基于沙眼衣原体感染的抗精子免疫模型及中药干预和免疫功能变化
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摘要
目的：本研究通过模拟人抗精子免疫性不孕的发病途径,建立女性生殖道衣原体感染抗精子免疫复合模型,研究沙眼衣原体感染和抗精子免疫状态下全身免疫功能的变化,以及中药清湿化瘀合剂的干预作用,进一步探讨抗精子免疫性不孕的发生机理,以及中医药治疗本病的作用机理。同时通过临床调查,了解沙眼衣原体感染和抗精子抗体阳性不孕患者在不孕人群中的分布情况,及与中医证型分布的关系,探讨沙眼衣原体、抗精子抗体和不孕的关系和相关影响因素,为本研究提供临床佐证。
方法和结果：
1.临床研究：制定临床观察表,对178例不孕患者进行中医辨证分型,同时检测不孕患者和正常对照组血清抗精子抗体定量和宫颈沙眼衣原体感染情况,了解衣原体感染和抗精子抗体的相关性和影响因素。分析不孕症中医证型与AsAb、Ct感染的关系。结果：①不孕组中AsAb阳性率36.52%,其中原发性不孕组AsAb阳性率31.33%,继发性不孕组AsAb阳性率41.05%,均明显高于对照组(P<0.05)。Ct感染发病率为21.91%,其中原发性不孕者15例,占18.07%,继发性不孕组24例,占25.26%,均明显高于正常对照组(P<0.05)。AsAb阳性患者中衣原体感染率高于AsAb阴性组(P<0.05), Ct与AsAb有显著正相关性。随着流产次数增加,AsAb阳性率和Ct感染率均升高,流产次数≥3次组与流产次数<3次组相比,有显著性差异(P<0.05)。不孕病程3年以上组与病程3年内组相比,Ct的感染率上升(P<0.05),而AsAb阳性率无明显变化。②178例不孕患者中,湿热瘀阻型分布最多,其构成比为34.3%；肾虚血瘀型则占28.09%；肝肾阴虚型最少,占17.42%。不孕患者年龄最多见于25～35岁之间,且此年龄段主要表现为湿热瘀阻证和肾虚血瘀证。湿热瘀阻型组抗精子抗体阳性率和Ct感染率均最高。抗精子抗体与湿热瘀阻型和肾虚血瘀型关系最密切,而Ct感染则形成湿热瘀阻证的可能性最大。
2.实验研究：增殖培养E型沙眼衣原体临床株,建立沙眼衣原体生殖道感染大鼠模型。模拟临床病理过程,采用生殖道局部Ct感染和免疫的方法,建立衣原体感染抗精子抗体免疫大鼠模型：随机将SD大鼠分为5组,A组(单纯炎症模型+生理盐水组)、B组(单纯免疫模型+生理盐水组)、C组(炎症免疫模型+清湿化瘀合剂组)、D组(炎症免疫模型+生理盐水组)、E组(空白对照+生理盐水组)。用阴道接种的方法,分别按不同模型接种CtE型原体和/或人精子抗原,并对治疗组予清湿化瘀合剂灌胃治疗3周。实验前后检测血清AsAb、IL-6、IFN-y水平,造模后用培养法检测Ct感染情况,实验后取大鼠脾脏淋巴细胞检测CD4+、CD8+、NK细胞含量。结果：①造模前后体重差A组与D组、C组与D组、D组与E组相比均有显著差异(P<0.01)。说明Ct感染、抗精子免疫和复合模型体重增长均较空白组慢,D组复合模型组最明显。复合模型组大鼠开始时出现烦躁、阴道分泌物增多,外阴红肿等急性炎症表现,后期出现分泌物变稠、大便稀烂、活动纳食减少及松毛等现象。与造模前相比,A组(单纯Ct感染)、B组(单纯免疫)、D组(Ct感染+生殖道主动免疫)抗精子抗体滴度均升高,其中以复合模型组升高最明显。其中B组、D组各自造模前后比较差异有显著意义(P<0.01)。造模后B组、D组与E组(空白对照)相比,差异有显著意义(P<0.01)B组与D组组间相比,差异亦有显著意义(P<0.01)。②衣原体感染组,复合模型组大鼠外周血IL-6与空白对照组相比有所升高,但差异无显著意义(P<0.05)。造模后单纯炎症组、单纯免疫组、及复合模型组血清IFN-γ均升高,复合模型组升高明显,与其他两组有显著性差异(P<0.01),而单纯炎症及单纯免疫组组间无明显差异(P>0.05)。单纯免疫组及复合模型组均出现脾脏NK细胞的含量升高,且复合模型组升高最明显,与正常对照组相比有显著性差异(P<0.05)。