| Objective:This study is based on the theory of the "V" disease of temperature as the guideline, Using the mixture of Qingyangtoujie, both from clinical studies and animal experiments to observe the relationship between Thl/Th2cell populations offset and imbalance of T lymphocyte subsets in SLE pathogenesis, so as to explore the mechanism of the pathogenesis of SLE, and analyze the mechanism of Qingyangtoujie Therapy.Methods:1. Clinical trial:Adopting prospective, randomized, dingle-blind control trial method, and42patients who were diagnosed as SLE for the first time were enrolled and divided into the treatment group (22cases) and control group (20cases) randomly. In addition, we selected12healthy people as a normal group. Patients in the treatment group were treated with the Qingyangtoujie mixture and western medicine, while those in the control group were treated with placebo and western medicine. Conventional weatern medicine treatment programs are:oral intake of prednisone tablets, add cyclophosphamide if the disease reoccur or lupus nephritis, lupusencephalitis happens. Take venous blood at the beginning of the trial and after6months of treatment to detect the ratio of CD3, CD4and CD8and the number of regulatory T cell of CD4+CD25+, the RBC,WBC,PLT,HGB, complement C3,C4and BUN, Cr; ESR as well.2. Animal experiment:Twenty-four female MRL/lpr mice of8to10weeks were randomly divided into the Chinese Medicine group(group A), the western medicine treatment group(group B), the combined Chinese and western medicine treatment group(group C) and the model control group(group D). Six female BALB/C mice of8to10weeks were selected as a normal control group (group E). Mice in group A were intragastric administrated with mixture of Qingyangtoujie(9.5g/kg),0.4ml per day, group B with prednisone (lOmg/kg),0.4ml per day, group C with prednisone (lOmg/kg), and mixture of Qingyangtoujie(9.5g/kg),0.4ml per day, group D and group E with Sodium Chloride,0.4ml per day. After continuous treatment of six weeks, taking blood to detect the ratio of CD3, CD4and CDS and the number of regulatory T cell of CD4+CD25+. Meanwhile, take the spleen tissue to detect the IFN-rand IL-4gene expression in the spleen with RT-PCR,Result:1. Clinical trial:(1)General information:Thirty-eight cases of the total42cases in the clinical trial were recorded completely, which involves20treatment cases,18control cases and12normal control cases. There are four cases did not complete the final test due to fall off.(2) Clinical parameters:(A)Blood analysis results:There were no significant difference for the RBC level before and after the treatment between the treatment group and the control group (P>0.05), the WBC, the PLT and HGB have no difference before the treatment between these two groups (P>0.05), while there were significant difference between these two groups after the treatment(P<0.05); in the treatment group and control group showed no significant difference after the treatment(P>0.05).(B) Immune parameters:There were no significant difference for the Complement C3, C4level before and after the treatment between the treatment group and the control group (P>0.05). There were significant difference for the treatment group before and after the treatment (P<0.05), while that of the control group has no significant difference (P>0.05). After the treatment, there was significant difference between the treatment group and the control group(P<0.01).(C) Renal function parameters:There were no significant difference for the Urea and creatinine level before the treatment between the treatment group and the control group (P>0.05). There were significant difference for the two groups before and after the treatment (P<0.05). After the treatment, there was significant difference between the treatment group and the control group(P <0.01). (D)ESR results:There were no significant difference for the ESR level before the treatment between the treatment group and the control group (P>0.05). There were significant difference for the two groups before and after the treatment (P<0.05). After the treatment, there was significant difference between the treatment group and the control group(P<0.01).(E) Flow cytometry of CD3+, CD3+CD4+, CD3+CD8+and CD4+CD25+Results: There were significant difference between the treatment group, the control group and the normal control group before the treatment (P<0.01). After the treatment, there were no significant difference between the treatment group and the normal control group(P>0.05), while that of the control group and the normal control group have significant difference (P<0.05). After the treatment, there was no significant difference between the treatment group and the control group (P>0.05). There were significant difference for the two groups before and after the treatment (P<0.05). According to the comparison of the difference before and after the treatment, the effect of the treatment group is relatively better than the control group, and there were significant with the difference (P <0.05).2. Animal experiment:(1)Symptomatic characteristics of the animal:Compared with the normal control group mice, mice in the rest of the treated groups with fur are off and sparse performance, which is more obvious for the model control group performance. The SLE model mice showed tiredness and irritability.