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目的了解β_2肾上腺素能受体激动剂特布他林对烧伤血清和去甲肾上腺素(NE)诱导神经胶质细胞血管内皮生长因子(VEGF)基因表达的影响。方法制备烧伤大鼠及正常大鼠血清。取出生1～3 d乳鼠脑组织行原代神经胶质细胞培养,并在培养液中加入不同的刺激物,分为:(1)对照组:加入体积分数10%的正常大鼠血清;(2)NE1、NE2、NE3组:分别加入体积分数10%烧伤血清+10、20、50μmol/L NE;(3)TBN1、TBN2、TBN3组:分别加入10、20、50μmol/L特布他林+体积分数10%烧伤血清+10、20、50μmol/L NE。采用蛋白质印迹法检测各组VEGF蛋白的表达;实时荧光定量PCR法检测VEGF mRNA的表达。结果对照组VEGF蛋白的表达处于较低水平,NE1、NE2、NE3组其表达随NE浓度的增大逐渐增加,而TBN1、TBN2、TBN3组的表达明显增加。对照组神经胶质细胞VEGF mRNA表达量[(5.72±0.12)拷贝数/g]较低,NE1、NE2、NE3组VEGF mRNA的表达[(13.26±0.03)、(10.37±0.04)、(14.87±0.55)拷贝数/g]均高于对照组(P<0.01)。而TBN1、TBN2、TBN3组VEGF mRNA表达量[(13.39±0.19)、(15.77±0.11)、(16.00±0.07)拷贝数/g]也逐渐增加,除TBN1组外其余2组与各NE组比较,差异有统计学意义(P<0.01)。结论特布他林可增强烧伤血清和NE诱导神经胶质细胞VEGF基因的表达。
Objective To investigate the effects of β 2 adrenergic receptor agonist terbutaline on the gene expression of vascular endothelial growth factor (VEGF) induced by burn serum and norepinephrine (NE) in glial cells. Methods Burn rats and normal rat serum were prepared. Primary brain cells of neonatal rats were sacrificed 1 ~ 3 days after primary glial cell culture, and different stimuli were added to the culture medium, divided into: (1) control group: the volume of 10% normal rat serum was added; (2) NE1, NE2 and NE3 groups: 10%, 50%, and 50% NE respectively; (3) TBN1, TBN2 and TBN3 groups were treated with 10, 20 and 50μmol / Lin + volume fraction 10% burn serum + 10, 20, 50 μmol / L NE. The expression of VEGF protein in each group was detected by Western blotting. The expression of VEGF mRNA was detected by real-time fluorescence quantitative PCR. Results The expression of VEGF protein in the control group was at a low level. The expression of VEGF protein in NE1, NE2 and NE3 groups increased gradually with the increase of NE concentration, but significantly increased in TBN1, TBN2 and TBN3 groups. The expression of VEGF mRNA in glial cells in control group [(5.72 ± 0.12) copies / g] was lower than that in NE1, NE2 and NE3 groups [(13.26 ± 0.03), (10.37 ± 0.04), (14.87 ± 0.55 copies / g] were higher than the control group (P <0.01). The expression of VEGF mRNA in TBN1, TBN2 and TBN3 groups was also increased ([(13.39 ± 0.19), (15.77 ± 0.11) and (16.00 ± 0.07) copies / g] , The difference was statistically significant (P <0.01). Conclusion Terbutaline can enhance the expression of VEGF gene induced by burn serum and NE in glial cells.