Study On The Immunomodulatory Mechanism Of Wenjinghuayu-zhitong Treatment On T Cell Subsets Differatiation In Primary Dysmenorrhea Of The Type Of Hanning Blood Stasis And Medication Principle Of Primary Dysmenorrhea

Objective:To investigate the effect and mechanism of Wenjinghuayuzhitong treatment on the differatiation of T cell subsetsand regulation of cytokines balance in rats with primary dysmenorrhea of syndrome of blood stasis due to cold accumulation.Meanwhile,systematically analyze the medication principle of primary dysmenorrhea based on the literature of modern TCM clinical study on primary dysmenorrheaMethods:In the clinical experimental investigation,the rat model of primary dysmenorrhea of syndrome of blood stasis due to cold accumulation was induced by oxytocin and ice at zero degree.All the 50 rats were randomly divided into normal group,model group,low does group,high dose group and ibuprofen group.Each group was given drugs for six consecutive days except the normal group and injected oxytocin 30 minutes after last administration,for the purpose to observe the frequency of writhing response in 30 minutes.Thequantity of T cell subsets in rats’ spleens mononuclear were measured by flow cytometry,the level of IFN-γ,IL-4,IL-17 and TGF-β were measured by ELISA,and the mRNA expressions of TNF-α,IL-4,IL-17A and TGF-β were assessed by RT-PCR.In the modern TCM literature stuty,a database of recipes and herbs was founded with computer technology.Then analyzed the medication principle from the point of frequency analysis,association rules and varable clustering analysis.Results:Compared with normal group,the percentage of Th2 and Th17 cells in model groupwere increased while the percentage of Thl and Treg cells were decreased significantly(P<0.01),and the contents of IFN-γ and TGF-β and the mRNA expression of TNF-α and TGF-β were decreased while the contents and the mRNA expression of IL-4 and IL-17 were increased significantly(P<0.01),which notes primary dysmenorrhea may exist the Thl to Th2 shift and Treg to Th17 shift.Compared with model group,the latency period of writhing response was extended and the frequency of the writhing response was less in low dose and high dose groups(P<0.05).Additionally,the percentage of Th1 and Treg cells in drug groups were increased while the percentage of Th2 and Thl 7 cells weredecreased significantly(P<0.05,P<0.01),the contents of IFN-γ and TGF-β and the mRNA expression of TNF-α and TGF-β in drug groupswere increased significantly(P<0.01),while the contents of IL-4 and IL-17 and the mRNA expression of IL-4 and IL-17A were decreased markedly(P<0.01).There are 221 primary dysmenorrhea clinical study literatures with 126 types of herbs are used.The ratios of the tonic and blood circulation drugs were highest among all.According to the frequency analytic method,the blood circulation drugs were highest with the rate of 31.46%,followed by the tonic drugs with the rate of 28.81%.The ratio of the herb with the nature of warm was highest while the ratio of the herb with flavors of spicy,sweet and bitter was the highest.Concerning the Zang Fu meridian belonging of the herb,most herbs belong to the Liver,Spleen and Kidney lines.According to the association rules,the core drug is Danggui.With the varable clustering analysis,three basic formula was formed.Conclusions:The imbalance of the T cell subsets differatiation was involved in rats with primary dysmenorrhea of syndrome of blood stasis due to cold accumulation.Xiang Yan Zhi Tong Prescription relieves symptoms by regulating Thl/Th2 and Th17/Treg cytokines balance.The treatment principle of primary dysmenorrhea in TCM is based on tonifying kidney and nourishing qi and blood,while activating blood for easing pain.