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目的研究嘌呤受体P2x7在小胶质细胞炎症反应中的作用。方法通过分子克隆的方法构建pCDNA3.1-P2x7重组质粒,构建成功后经测序确认质粒序列无误,通过Lipo2000转染小胶质细胞株BV-2细胞,采用PCR方法测定BV-2细胞内P2x7的表达,Westernblot法测定P2x7和Cox-2的蛋白表达。结果细胞经重组质粒转染成功后,pCDNA3.1-P2x7组P2x7和Cox-2蛋白表达水平分别为(4.21±0.13)%和(2.60±0.05)%,高于pCDNA3.1组的(2.61±0.05)%和(1.62±0.04)%,差异均有统计学意义(P<0.05)。结论通过过表达P2x7受体,能引起小胶质细胞的激活,提示可进一步引发炎症反应。
Objective To investigate the role of purine receptor P2x7 in the microglial inflammatory response. Methods The recombinant plasmid pCDNA3.1-P2x7 was constructed by molecular cloning. After successful construction, the plasmid sequence was confirmed by sequencing. The microglia cell line BV-2 was transfected with Lipo2000, and the expression of P2x7 in BV-2 cells The protein expression of P2x7 and Cox-2 was detected by Western blot. Results After transfected with recombinant plasmid, the expression levels of P2x7 and Cox-2 in pCDNA3.1-P2x7 group were (4.21 ± 0.13)% and (2.60 ± 0.05)%, respectively, higher than that in pCDNA3.1 group (2.61 ± 0.05)% and (1.62 ± 0.04)% respectively, the difference was statistically significant (P <0.05). Conclusion Overexpression of P2x7 receptor can cause microglial activation, suggesting further inflammatory response.