The Experimental Research Of Zengye Bujin Decoction For Intervention Effeets Of Sjogren’s Syndrome Model Mice

Objetctives:To establish experimental Sjogren’s syndrome (SS)model by induced with immune inducement method and observe dynamicly the effect of Zengye Bujin Decoction on symptoms and immune function of reguLating of mice model, from the point of molecuLar cell biology to explore the influence of TNF-a, IL-6, aquaporin-5with Zengye Bujin Decoction, to provide basic evidence for its mechanism, make its therapeutic targets clearly, at the same time provide the experiment evidence for better use it to the clinic.Method:1. Method of theoretical researeh:By retrieving literature at home and abroad,from Ancient to present,summarized the traditional Chinese medicine clinical syndrome differentiation, therapeutic methods and drugs and experimental study of SS.2. Method of experimental research:BALB/c rats were divided into normal group, model group, Zengye Bujin Decoction low dose group, Zengye Bujin Decoction middle dose group, Zengye Bujin Decoction high dose group, Anethol Trithione group.In addition to normal group, other five groups were induced with immune inducement method(injection of Pertussis vaccine, complete Freund’s adjuvant and mice glanduLa anguLaris antigen).Then the normal group, model group was given normal saline gavage, Zengye Bujin Decoction low dose (6.8g. kg-1), middle dose(12.6g. kg-1), haigh dose(27.3g. kg-1) was given treatment group, and Anethol Trithione(12.5mg.kg-1)was given positive group. After first immune, to reinforce immune with emuLsifier antigens in third, seventh days, the dose is0.2mL, sc on back with more points. At intervals of14days, to reinforce immune with emuLsifier antigens first, the dose and position as same as the previous twice, total of two times to reinforce immune to the end of experiment. Normal group has not deal anything. In50days of experiment, to observe the effect of Zengye Bujin Decoction on general indicators(Weight, water intake, food intake, saliva flow rate).When experiment was finished, to take Submaxillary gland. Thymus gland, spleen gland, calcul.ate Submaxillary gland index. Thymus gland index, spleen gland index, to observe pathological changes of Submaxillary. To take blood from eyes, serum from centrifugal, cytokine TNF-α and IL-6in the mice with SS was shown by ELISA analysis, to detect the AQP5expression changes of the submaxillary gland cells in mice through Western blots.ResuLts:1. ResuLts of theoretical research:Based on the analysis of literature, we think of various doctors of Chinese medicine make diagnosis and treatment from different point of view, but revolves around the pathological character of difficiency in origin and excess in superficiality to treatment based on differential diagnosis. Dificiency in origin referred to Qi-blood and Yin-Yang lost of every Zang-fu organ, the most important of it is that fluid exhaustion, Zang-fu organ malnutrition, disturbance of fluid in transportation, while excess in superficiality referred to cementation of toxin and blood stasis. So according to the theory of Qi-blood-fluid, draft the Zengye Bujin Decoction to invigorating Qi and nourishing Yin, distributing fluid and free collaterals, promoting blood circulation to remove blood stasis to treat SS.2. Experiment research resuLts2.1General state performance:Compared with the normal group, model group salivary flow rate decreased, water intake increased, food intake and weight reduced. In addition to, normal group in good shape, activities freely, fur smooth, not mouLt; model mices Spirits are drooping, sleepiness, lazy, mobility decreased, the fur glossiness droupped, loosen indistinctly and tail uLceration and crust of some mices, while each dose group of Zengye Bujin Decoetion can improve the symptoms of deficiency Qi-Yin, mouLt slightly and fur loosen not obvious.2.2Observe form and structure changes of submandibuLar gland by microscope: submandibuLar organization have massively lymphocytic infiltration, acinar atrophy or disappear, duct expansion etc, submandibuLar atrophy, submandibuLar index decreased, while Zengye Bujin Decoction can reduced lymphocytic infiltration, inhibit it atrophy.2.3Immunological function:Compared with the normal group,Thymus gland index of model group increased, spleen gland index increased(P<0.01, P<0.05), explain strong immune response happened in body, so infer that Zengye Bujin Decoction couLd suppress the immune response.2.4Changes of cytokincTNF-α. II-6:Compared with the normal group, TNF-a and IL-6of model group raised(P<0.01):compared with the modle group, different dosage of Zengye Bujin Decoetion couLd reduced TNF-α, IL-6(P<0.01).2.5ResuLt of Western blot:Compared with the normal group, the expression of AQP5declined27.90%(P<0.01); compared with the model group, the high, middle, low dose group of Zengye Bujin Decoction couLd raised the expression of AQP532.84%(P<0.01),12.54%(P<0.05),4.51%, Anethol Trithione group raised20.40%(P<0.01).Conclusion:Zengye Bujin Decoction couLd inhibit secretion of cytokineTNF-a, IL-6, indirectly to suppress the helper T cells (Th) proliferation and differentiation to reduce the immune inflammation reaction, relive the Iymphocytic infiltration on submaxillary gland, structure of acinus, ductal and immune organ damage, protection saliva secretion of submandibuLar, promote AQP5expression, show that cytokine and AQP5are play an important role in the course of SS furtherly, at the same time, prove the global reguLatory role of invigorating Qi and nourishing Yin, distribute fluid and free collaterals, promoting blood circuLation to remove blood stasis make contribute to relieve the symptom of Qi-Yin defieiency.Creative Points:According to Qi-Yin defieiency, to explored into the effect and mechanism of invigorating Qi and nourishing Yin, distributing fluid and free collaterals, promoting blood circuLation to remove blood stasis, show that it couLd inhibit secretion of cytokineTNF-a, IL-6, reduce the immune inflammation reaction, protect fonction of saliva, increase AQP5expression, promote saliva secretion, improve symptom of dry, it further illustrated the treatment of Chinese medicine was muLti-channel, muLti-link and muLti-arget.