(2) RT-PCR test results for the expression of IFN-y mRNA and IL-4mRNA:(A) Thl cytokines IFN-γ gene expression levels:compared with normal group, IFN-γ levels of the model group decreased significantly, there are significant difference (P<0.01), IFN-γ levels of the other treatment groups also decreased with significant difference (P<0.05); model group, the Chinese medicine group, the WM group, treatment group compared with no significant difference (P>0.05).(B)Th2cytokines IL-4gene expression levels:compared with normal group, IL-4level of model group was significantly increased, there are very significant differences (P<0.01), compared with the model group, the Chinese Medicines group, the western medicine group, Chinese and Western medicine combined with group have significant difference (P<0.05), traditional Chinese medicine group, western medicine group, integrative medicine group, normal group, pairwise comparisons showed no significant difference (P>0.05).(C)Of Th1/Th2cells factor ratio:Compared with normal group, the TCM group, model group have very significant differences (P<0.01), Chinese and Western medicine combined with group and normal group has no obvious significant difference (P>0.05), WM group compared with normal group has significant difference (P<0.05); there are significant differences among each treatment groups and the model group(P<0.05).(3) Flow cytometry analysis of the T lymphocyte subset level:(A) CD3+level: Compared with normal group, CD3+levels of model group mice decline, there are significant differences (P<0.01), compared with model group, CD3+levels in all treatment groups rise, there are significant differences (P <0.05). Traditional Chinese medicine group and western medicine group, combined with western medicine group, and normal control group pairwise comparisons was not significant (P>0.05).(B) CD3+CD4+levels:compared with normal group, CD3+CD4+levels of the model group obviously decreased, there was significant difference (P<0.01), traditional Chinese medicine group and western medicine group, Chinese and Western medicine combined group no significant difference (P>0.05).(C) CD3+CD8+level:compared with normal group, model group CD3+CD8+levels rise, there was a significant difference (P<0.01), traditional Chinese medicine group and western medicine group, Chinese and Western medicine combined group no significant difference (P>0.05).(D) CD4+CD25+levels:Compared with normal control group, mouse peripheral blood CD4+CD25+in the model group and each treatment groups have significant difference (P<0.01); there were significant differences between the model group and Integrative Medicine group (P<0.01), the WM group and Integrative Medicine group, that of the Western medicine and the traditional Chinese medicine group pair wise comparisons were no significant differences (P>0.05).Conclusion:1. According to the result of clinical experiment: The experimental data can indicated that the SLE patients showed balance disorder of the T lymphocytes, which expressed as the increase of CD3+, decrease of CD3+CD4+, and increase of CD3+CD8+and CD4+CD25+, and indicated Th1/Th2balance disorders of Th1/Th2cells drift to Th2. The mixture-based treatment group was superior to the simple use of western medicine in the control group, and Integrative Medicine treatment group can improve the Thl/Th2cell population imbalance, maintaining normal T cell function, inhibiting the abnormal proliferation of B cells.2. Experiment on animal showed that:MRL/lpr mice have balance disorder of the Th1/Th2, mainly showed as Th2increased, Th1/Th2cells drift to Th2as well as the decrease of T lymphocyte subsets and regulatory T cells. The effect of the integrated treatment group is obviously better than the other treatment group.3. There was difference in the results of experimental data and clinical manifestations:RBC before and after treatment in the blood of clinical patients has no significant difference, while during clinical, some patients' abnormal condition of the RBC turned to normal after a period time of treatment, which indicating that there is still change in the trend, but just due to the sample size and time constraints led to no significant statistical difference.
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