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目的考察缝隙连接蛋白43(connexin43,Cx43)基因敲除对脂多糖(lipopolysacchiride,LPS)腹腔注射后胶质细胞的活化和脑内炎症反应的影响。方法采用腹腔LPS注射方法制作外周免疫激活小胶质细胞模型,实验动物共分为4组。Cx43基因敲除小鼠LPS腹腔注射组(Cx43KO-LPS组),Cx43基因敲除小鼠生理盐水对照组(Cx43 KO-NS组),野生小鼠LPS腹腔注射组(WT-LPS组)和野生小鼠生理盐水对照组(WT-NS组)。通过免疫荧光染色的方法观察小胶质细胞(Iba1)和星形胶质细胞(GFAP),通过Western blot方法考察脑内炎症介质IL-1β的变化。结果免疫荧光染色结果提示,Iba1和GFAP的荧光光密度在Cx43 KO-LPS组显著高于Cx43 KO-NS组,WT-LPS组显著高于WT-NS组,WT-LPS组显著高于Cx43KO-LPS组。IL-lβ水平在腹腔注射LPS的2组均高于2个生理盐水对照组,但在WT-LPS组具有最高的表达。结论 Cx43基因敲除能够减轻LPS腹腔注射后脑内炎症介质IL-1β的表达升高,减轻小胶质细胞和星形胶质细胞的活化。
Objective To investigate the effect of connexin43 (Cx43) knockout on activation of glial cells and inflammatory reaction in the brain after intraperitoneal injection of lipopolysacchride (LPS). Methods Peripheral immune activation microglial cells model was made by intraperitoneal injection of LPS. Experimental animals were divided into 4 groups. Cx43 knockout mice were injected intraperitoneally with LPS (Cx43KO-LPS), Cx43 knockout mice (Cx43 KO-NS), wild type LPS (WT-LPS) and wild Mouse saline control group (WT-NS group). Immunofluorescence staining was used to observe the changes of intracellular inflammatory mediators IL-1β by immunofluorescence staining of microglial cells (Iba1) and astrocytes (GFAP). Results The results of immunofluorescence staining showed that the fluorescence intensity of Iba1 and GFAP in Cx43 KO-LPS group was significantly higher than that in Cx43 KO-NS group, the WT-LPS group was significantly higher than that in WT-NS group, and the WT-LPS group was significantly higher than Cx43KO- LPS group. The level of IL-1β in the LPS group was higher than that in the two normal saline control groups, but highest in the WT-LPS group. Conclusion Cx43 gene knockdown can reduce the expression of IL-1β in the intracerebral inflammatory mediators after intraperitoneal injection of LPS, and reduce the activation of microglia and astrocytes.