PI3K/Akt通路介导LPS调节星形胶质细胞HSPA12B表达的研究

Effect of PI3K/Akt pathway on LPS-induced HSPA12B expression in rat astrocytes

目的探讨磷脂酰肌醇-3-激酶/丝氨酸-苏氨酸蛋白kinase B(PI3K/Akt)通路对内毒素脂多糖(LPS)诱导的星形胶质细胞热休克蛋白A12B(HSPA12B)表达的影响。方法将处于对数生长期的新生SD大鼠星形胶质细胞分为3组:con组(空白对照)、LPS组(1μg·ml^(-1)LPS刺激4h)、LPS+LY组(20μmol·L^(-1)LY294002预处理1h+1μg·ml^(-1)LPS刺激4h)。采用Western blot法检测3组细胞HSPA12B、磷酸化Akt(p-Akt)蛋白表达水平,采用细胞免疫荧光染色法检测细胞HSPA12B免疫荧光强度,并进行比较。结果 3组星形胶质细胞HSPA12B、p-Akt蛋白表达水平比较差异均有统计学意义(均P<0.05);与con组比较,LPS组星形胶质细胞HSPA12B、p-Akt蛋白表达水平均上调(均P<0.05);而与LPS组比较,LPS+LY组星形胶质细胞HSPA12B、p-Akt蛋白表达水平均下调(均P<0.05)。LPS组星形胶质细胞HSPA12B荧光强度强于con组,而LPS+LY组星形胶质细胞HSPA12B荧光强度较LPS组减弱。结论 LPS或是通过PI3K/Akt通路介导调节星形胶质细胞HSPA12B的表达,进而影响中枢神经系统炎症过程。

Objective To investigate the effect of phosphatidylinositol 3-kinase/serine threonine protein kinaseB（PI3K/Akt） on the expression of Heat shock protein A 12B （HSPA12B） in rat astrocytes induced by lipopolysaccharide （LPS）.Methods Astrocytes in logarithmic growth phase from neonatal SD rats were divided into 3 groups： control group, LPS group（1μg· ml^-1 LPS-stimulated for 4 h） and LPS＋LY group （20μmol·L^-1 LY2940021μg·ml^-1 pretreated for 1 h ＋ LPS-stimulated for4h）. The expression of HSPA12B and phosphorylated Akt （p-Akt） protein was detected by Western blot and the intensity change ofHSPA12B was detected by cell immunofluorescence staining. Results There were significant differences in the expression ofHSPA12B, p-Akt protein in astrocytes among three groups （all P〈0.05）. Compared with control group, the expression ofHSPA12B, p-Akt protein in astrocytes was increased in LPS group （all P〈0.05）; compared with LPS group, the expression ofHSPA12B, p-Akt protein was reduced in LPS＋LY group（P〈0.05）. The fluorescence intensity of HSPA12B protein in LPS groupwas stronger than that in control group, while the HSPA12B intensity in LPS＋LY group was lower than that in LPS group. Conclu-sion PI3K/Akt pathway may be involved in LPS-induced HSPA12B expression in astrocytes, which may affect the inflammationprocess of central nervous system.