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目的观察超声激活血卟啉对新生SD大鼠心肌细胞的凋亡效应,并初步探讨其相关机制。方法采用差速贴壁法培养原代新生SD大鼠心肌细胞,将其分为4组:超声(频率300 kHz,声强1.5 W/cm2,辐照60 s)+血卟啉组(浓度20μg/m l),超声组(频率300 kHz,声强1.5 W/cm2,辐照60 s),血卟啉组(浓度20μg/m l)和对照组。采用MTT法观察各处理因素对细胞的生长抑制情况;流式细胞仪膜连蛋白V-FITC/PI双染法检测细胞凋亡状况;免疫荧光细胞化学法鉴定细胞及检测Fas/FasL蛋白表达情况。结果 MTT法测得24 h抑制率超声+血卟啉组显著高于其他各组(P<0.05)。膜连蛋白V-FITC/PI双染法检测超声+血卟啉组细胞早期凋亡率明显高于其他各组(P<0.05)。与其他各组相比,免疫荧光细胞化学法测定超声+血卟啉组Fas/FasL蛋白表达光密度值有明显增高(P<0.05)。结论超声激活血卟啉对心肌细胞有明显的凋亡效应,其可能与Fas/FasL蛋白表达增高有关。
Objective To observe the effect of hematoporphyrin activated by ultrasound on cardiomyocyte apoptosis in neonatal SD rats and to explore its underlying mechanism. Methods Cardiac myocytes of primary neonatal SD rats were cultured by differential adherence method and divided into 4 groups: ultrasound (frequency 300 kHz, intensity 1.5 W / cm2, irradiation 60 s) + hematoporphyrin group (concentration 20 μg / ml), ultrasound group (frequency 300 kHz, sound intensity 1.5 W / cm2, irradiation 60 s), hematoporphyrin group (20 μg / ml) and control group. MTT assay was used to observe the inhibition of cell growth by flow cytometry. Flow cytometry was used to detect apoptosis status by FITC / PI double staining. Immunofluorescence staining was used to identify cells and Fas / FasL protein expression . Results MTT assay 24 h inhibition rate ultrasound + hematoporphyrin group was significantly higher than the other groups (P <0.05). Annexin V-FITC / PI double staining detected ultrasound + hematoporphyrin group cells early apoptosis rate was significantly higher than the other groups (P <0.05). Compared with other groups, the expression of Fas / FasL protein in the ultrasound + hematoporphyrin group was significantly increased (P <0.05) by immunofluorescence cytochemistry. Conclusion Ultrasound-activated hematoporphyrin has obvious apoptosis effect on cardiomyocytes, which may be related to the increase of Fas / FasL protein expression.