目的探讨羟基红花黄色素A对缺氧缺糖再灌注诱导肾上腺嗜铬细胞瘤细胞损伤的保护作用及基质金属蛋白酶-9表达的影响。方法以肾上腺嗜铬细胞瘤细胞缺氧缺糖再灌注损伤模拟脑缺血再灌注损伤，四甲基偶氮唑蓝法检测肾上腺嗜铬细胞瘤细胞的存活率，乳酸脱氢酶法检测肾上腺嗜铬细胞瘤细胞死亡率，Hoechst33342检测肾上腺嗜铬细胞瘤细胞凋亡；细胞免疫化学法检测肾上腺嗜铬细胞瘤细胞经缺氧缺糖再灌注诱导后基质金属蛋白酶-9表达。结果缺氧缺糖再灌注可诱导肾上腺嗜铬细胞瘤细胞损伤，与正常对照组相比，缺氧缺糖4h-再灌注2h组细胞存活率为（75.5±3.7）%，明显降低（P<0.01）。四甲基偶氮唑蓝法、乳酸脱氢酶法及Hoechst33342染色结果显示，1μmol·L-1羟基红花黄色素A预处理能增加缺氧缺糖再灌注诱导后肾上腺嗜铬细胞瘤细胞存活率（90.8±7.6）%，降低细胞乳酸脱氢酶释放率（109.5±7.0）%，同时减少肾上腺嗜铬细胞瘤细胞凋亡率（19.7±7.5）%，与模型组（缺氧缺糖4h再灌注2h）相比，均具有显著性差异（P<0.01）。基质金属蛋白酶-9经缺氧缺糖再灌注处理后能产生诱导表达，而羟基红花黄色素A能抑制其表达。结论羟基红花黄色素A对缺氧缺糖再灌注诱导的肾上腺嗜铬细胞瘤细胞损伤具有明显保护作用，能减少细胞凋亡，其机制可能与抑制基质金属蛋白酶-9的诱导表达有关。
Abstract
OBJECTIVE To investigate the neuroprotective effects of hydroxysafflor yellow A(HSYA) on the injury of PC12 cells induced by oxygen-glucose deprivation and reperfusion, and the effect on the expression of metrix metalloproteinase-9(MMP-9) in PC12 cells. METHODS The PC12 cells were exposed to oxygen-glucose deprivation(OGD), and recoveried for different times. The PC12 cell viability and lactate dehydrogenase(LDH) release were seperately detected by MTT assays and LDH detecting kits, the apoptotic cells were double stained by propidumiodid(PI) and Hoechst33342, and the expression of MMP-9 was detected by immunocytochemistry. RESULTS Compared to the control group, in OGD 4 h-R 2 h group, cell viability significantly decreased to ,(P<0.01). Whereas HSYA improved the injuries induced by OGD-R, reflected on the obvious increased cell viability and reduced releasing of LDH (P<0.01) at the concentration of 1 μmol·L-1. And the result stained by PI and Hoechst33342 showed that HSYA also markedly inhibited the apoptosis of PC12 cell induced by OGD-R, which decreased to(19.7±7.5)% in HSYA 1 μmol·L-1 group(P<0.01). Meanwhile, the expression of active MMP-9 induced by OGD-R was decreased after the treatment of HSYA, similar to positive control edaravone. CONCLUSION It is proved that HSYA protects the injury and inhibits the apoptosis induced by OGD-R in PC12 cells, and the mechanism may be related to the inhibition of active MMP-9 expression.
关键词
羟基红花黄色素A /
缺血再灌注损伤 /
细胞凋亡 /
基质金属蛋白酶-9
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Key words
hydroxysafflor yellow A /
ischemia-reperfusion injury /
apoptosis /
metrix metalloproteinase-9
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中图分类号：
10 11669/cpj 2013 09 008
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参考文献
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脚注
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基金
湖州市自然科学基金资助项目（2009YZ11）；浙江省中医药科学研究基金（2011ZA101）
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