单纯Ct感染、单纯免疫模型及Ct感染抗精子免疫复合模型组的CD4+升高及CD4+/CD8+均明显升高,与正常对照组对比,差异有显著性(P<0.05),其中以复合模型组升高最明显。单纯免疫组和复合模型组CD8+含量均下降,其中复合模型组与正常对照组相比有显著性差异(P<0.01)。③血清AsAb定量与外周血清IFN-γ、脾脏CD4+含量、CD4+/CD8+比率、NK细胞含量都有显著的正相关性(P<0.01),而与CD8+细胞含量则呈负相关性。血清IFN-γ水平与脾脏CD4’、CD4+/CD8+、NK细胞均有显著正相关性(P<0.01)。④中药治疗后,治疗组大鼠阴道分泌物减少,外阴红肿情况消失,嗜睡、大便、松毛及活动减少等情况均明显好转。体重增加较复合模型组明显(P<0.01)。与模型组对比,治疗组外周血清AsAb定量明显下降(P<0.01)。与正常组相比,P>0.05。在淋巴细胞方面,与复合模型组相比,治疗组大鼠脾脏NK细胞、CD4+、CD4+/CD8+比率均下降,而CD8+表达升高,差异均有显著意义(P<0.01)。治疗组血清IL-6较复合模型组有所下降,组间比较无显著性差异(P>0.05)。C组大鼠外周血IFN-γ水平明显下降,与正常组差异无显著性(P>0.05),与D组模型组有显著差异(P<0.01)。
结论：
1.Ct感染及AsAb均可造成女性不孕,Ct感染本身及诱发AsAb产生双重因素可能是女性免疫性不孕的原因之一。湿热瘀阻证和肾肾虚血瘀证是不孕的主要证型,抗精子抗体和衣原体感染均与湿热瘀阻证有显著相关性。
2.成功对沙眼衣原体E血清型临床株进行增殖培养,并通过阴道接种的方法建立SD大鼠沙眼衣原体感染模型。基于沙眼衣原体感染的抗精子免疫模型更符合临床实际。
3.衣原体感染后的精子主动免疫应答可能诱发抗精子抗体的产生,衣原体感染和抗精子抗体都能引起机体免疫功能的紊乱,且两者有协同作用。衣原体感染抗精子免疫大鼠模型存在Th1/Th2、Th/Ts的平衡失调。
4.清湿化瘀合剂能够调节淋巴细胞和细胞因子的含量,改善沙眼衣原体感染抗精子免疫大鼠模型的免疫功能,降低抗精子抗体定量,从而改善生殖功能。
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.
方法和结果：
1.临床研究：制定临床观察表,对178例不孕患者进行中医辨证分型,同时检测不孕患者和正常对照组血清抗精子抗体定量和宫颈沙眼衣原体感染情况,了解衣原体感染和抗精子抗体的相关性和影响因素。分析不孕症中医证型与AsAb、Ct感染的关系。结果：①不孕组中AsAb阳性率36.52%,其中原发性不孕组AsAb阳性率31.33%,继发性不孕组AsAb阳性率41.05%,均明显高于对照组(P<0.05)。Ct感染发病率为21.91%,其中原发性不孕者15例,占18.07%,继发性不孕组24例,占25.26%,均明显高于正常对照组(P<0.05)。AsAb阳性患者中衣原体感染率高于AsAb阴性组(P<0.05), Ct与AsAb有显著正相关性。随着流产次数增加,AsAb阳性率和Ct感染率均升高,流产次数≥3次组与流产次数<3次组相比,有显著性差异(P<0.05)。不孕病程3年以上组与病程3年内组相比,Ct的感染率上升(P<0.05),而AsAb阳性率无明显变化。②178例不孕患者中,湿热瘀阻型分布最多,其构成比为34.3%；肾虚血瘀型则占28.09%；肝肾阴虚型最少,占17.42%。不孕患者年龄最多见于25～35岁之间,且此年龄段主要表现为湿热瘀阻证和肾虚血瘀证。湿热瘀阻型组抗精子抗体阳性率和Ct感染率均最高。抗精子抗体与湿热瘀阻型和肾虚血瘀型关系最密切,而Ct感染则形成湿热瘀阻证的可能性最大。
