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研究超声激活血卟啉对小鼠hepatoma22(H-22)肿瘤细胞抗氧化酶活性的影响,初步探讨其杀伤细胞的生物学机制。采用频率为1.43MHz、强度为2.0W/cm2的聚焦超声,结合100μg/ml的血卟啉作用于肿瘤细胞,通过台盼蓝拒染法检测细胞存活率;2′,7′-二氯荧光黄双乙酸盐(Dichlorofluorescein diacetata,DCFH-DA)活性氧检测试剂盒检测细胞内活性氧水平;酶化学方法检测细胞内几种抗氧化酶活性的变化。发现同等处理条件下,超声结合血卟啉对细胞的损伤显著高于单纯超声组,单纯血卟啉无明显的细胞毒作用;超声激活血卟啉后细胞悬液中活性氧的生成量明显升高,而细胞内超氧化物歧化酶(Superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(Glutathi-one peroxidase,GSH-PX)、过氧化氢酶(Catalase,CAT)活性均显著下降。推测超声激活血卟啉对小鼠H-22腹水瘤细胞的杀伤作用可能与肿瘤细胞中抗氧化物酶活性降低有关。
To investigate the effect of hematoporphyrin on the anti-oxidative enzyme activity of murine hepatoma22 (H-22) tumor cells and to explore the biological mechanism of hematopoietic cell killing. Focused ultrasound with a frequency of 1.43 MHz and an intensity of 2.0 W / cm 2 was used in tumor cells in combination with 100 μg / ml hematoporphyrin, and the cell viability was detected by trypan blue exclusion. The 2 ’, 7’- Dichlorofluorescein diacetata (DCFH-DA) active oxygen detection kit was used to detect intracellular reactive oxygen species (ROS) levels. Enzyme-linked immunosorbent assay was used to detect the changes of several antioxidant enzymes in cells. It was found that under the same treatment conditions, the damage of cells by ultrasound combined with hematoporphyrin was significantly higher than that of pure ultrasound group and pure hematoporphyrin. No significant cytotoxic effect was found when hematoporphyrin was activated by ultrasound. While the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-PX) and catalase (CAT) in cells were significantly decreased. It is speculated that the killing effect of ultrasound-activated hematoporphyrin on mouse H-22 ascites tumor cells may be related to the decrease of antioxidant enzyme activity in tumor cells.