蒙药诃子汤制草乌总生物碱对乳大鼠心肌细胞的毒性作用研究

目的 探讨蒙药诃子汤炮制草乌总生物碱对心肌细胞的毒性作用.方法 制备乳大鼠原代心肌细胞,胎盘蓝染色法检测心肌细胞存活率,免疫组织化学染色法鉴定心肌细胞.制备的原代细胞培养3-4d,分别加入诃子汤制草乌、生草乌总生物碱0.5、0.25、0.125、0.0625 mg/mL培养30、60 min,MTT法检测心肌细胞存活率,比色法检测乳酸脱氢酶(LDH)漏出率,Hoechst33528荧光染色法检测细胞核凋亡率.结果 诃子汤制草乌总生物碱、生草乌总生物碱在与心肌细胞共同孵育30 min、60 min时均能降低心肌细胞活力,细胞LDH漏出率增加,发生细胞凋亡；诃子汤制草乌总生物碱所致的心肌细胞毒性作用弱于生草乌总生物碱.结论 经诃子汤炮制的草乌总生物碱对心肌细胞的毒性作用明显低于生草乌总生物碱,表明采用诃子汤炮制确能降低草乌的心肌毒性.

Toxicity of total alkaloids of Aconitum kusnezoffii Reichb processed with myrobalan on primary cultured myocardial cells of neonatal rats

AIM To evaluate the toxicity of total alkaloids of Aconitum kusnezoffii Reichb processed with myrobalan(TAAM) to myocardial cells in vitro.METHODS The primary myocardial cells of neonatal rats were carried out.The cell survival rate was detected using the trypan blue staining method,and myocardial cells were identified by anti-α-sarcomeric actin and anti-fibronectin antibodies with immunochemical staining assay.When cells were cultured for 3-4 d,0.5 mg/mL,0.25 mg/mL,0.125 mg/mL and 0.062 5 mg/mL of total alkaloids of TAAM,total alkaloids of Aconitum kusnezoffii Reichb(TAA) were incubated with myocardial cells,respectively.After the cells were cultured with TAAM or TAA for 30,60 min,the cell viability was assayed with MTT,and the release rate of lactate dehydrogenase(LDH) was assayed with biochemical method.The apoptosis rate was observed with Hoechst33528.RESULTS TAAM and TAA could decrease the cell viability,and increase the release rate of LDH in 30,60 min,and also lead to apoptosis.But the toxicity of TAAM to myocardial cells was less than that of TAA.CONCLUSION Myrobalan can decrease the toxicity to myocardial cells.