| Objective:To establish an composite animal model of genital chlamydia trachomatis(Ct as brief)infection and anti-sperm antibody (Asab as brief) positive SD rats by the means of simulating the natural onset of human being. To investigate the change of general immune function on genital chlamydia trachomatis infection and Asab positive SD rats as well as the influence of "QingShiHuaYu Mixture" (QSHY as brief). To further explore the pathogeny of anti-sperm immune infertility and the possible mechanism of the Chinese medicine therapy. Then simultaneously, to study the distribution of chlamydia trachomatis infection and Asab positive in infertile women and the relationship with the distribution of TCM syndrome. To offer an clinic evidence by exploring the relationship between chlamydia trachomatis infection, AsAb and infertility and the influencing factors. Methods and Results:1. Clinical research:178 women with infertility were evaluated and made the syndrome differentiation. The AsAb in serum samples and Ct of cervical secretion of 178 women with infertility and that of 25 healthy women were detected. To know the coherence and influence factors of chlamydia trachomatis infection and the AsAb, and then analyse the relationship between the distribution of TCM syndrome and the AsAb. Results:①The AsAb positive rate of infertility group was 36.52%, while 31.33% of the primary infertility group and 41.05% of the secondary infertility group, which were all obviously higher than the control group (P<0.05). The infection rate of Ct was 21.91%, which was 18.07% of the primary infertility group and 25.26% of the secondary infertility group, both of them were also higher than the control group (P <0.05). The infection rate of Ct was higher among those women with the AsAb positive and the Ct has significant positive correlation with the AsAb. As the abortion times increased, both the infection rate of Ct and the AsAb positive rate were going up. There was significant difference between the group of more than three times of abortion with the group of less than three times (P<0.05).Compared with the group of less than three years, the infection rate of Ct raised (P<0.05) while the AsAb positive rate without change in the group at least three years'infertility history.②Among the 178 infertility women, the hot and humid stasis resistance syndrome distributed most and taken up 34.3%, followed the nephrasthenia and blood stasis syndrome which taken up 28.09%; and the least was the liver-kidney Yin deficiency syndrome. Most of the patients age is from 25 to 35, and the hot and humid stasis resistance syndrome and the nephrasthenia and blood stasis syndrome are show on that time. The hot and humid stasis resistance syndrome group had the highest rate of AsAb positive and Ct infection. The AsAb positive rate was highest in patient with the hot and humid stasis resistance syndrome and the nephrasthenia and blood stasis syndrome secondly. The AsAb impacted on the distribution of the hot and humid stasis resistance syndrome and the nephrasthenia and blood stasis syndrome, while the Ct impacted on the distribution of the hot and humid stasis resistance syndrome.2. Experiment research:Ct E serotype of clinical strains were cultured in McCoy cells and used to build the rat animal model of genital Ct infection. To establish an composite animal model of genital Ct infection and Asab positive SD rats by the means of simulating the clinic pathologic course. The SD rat were randomly divided into five groups, these were the pure Ct infection group (A), the pure Asab immunity group (B), the Ct plus Asab treatment group (C), the Ct plus Asab model group(D) and the blank control group(E). According to different models, the rats were injected Ct E serotype or/and human being's sperm antigen through vagina, and the treatment group was treated with QSHY through stomach for three weeks. The level of AsAb,interleukin 6 and interferon Y in serum samples were both detected before and after the experiment. Ct of the cervical secretion was detected by cultivation when the models were set up. After the experiment the rats were killed to check the content of CD4+,CD8+,NK of splenic lymphocytes in rats. Results:①The weight difference of the models was significantly differed between group A and B,C and D,D and E (P<0.01), which pointed out that the weight of the Ct plus Asab model group was much lighter than the control group.the Ct plus Asab model rats became fretful and had acute inflammation symptoms like increasing vaginal secretion and vulva swollen at the beginning, and later the secretion became thick, the bowels became loose, exercise and appetite reduced et al. Compared with before-molding, the titer of Asab all raised in the pure Ct infection group, the pure Asab immunity group and the Ct plus Asab model group which was the highest one. There was significant difference before and after molding both in group B and group D (P<0.01). There was also significant difference between the groups, thus as group B to E, group D to E, and group B to D (P<0.01)②The level of serum IL-6 of the pure Ct infection group and the Ct plus Asab model group all increased comparing with the control group, but without significant difference (P<0.05).The level of serum IFN-γof the pure Ct infection group, the pure Asab immunity group and the Ct plus Asab model group all increased, and the Ct plus Asab model group was higher than the other two groups (P<0.01). The level of serum IFN-γbetween the pure Ct infection group and the pure Asab immunity group was not significantly different (P>0.05) The content of CD4+,CD4+/CD8+,NK of splenic lymphocytes in rats all raised in the pure Ct infection group, the pure Asab immunity group and the Ct plus Asab model group, and the Ct plus Asab model group was higher than the other two groups (P<0.01). there was significantly difference between all the three groups with the control group (P<0.05).The content of CD8+ of splenic lymphocytes was decreased both in the pure Asab immunity group and the Ct plus Asab model group, and the Ct plus Asab model group was lower than the control group significantly (P<0.01).③The serum AsAb titer has significant positive correlation with the level of serum IFN-γ, the content of CD4+,CD4+,CD8+,NK of splenic lymphocytes (P<0.01), while negative correlation with the content of CD8+. The level of serum IFN-γalso has significant positive correlation with the content of CD4,CD4+/CD8,NK of splenic lymphocytes (P<0.01).④The symptoms of the Ct plus Asab model with herbal medicine group were improved. And the weight increased obviously compared with the Ct plus Asab model group (P<0.01). The serum AsAb titer of the treatment group has significantly descended than the model group (P<0.01), and there was no significant difference with the control group (P>0.05). As the content of splenic lymphocytes, CD4+,CD4+,CD8+,NK of the rats with herbal medicine all decreased while the CD8+ increased than the Ct plus Asab models. The difference between them were significantly (P<0.01).The level of serum IL-6 of the treated group was lower than the control group, but the difference was not significant (P>0.05).The level of IFN-γof the treated group was decreased obviously than the Ct plus Asab model group (P<0.01),while there was no significant difference with the control group (P>0.05)Conclusion:1. Infection of Ct and Asab all can induce female infertilily. The production of Asab may be related to the infection of Ct. The infection of Ct and its induction of AsAb may be one of the cause of female immune infertilily. The hot and humid stasis resistance syndrome and the nephrasthenia and blood stasis syndrome is the main type of infertility. Both infection of Ct and AsAb have significant correlation with The hot and humid stasis resistance syndrome.2. Successed for Ct E serotype of clinical strains on proliferation cultivation, and established SD rats chlamydia trachomatis infection model through vaginal inoculation method with Ct E serotype of clinical strains. The antisperm immune model that based on chlamydia trachomatis infection conforms to the clinical practice.3. Chlamydial trachomatis infection may cause active sperm immune response and that will induce the generation of Asab. Both chlamydia trachomatis infection and antisperm antibody can cause the immune function disorder, and they have synergistic effect. The Ct plus Asab model rats existed imbalance of Thl/Th2 and Th/Ts.4. QingShiHuaYu Mixture could adjust the content of lymphocytes and cytokines, improve the immunologic function of the antibody genital Ct infection and Asab positive SD rats, thus reduce and eliminate serum Asab, so as to achieve the purpose of improving reproductive function. |
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