PAR2在损伤的大鼠脊髓星形胶质细胞增生中的作用研究

Role of PAR2 in proliferation of injured spinal cord astrocytes of rats

摘要:

目的：探讨蛋白酶激活受体2（protease activated receptor 2，PAR2）对损伤的大鼠脊髓星形胶质细胞增生的作用。方法：原代培养得到高纯度（>98%）大鼠脊髓星形胶质细胞，建立体外星形胶质细胞损伤模型。星形胶质细胞损伤后立即给予生理盐水（损伤模型组）、PAR2特异性抑制剂FSLLRY-NH2（FSLLRY-NH2组）、PAR2特异性抗体Anti-PAR2 antibody（Anti-PAR2 an－tibody组）处理；对照组为原代培养的大鼠脊髓星形胶质细胞，不作任何处理。应用蛋白质印迹法及免疫荧光标记技术检测星形胶质细胞标志蛋白胶质纤维酸性蛋白（glial fibrillary acidic protein，GFAP）和波形蛋白（Vimentin）的表达情况，应用CCK-8法检测星形胶质细胞增殖活性。结果：损伤后24 h及72 h 对照组、FSLLRY-NH2组和Anti-PAR2 antibody组GFAP和Vimentin的表达均明显低于损伤模型组，损伤后12、24、72 h对照组、FSLLRY-NH2组和Anti-PAR2 antibody组星形胶质细胞增殖活性均明显低于损伤模型组（P<0.05）。结论：大鼠脊髓星形胶质细胞损伤后阻断PAR2可抑制其细胞增生。

Abstract:

Objective：To explore the role of protease activated receptor2（PAR2） in proliferation of injured spinal cord astrocytes of rats. Methods：High purity spinal cord astrocytes of rats were achieved from primary culture（purity>98%） and the injured astrocytes model in vitro was established. Injured astrocytes were treated by normal saline（injured group），PAR2 inhibitor FSLLRY-NH2（FS－LLRY-NH2 group） and PAR2 antibody Anti-PAR2 antibody（Anti-PAR2 antibody group） immediately. Primary culture cells without any treatment were included in control group. The expression levels of GFAP and Vimentin，marker proteins of astrocytes，were detected using the immunofluorescence staining and Western blot after injured astrocytes model being established and treated. Proliferative ac－tivity of astrocytes was detected by CCK-8 after injured astrocytes model being established and treated. Results：At 24 h and 72 h after injured astrocytes model being established，the expression levels of GFAP and Vimentin were significantly higher in injured group compared with those in control group，FSLLRY-NH2 group and Anti-PAR2 antibody group；proliferative activity of astrocytes was also significantly higher in injured group compared with that in control group，FSLLRY-NH2 group and Anti-PAR2 antibody group at 12，24，72 h after injured astrocytes model being established（P<0.05）. Conclusion：Blocking PAR2 can inhibit proliferation of injured spinal cord astrocytes of rats.