2.实验研究：增殖培养E型沙眼衣原体临床株,建立沙眼衣原体生殖道感染大鼠模型。模拟临床病理过程,采用生殖道局部Ct感染和免疫的方法,建立衣原体感染抗精子抗体免疫大鼠模型：随机将SD大鼠分为5组,A组(单纯炎症模型+生理盐水组)、B组(单纯免疫模型+生理盐水组)、C组(炎症免疫模型+清湿化瘀合剂组)、D组(炎症免疫模型+生理盐水组)、E组(空白对照+生理盐水组)。用阴道接种的方法,分别按不同模型接种CtE型原体和/或人精子抗原,并对治疗组予清湿化瘀合剂灌胃治疗3周。实验前后检测血清AsAb、IL-6、IFN-y水平,造模后用培养法检测Ct感染情况,实验后取大鼠脾脏淋巴细胞检测CD4+、CD8+、NK细胞含量。结果：①造模前后体重差A组与D组、C组与D组、D组与E组相比均有显著差异(P<0.01)。说明Ct感染、抗精子免疫和复合模型体重增长均较空白组慢,D组复合模型组最明显。复合模型组大鼠开始时出现烦躁、阴道分泌物增多,外阴红肿等急性炎症表现,后期出现分泌物变稠、大便稀烂、活动纳食减少及松毛等现象。与造模前相比,A组(单纯Ct感染)、B组(单纯免疫)、D组(Ct感染+生殖道主动免疫)抗精子抗体滴度均升高,其中以复合模型组升高最明显。其中B组、D组各自造模前后比较差异有显著意义(P<0.01)。造模后B组、D组与E组(空白对照)相比,差异有显著意义(P<0.01)B组与D组组间相比,差异亦有显著意义(P<0.01)。②衣原体感染组,复合模型组大鼠外周血IL-6与空白对照组相比有所升高,但差异无显著意义(P<0.05)。造模后单纯炎症组、单纯免疫组、及复合模型组血清IFN-γ均升高,复合模型组升高明显,与其他两组有显著性差异(P<0.01),而单纯炎症及单纯免疫组组间无明显差异(P>0.05)。单纯免疫组及复合模型组均出现脾脏NK细胞的含量升高,且复合模型组升高最明显,与正常对照组相比有显著性差异(P<0.05)。单纯Ct感染、单纯免疫模型及Ct感染抗精子免疫复合模型组的CD4+升高及CD4+/CD8+均明显升高,与正常对照组对比,差异有显著性(P<0.05),其中以复合模型组升高最明显。单纯免疫组和复合模型组CD8+含量均下降,其中复合模型组与正常对照组相比有显著性差异(P<0.01)。③血清AsAb定量与外周血清IFN-γ、脾脏CD4+含量、CD4+/CD8+比率、NK细胞含量都有显著的正相关性(P<0.01),而与CD8+细胞含量则呈负相关性。血清IFN-γ水平与脾脏CD4’、CD4+/CD8+、NK细胞均有显著正相关性(P<0.01)。④中药治疗后,治疗组大鼠阴道分泌物减少,外阴红肿情况消失,嗜睡、大便、松毛及活动减少等情况均明显好转。体重增加较复合模型组明显(P<0.01)。与模型组对比,治疗组外周血清AsAb定量明显下降(P<0.01)。与正常组相比,P>0.05。在淋巴细胞方面,与复合模型组相比,治疗组大鼠脾脏NK细胞、CD4+、CD4+/CD8+比率均下降,而CD8+表达升高,差异均有显著意义(P<0.01)。治疗组血清IL-6较复合模型组有所下降,组间比较无显著性差异(P>0.05)。C组大鼠外周血IFN-γ水平明显下降,与正常组差异无显著性(P>0.05),与D组模型组有显著差异(P<0.01)。
结论：
1.Ct感染及AsAb均可造成女性不孕,Ct感染本身及诱发AsAb产生双重因素可能是女性免疫性不孕的原因之一。湿热瘀阻证和肾肾虚血瘀证是不孕的主要证型,抗精子抗体和衣原体感染均与湿热瘀阻证有显著相关性。
2.成功对沙眼衣原体E血清型临床株进行增殖培养,并通过阴道接种的方法建立SD大鼠沙眼衣原体感染模型。基于沙眼衣原体感染的抗精子免疫模型更符合临床实际。
3.衣原体感染后的精子主动免疫应答可能诱发抗精子抗体的产生,衣原体感染和抗精子抗体都能引起机体免疫功能的紊乱,且两者有协同作用。衣原体感染抗精子免疫大鼠模型存在Th1/Th2、Th/Ts的平衡失调。
4.清湿化瘀合剂能够调节淋巴细胞和细胞因子的含量,改善沙眼衣原体感染抗精子免疫大鼠模型的免疫功能,降低抗精子抗体定量,从而改善生殖功能。
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.
引文
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