黄芩苷对白色念珠菌酶活性的影响

Effect of Baicalin on Enzyme Activity of Candida albicans

目的运用分光光度法检测黄芩苷作用白色念珠菌后,其琥珀酸脱氢酶(SDH)、细胞色素C氧化酶(CCO)、Ca2+-Mg2+-ATPase活性的改变,以探讨黄芩苷抗白色念珠菌作用靶位,为探索中药活性成分抗真菌作用机理提供依据。方法收集经黄芩苷作用了相应时间的白色念珠菌,制备菌悬液,反复冻融研磨,差速离心法提取白色念珠菌线粒体;Lowry法检测线粒体蛋白质含量;分光光度法检测SDH酶、CCO酶、Ca2+-Mg2+-ATPase活力。结果①0.25,0.5,1mg/ml黄芩苷浓度组白色念珠菌SDH活力明显低于0mg/ml黄芩苷浓度组(P<0.05),且出现黄芩苷浓度越高SDH活力越低的趋势。不同作用时间组间白色念珠菌SDH活力差异无统计学意义(P>0.05)。②白色念珠菌经不同浓度黄芩苷作用不同时间后,其CCO酶活力均无明显差异(P>0.05)。③0.25,0.5,1mg/ml黄芩苷浓度组与0mg/ml黄芩苷浓度组比较,Ca2+-Mg2+-ATPase活力差异显著,黄芩苷浓度越高,Ca2+-Mg2+-ATPase活力越低,作用时间对Ca2+-Mg2+-AT-Pase活力没有明显影响(P>0.05)。结论①黄芩苷具有明显的抗白色念珠菌作用;②黄芩苷可降低白色念珠菌SDH酶活力;③黄芩苷对白色念珠菌CCO酶活力未见明显影响;④黄芩苷可降低白色念珠菌Ca2+-Mg2+-ATPase活力。

Objective To investigate the target site of baicalin against Candida albicans and supply base for exploring antifungal action mechanism of the traditional Chinese medicine active component by detecting the effect of baicalin on succinate dehydrogenase, cytochrome - oxydase, Ca^2＋-Mg^2＋-ATPase through absorption spectrometry. Methods After incubated with different concentrations of baicalin for corresponding time, Candida albicans was harvested and resuspended in physiological saline,which was followed by freeze thawing and grind three times. Then mitochordria was extracted by different centrifugation and used as sample. Lowry method was applied to detect the protein content and absorption spectrometry was used to detect the activities of SDH, CCO, Ca^2＋-Mg^2＋-ATPase. Results It was showed that （1） the activity of SDH of Candida albicans in control group was much higher than that of Candida albicans treated with baicalin at three doses of 0.25 mg/ml,0.5mg/ml, 1 mg/ml,while no difference was observed among the three doses. （2）No difference was observed in the activity of CCO of Candida albicans treated with different doses of baicalin（ P 〉 0.05 ） for different action time. （3）The activity of Ca^2＋-Mg^2＋-ATPase decreased significantly in baicalin groups while compared with that in control group, the higher the concentration of baicalin was , the lower the activity of Ca^2＋-Mg^2＋-ATPase was. Conclusion （1） Baicalin has significant antifungal effect on Candida albicans;（2） baicalin can decrease the activity of SDH of Candida albicans ;（3） baicalin has no effect on the activity of CCO of Candida albicans ;（4） baicalin can decrease the activity of Ca^2＋-Mg^2＋-ATPase of Candida albicans.