Study On The Effect Of Lisea Cubeba Oil Against Mice Model Of Candidal Vaginitis

Objective: To observe the effect of Lisea Cubeba Oil against mice model of candidal vaginitis, whether it possessed the effect of treating and anti-pruritus or not, this was studied by constructing mice model of Candidal Vaginitis. We also sieved the concentration which could be external application to treat Candidal Vaginitis. The study can provide experiment data for clinical application of Lisea Cubeba Oil.Metheods:1 Mice model of Candidal Vaginitis:Six weeks old, ten healthy female Kunming mice were used for study. Six days before inoculation, mice were injected subcutaneously with Estradiol Benzoate 0.1ml every other day in order to make the mice be in the state of pseudoestrus, until the study was finished. On the inoculation day, the mice were inoculated with 5×10~4 stationary-phase blastoconidia of Candidia albicans intravaginally and five vaginal tissues were obtained on day 2,4 after inoculation. The mice were taken the samples by swabs before being sacrificed. The fungus was tested under microscope.2 The stimulation test:Six weeks old, fifty healthy female Kumming mice were used for study. Mice were randomized into five groups: groupⅠ,groupⅡ, groupⅢ,groupⅣand groupⅤ,10 in each group. GroupⅡ,groupⅢ,groupⅣ,groupⅤwere dealt with 4% ,8%,12% and 16% of Litsea Cubeba Oil respectively. GroupⅠwas dealt with sodium chloride. The treatment of each group was the same:all drugs were administered once daily for a total of 7 days, 30μl every time. The food and drink, action, mental status, dejecta and so on were monitored every day among 7 days. 24 hours after the last administer, animals were sacrificed and the vaginal tissues were obtained.3 Therapeutic action test:45 mice model of Candida albicans vaginitis were randomized into five groups: group A, group B, group C, group D and group E, 9 in each group. Group B, group C, group D were dealt with 4%, 8%, 12% Litsea Cubeba Oil. Group A was dealt with sodium chloride and group E was dealt with Fluconazol. The treatment of each group was the same:all drugs were administered once daily for a total of 7 days, 30μl every time. 50μl PBS fluid were used to irrigation vagina 8 to 10 times on day 0, 5, 7 and 21 after treatment. The vaginal lavage fluid were mixed and serially diluted 1:10. 20μl fluid were plated on SDA for 24 h at 35°C.4 Anti-pruritus test: 30 healthy guinea pigs were used for this study. The hair of the Right-after foot back was cutted. Every other day, the place where the hair was cutted was abrasioned by coarse sand paper. The area of the place was about 1cm2. Those guinea pigs were randomized into five groups:group jia, group yi, group bing, group ding and group wu, 6 in each group. Group yi, group bing, group ding were dealt with 4%, 8%, 12% Litsea Cubeba Oil and group jia was dealt with sodium chloride. Dosage of drugs was 1ml every animal. Every animal of group E was given 0.05g Hydrocortisone Butylate. 30 minutes after partly treated, the abrasioned place were besmeared by 0.05% histamine phosphate 50μl. Among 3 minutes if the guinea pig did not lick its abrasioned place, repeated it until the action was occurred. When the guinea pig licked its abrasioned place, we recorded histamine phosphate's frequency and dosage.All the data were analyzed by SPSS11.5. The results were statistically processed by Oneway Anova. P<0.05 was significant.Results:1 Mice model of Candidal Vaginitis:There were many vaginal epithelial cells and sporus under the microscope on the second day after inoculation. Many sporus were found in the superficial mucosa of vaginal tissue. On the fourth day, a flock of vaginal epithelial cells, sporus and hypha were founded. There were large amount of hypha and sporus in the superficial mucosa of vaginal tissue.2 The stimulation test:Within the 7 days, the food and drink, action, mental status, dejecta and so on were normal. No animal was dead. Compared with groupⅠ, there was no obviously difference. The result of histopathology: the layer of vaginal tissue cuticular layer of 4 groups(groupⅠ, groupⅡ, groupⅢ, groupⅣ)were clear and monolayer tactic stylolitic basal cell can be seen. Blood vessel was not obviously dilated and congestive. Connective tissue was light oedema. A little phlegmonosis cells were penetrated. The mice's vaginal mucosa of groupⅤwere intracellular edema of epidermis. The cells of the prickle cell layer were obviously vacuolus degeneration and basal cells were liquefaction degeneration. Blood vessel was dilated and congestive obviously. Connective tissue was oedema. There were more phlegmonosis cells invade. It indicated that 16% Litsea Cubeba Oil stimulate the mice's vaginal mucosa obviously that could not be used to treat the candidal vaginitis.3 The results of the therapeutic action test: The vaginal douche colony count logarithm counts of group A, group B, group C, group D and group E were 7.42±0.28, 6.94±0.40, 6.52±0.42, 6.11±0.27, 5.31±0.48, respectively, after treated 7 days. All concentration of Litsea Cubeba Oil and fluconazol could degrade the average of colony count logarithm counts. There was significant deviation compared with group A (P<0.05). Furthermore, with the increase of the Litsea Cubeba Oil's therapeutic concentration, the colony count logarithm counts were descending. There was statistical significance (P<0.05).4 The results of anti-pruritus test:Different concentrations of Litsea Cubeba Oil and Hydrocortisone Butylate all could raise the functions of histamine phosphate's itch-threshold to guinea pig. Compared with group jia,there was statistical significance(P<0.05). The reactive of different concentrations of Litsea Cubeba Oil to histamine phosphate's itch-threshold increased with the increase of the drugs. There was statistical significance (P<0.05).Conclusion:1 The stimulation of 4%,8%,12% Litsea Cubeba Oil is poor. They all can be used to short-term treatment of Candidal Vaginitis.2 Litsea Cubeba Oil can be external application to treat candidal vaginitis. With the raise of the drug's concentration, therapeutic effect is increased.3 Litsea Cubeba Oil has the anti-pruriti function caused by histamine phosphate.