Dissertation
Dissertation > Medicine, health > Chinese Medicine > Chinese medicine practitioners of other disciplines > TCM neurology and psychiatry

The Zhongjiling Anti myasthenia gravis mechanism

Author WangJing
Tutor ZhouYong
School Beijing University of Traditional Chinese Medicine
Course Chinese and Western medicine combined with the basis
Keywords Zhongjiling Myasthenia gravis EAMG model IFN-γ IL-4 TGF-β Apoptosis
CLC R277.7
Type PhD thesis
Year 2004
Downloads 245
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Myasthenia gravis (Myasthenia Gravis, MG) is a human acetylcholine receptors (Acetylcholine Receptor, N2-AChR) of the neuromuscular junction postsynaptic membrane autoantibody mediated organ-specific autoimmune disease, the main clinical characteristics is involuntary muscle weakness and no sex. Experimental autoimmune myasthenia gravis (Experimental Autoimmune Myasthenia Gravis, EAMG) is extracted from electric rays electric organ acetylcholine receptor of complete Freund's adjuvant (the Complete Freund,'s Adjuvant, CFA) made immune sensitive animals after human severe gravis model. The MG body of N2-AchRab the generation mainly depend on the regulation of Th cells and the secretion of cytokines related to the development of Th1 cytokine IFN-γ, IL-2, and Th2 cytokines IL-4, IL-10 and MG and EAMG. Th3 cytokine TGF-β inhibit the disease. Piece Zhongjiling Hebei Yiling Pharmaceutical Research Institute of Clinical long been used in traditional Chinese medicine for the treatment of myasthenia gravis, a major component of the astragalus, ginseng, deer horn of brucine, with a temperature Ideachip Yang, Fu Yuan Zhen decadent role, clinical the treatment of patients with more than a thousand cases of muscle weakness, the effect is obvious. The pharmacodynamic study suggests Zhongjiling piece has immunomodulatory effects. To investigate the mechanism of immunomodulatory effects Zhongjiling tablet, Lewis rats were immunized with purified N2-AchR, expand research especially cytokines IFN-γ. This topic is divided into four parts: 1. Zhongjiling tablets in the treatment of major pharmacological effects of the administration of EAMG rats from electric rays electric organs extracted containing the N2-AchR crude extract plus complete Freund's adjuvant 150-180gLewis rats made of EAMG, animals at 25 days after the first immunization, administration, clinical symptoms of EAMG rats, muscle strength, exercise capacity, EMG, serum anti-N2 the N2-AchRab, postsynaptic membrane -AchR number of cellular immune function were observed. The results found that heavy muscle Tablets can improve the clinical symptoms of EAMG model rats, improve EAMG model rats body weight, muscle strength and athletic ability, inhibit the EAMG model serum anti-N2-AchRab, reduce EAMG rat model of continuous low-frequency electric stimulation appears after muscle contraction amplitude attenuation, to improve the number of EAMG model rats postsynaptic membrane N2-AchR. Suppression the EAMG model in rats N2-AchR-specific cellular immune responses induced enhancement of cellular immune responses induced by ConA (P lt; 0.01 or P lt; 0.05). Western medicine the pharmacological effects Zhongjiling is different from the immunosuppressive agent with selective immunomodulatory effects that down for the N2-AchR specific pathological autoimmune reaction, but does not inhibit normal immune function, enhanced role. 2 Zhongjiling immune function of normal mice and immune suppression we use 18-22g C57BL/6j female mice orally administered 10 days after discovery, Zhongjiling large, middle dose group of normal mice. spleen, thymus weight had no significant effect. Large doses of Zhongjiling significantly enhanced ConA and LPS-induced mouse spleen T and B lymphocyte proliferation and IL-2 secretion; selection of 18-22g C57BL/6j female mice cyclophosphamide every other day by intraperitoneal injection, dose 80mg/Kg, a total of three times, the preparation of immunosuppressive model results found Zhongjiling can significantly improve the immunosuppressed mice thymus and spleen lt; WP = gt; the 2 Zhongjiling Anti myasthenia gravis mechanism of action studies index, enhanced ConA, LPS-induced T and B cell proliferation, increase the secretion of IL-2, and there is a clear dose-effect relationship. Description Zhongjiling immunostimulants characteristics. 3. N2-AchR Purification and create a purified N2-AchR induced EAMG model containing N2-AchR TritonX-100 crude extract using α-glasses venom agarose affinity chromatography and further purification, the purified N2-AchR after SDS polyacrylamide gel electrophoresis showed four bands, molecular weights were 40,000,43,000,53,000,62,000. This may be the four subunits of the receptor. Coomassie brilliant blue G250 determination of the amount of protein purified N2-AchR 116.6mg/5.2 kg of electric rays wet organ. Reference to foreign literature, N2-purified AchR to of 150 ug domestic electric rays immune Lewis rats every 10 days, a total of three times, successfully induced EAMG model. Purified N2-AchR induced the EAMG model of early onset, severe illness, the model high incidence (gt; 70%). 4. Zhongjiling tablet studies on purified N2-AchR induced of EAMG rats immunomodulatory mechanism by double antibody sandwich ABC-ELISA and RT-PCR method for the purification of N2-AChR induced EAMG rats of IFN-γ, IL- 4, TGF-β content and the level of mRNA transcripts were detected. Research by upregulation EAMG rats in vivo endogenous found heavy Zhongjiling tablets suppressing a transcription factor the TGF-βmRNA the increasing TGF-β protein synthesis, and secrete a large number of TGF-β effect on IFN-γ, IL-4 thereby suppressing the latter two abnormally high levels of factor. A result, anti-N2-AchRab titers decreased, to the role of anti-gravis. Nitrate reductase method and the chemical colorimetric detection purified N2-AChR induced EAMG rats NO and NOS content. Results found that Zhongjiling tablets can inhibit the pathological NOS and NO production, relieve muscle and nerve tissue damage. Purified N2-AChR induced the EAMG rats nest inguinal lymph node cells were cultured in vitro stimulation for 24 hours with the N2-AchR, flow cytometry of different subsets of specific T-cell apoptosis rate. The study found that the specific CD4 T cell apoptosis Zhongjiling tablet can significantly improve recovery EAMG lymphocyte homeostatic state, to achieve the purpose of the anti-gravis. Zhongjiling tablet this to promote apoptosis of autoreactive T cells can not be reversed by IFN-γ. The above results show that EAMG presence in the body of the IFN-γ and IL-4 expression increased abnormal phenomenon, both the role of B cells to stimulate the synthesis of a large number of anti AchRIgG1 and IgG2, this role of autoantibodies in the NMJ at the acetylcholine receptor, resulting neuromuscular transmission disorder and induce the generation of the symptoms of myasthenia gravis. Zhongjiling immunomodulatory effects: inhibition of factor TGF-βmRNA transcription to by upregulating EAMG of endogenous protein synthesis increased TGF-β, a large number of the secretion of TGF-β down abnormal increase in the pathogenic factor --- IFN-γ, IL- 4 results EAMG rat body

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