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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4 h-再灌注2 h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4 h再灌注2 h)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。
Objective To investigate the protective effect of hydroxysafflor yellow A on the injury of adrenal pheochromocytoma cells induced by oxygen-glucose deprivation and reperfusion and the effect of matrix metalloproteinase-9 (MMP-9). Methods Adrenal pheochromocytoma cells were subjected to hypoxia / glucose deprivation / reperfusion injury to simulate cerebral ischemia / reperfusion injury. Tetramethylbenzothiazolyl tetrazolium assay was used to detect adrenal pheochromocytoma cell survival rate. Lactate dehydrogenase Chromocytoma cell death rate, Hoechst33342 detection of adrenal pheochromocytoma cell apoptosis; immunocytochemistry of adrenal pheochromocytoma cells after induction of hypoxia-glucose deprivation reperfusion matrix metalloproteinase-9 expression. Results Oxygen-deprivation-glucose reperfusion induced the injury of adrenal pheochromocytoma cells. Compared with the normal control group, the cell survival rate was (75.5 ± 3.7)% at 4 h after reperfusion and was significantly decreased P <0.01). Tetramethythiazole blue method, lactate dehydrogenase method and Hoechst33342 staining results show that 1μmol·L -1 hydroxysafflor yellow A pretreatment can increase the survival of adrenal pheochromocytoma cells after induction of hypoxia and hypoglycemia reperfusion (90.8 ± 7.6)%, decreased the lactate dehydrogenase release rate (109.5 ± 7.0)% and decreased the apoptosis rate of adrenal pheochromocytoma cells (19.7 ± 7.5% h reperfusion 2 h), there was a significant difference (P <0.01). Matrix metalloproteinase-9 was induced by hypoxia-glucose deprivation (IR) -reperfusion, while hydroxysafflor yellow A inhibited its expression. CONCLUSION: Hydroxysafflor yellow A has a significant protective effect on the injury of adrenal pheochromocytoma cells induced by oxygen-glucose deprivation and glucose reperfusion, which can reduce the apoptosis of cells. The mechanism may be related to the inhibition of the expression of MMP-9.