(1.30±0.02),P=0.000]。结论热射病早期神经损伤中小胶质细胞以M1型为主,HO-1可能是小胶质细胞M1型标记物之一,circhipk3在小胶质细胞中高表达主要促进其向M2型转化。

Effect of circhipk3 on polarization of microglial cells in nerve injury caused by heat radiation

Objective:To investigate the expression of circhipk3 in microglial cells in heat-induced neurological injury, and to preliminary analyze the effect of circhipk3 on microglial polarization in heat-induced neurological injury.Methods:Mice were randomly (random number) divided into a control group and a heat radiation disease 0.8 h group (HS 0.8), a heat radiation disease 8h group (HS 8), and a heat radiation disease 24 h group (HS 24). By establishing a mouse model of heat shock (HS), heat-damaged brain tissue was obtained, microglia were isolated and RNA was extracted. Quantitative PCR method was used to detect M1 and M2 marker molecules in microglia, and to evaluate the polarization direction and type of microglia. The expression level of circhipk3 was detected in microglial cells in heat-induced neurological injury, and the effect of circhipk3 on microglial polarization was further elucidated by intervening the expression of circhipk3 in microglial cells.Results:The expression of CD45 and CD11-b in the HS 8 group was significantly higher than that in the control group (4.41±0.18) vs. (1±0.15), P=0.000], (3.47±0.19) vs (1±0.15), P=0.000] , and the CD45 and CD11-b of the HS 24 group was significantly lower than that of the HS 8 group (1.34±0.15) vs. (4.41±0.18), P=0.000], (1.38±0.21) vs. (3.47±0.19), P= 0.001]. At the same time, the expression of CD206, FIZZ and Arg1 in the HS 8 group started to increase compared with the control group (1.59±0.16) vs. (1±0.12), P=0.014], (1.62±0.15) vs. (1±0.15), P=0.002 ], (2.23±0.28) vs. (1±0.19), P=0.004], and CD206, FIZZ, and Arg1 in the HS 24 group were significantly higher than those in the control group (2.67±0.20) vs. (1±0.12), P=0.002], (2.19±0.15) vs. (1±0.15), P=0.000], (3.04±0.18) vs. (1±0.19), P=0.001]; circhipk3 mimicis significantly increased the expression of Arg1 (7.26± 0.06) vs. (3.86±0.06), P=0.000]; at the same time, circhipk3 inhibitor promoted the expression of CD45 and HO-1 (2.96±0.03) vs. (1.63±0.09), P=0.000], (2.52±0.10) vs. ( 1.30±0.02), P=0.000]. Conclusions:Microglial cells are predominantly M1-type in early neurological injury of heat radiation disease. HO-1 may be one of the microglial M1-type markers. The high expression of circhipk3 in microglial cells mainly promotes its transformation to M2 type.