| Objective:(1)Building a murine model of recurrent vulvovaginal candidias is aim at exploring the metheds of building a recurrent vulvovaginal candidiasis murine model.It lays a foundation of model to research the effective treatment Drugs of recurrent vulvavaginal candidiasis in the clinical by establishing a.recurrent Vulvovaginal candidiasis mouse model.(2)The purpose of detecting the vaginal local immune Factors of recurrent vulvovaginal candidiasis mouse model is to confirm the fuction of vaginal local immune in the pathogenesis of recurrent vulvovaginal candidiasis,provide the basis for the following research.Methods:(1)Firstly,subcutaneously inject with estradiol valerate regulerly for female KunMing mouse to create a state of pseudoestrus,then inoculate Intravaginally with candida albicans in phosphate buffered saline,lastely,a murine model of vulvovaginal candidiasis is established.After doing this,treat them with clotrimazole vaginal tablets(canesten).repeating this procedure three times,then infected with candida albicans successfully which means that a murine model of recurrrent vulvovaginal candidiasis is established successfully.The way to detect this model is observing the vaginal lavage fluid collecting by pbs under the microscope,culturing it by plate it into SDA(sabouraud dextrose ager)after diluting,then obsering the change about its vaginal can be useful.(2)Establishing the recurrent vulvovaginal candidiasis murine model as the methods discribing at the previous part,at the time after infecting each stages successflully,collecting the vaginal lavage except of culture and observing in the microscope then enerlyzing the erxpression about vaginal local immune factors Th1/Th2(IL-8,IL-4)by ELISA(Enzyme linked immunosorbent essay),last analying the data of this experiment by SPSS 17.0 statistical software,the way of it is ANOVA(analysis of variance),P<0.05 was considered as the criterion of significance.Results:(1)The false hyphae can be observed after infected mice at per stages,the results of culture are positive that white vaginal albicans colonies can be founded,in addition,There are some changes in vulva of some mice such as some sectetions and vulva redness,canker.contrary to this,you can not found the false hyphae and the results of culture is negative,then there are no changes you can observe by eyes(2)Analyzing and comparing the expression level of IL-4 and IL-8 at the four stages of establishment of the RVVC mouse.The difference between the expression in the first stage(12.22±2.5)and the second stage(22.2±2.84)of IL-4 was statistically significant,the expression of IL-4 in the first stage is higher than the second stage(P<0.05).the difference between the expression in the first stage(12.22±2.5)and the fourth stage(29.39±5.40)of IL-4 was statistically significant,the expression of IL-4 is apparently higher than the fourth stage(P<0.05).The difference between the expression in the first stage(493.44±68.23)and the third stage(1104.87±306.85)of IL-8 was statistically significant,the expression of IL-8 in the first stage is higher than the third stage(P<0.05).Except of these,the expression of IL-4 in the first stage(12.22±2.5)and in the third stage(43.402±20.19 has no differences.the expression of IL-4 in the second stage(22.29±2.84)and in the third stage(12.22±2.5)has no differences.the expression of IL-4 in the third stage(12.22±2.5)and in the fourth stage(29.39±5.40)has no differences.The expression of IL-8 has no differences in other stages.(3)Analyzing and comparing the expression level of IL-4 and IL-8 in every stage of the estabilishment of RVVC mouse.the difference between the expression of IL-4(12.22±2.5)and IL-8(493.44±68.23)in the first stage was statically significant,and the expression of IL-8 is higher than IL-4 in the first stage(P<0.05).the difference between the expression of IL-4(22.29±2.84)and IL-8(989.323±372.296)in the second stage was statically significant,and the expression of IL-8 is higher than IL-4 in the second stage(P<0.05).the difference between the expression of IL-4(43.402±20.19)and IL-8(1104.87±306.85)in the third stage was statically significant,and the expression of IL-8 is higher than IL-4 in the third stage(P<0.05).the difference between the expression of IL-4(29.39±5.40)and IL-8(773.231±236.086)in the fourth stage was statically significant,and the expression of IL-8 is higher than IL-4 in the fourth stage(P<0.05).ConclusionS:(1)A murine model of recurrent vulvavaginal candidiasis can be established by resonable experiment scheme.(2)Vaginal local immunity is involved in the pathogenesis of vulvovaginal candidiasis.Thland Th2 participate in vaginal local immunity together,after the infection of candida albicans,the ratio of Thl and Th2 changes in the vaginal lavage of the mouse,whicch causes its balance to be damege and the disorder i of vaginal local immunity.That is ralated to the pathogenesis of recurrent vulvovaginal candidiasis possibly. |
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