The Effect Of Indomethacin On The Hypha Growth Of Candida Albicans In Animal Experiment

The experiment is to observe the effect of indomethacin on the hypha growth of candida albicans in the estrogen-dependent murine model of candida albicans vaginitis, so as to explore the new therapy drug for candida albicans vaginitis, accordingly providing the new choose for the treatment of the monilial vaginitis in clinic. The experiment is divided by three parts: (1)Choosing the culture conditions of powerful inducing the hypha growth and anoxic environment to compare the suppression effect of the hypha growth between indomethacin and aspirin. (2)Executing the irritation test of indomethacin. on mouse vaginal mucosa (3)Evaluating the suppression effect of the hypha growth in vivo in the estrogen-dependent murine model of candida albicans vaginitis.PartⅠThe Effect of the NSAID Asipirin and Indomethacin on the hypha Growth of Candida AlbicansObjective Comparing the suppression effect of the hypha growth of candida albicans between indomethacin and aspirin under different conditions andchoosing the effective drug and it's concentration that can suppress the hypha growth of candida albicans.Methods Choosing the RPMI-1640 adding 10% calf serum as powerful inducing the hyphal growth condition and chlamydospore test as anoxic environment, indomethacin and aspirin were detected their effect on the hyphal growth and germination of candida albicans at the concentration of 0.5,1,2mmol/L.Results At the level of 0.5,1,2 mmol/L indomethacin and aspirin, the rate of germination were separately 82.58±4.30%,29.25±4.98%,5.08±1.83% and 83.25±2.09%,76.67±1.44%,74.67±1.97%;With comparing control group(0 mmol/L), 0.5,1 mmol/L indomethacin had significant effect on the hypha growth of candida albicans(P<0.05), 2mmol/L indomethacin almost can fully inhibit the hypha growth of candida albicans; 0.5,1,2mmol/L aspirin had no suppression on the hypha growth of candida albicans. In chlamydospore test, 1,2mmol/L indomethacin can valid suppress pseudohypha and chlamydospore growth of candida albicans, and aspirin has no effect on pseudohypha and chlamydospore growth of candida albicans.Conclusion Comparing with aspirin, indomethacin can suppress the hypha growth and germination of candida albicans in powerful inducing the hyphal growth condition and anoxic environment, 2mmol/L indomethacin almost can fully inhibit the hypha growth of candida albicans.PartⅡIrritationTest of Indomethacin on the Mouse Vaginal MucosaObjective To detect the irritation effect and strength of indomethacin on the mouse vaginal mucosa. Methods The experiment sets up blank group and two dosage experiment group. Separately infunding 100μl indomethacin of 2mmol/L,4mmol/L in the mouse vagina of two experiment groups, then repeat administration every 24h for five days, The same treatment for the mice of blank group with normal saline. Sacrificeing the mice after the last administration, cutting the belly open then taking out the complete vagina, cutting it open length wise, observing gross appearance such as hyperemia, oedema, etc. for the reference of pathological checking. Choosing the amphi and the central tissue of the vagina,HE dyeing, then carrying out histopathology examination of vaginal tissue.Results Comparing with the blank group, after infunding 100μl indomethacin of 2mmol/L, 4mmol/L in the mouse vagina of experiment group, we don't observe the irritation reaction of the mouse vaginal mucosa.Conclusion The indomethacin of 2mmol/L, 4mmol/L is safe, no irritation to the mouse vaginal mucosa.PartШMouse Model Test of Indomethacin Inhibiting Hypha Growth of Candida albicansObjective To investigate the therapeutic efficacy of indomethacin on the vaginal candidiasis.Methods The experiment sets up two groups:estrogen-dependent murine model of candida albicans vaginitis as a control group and indomethacin treatment as an experiment group .After Inducing mouse estruation with exogenous estrogen and then infecting candida albicans in the vagina, 100μl indomethacin of 2mmol/L was infunded to the vagina of mice Sequentially for seven days in the treatment group, and so was done 100μl normal saline in the control group.In the second day after the last administration, observing the douche smear with the microscope, counting colony count of Vaginal fungal burden in vagina douche and Sacrificing the mice, executing histopathology examination of vaginal tissue.Results In the indomethacin group there were only a few inflammatory cells (HE dyeing), little hypha, a few spores in vaginal mucosa (PAS dyeing), a low mycetes counting colony in vagina douche and little hypha, a few spores in the douche smear .Conclusion Indomethacin can inhibit hypha growth and Sporogony of candida albicans in Mouse model, and has significantly therapeutical efficacy on the vaginal candidiasis of the mouse.