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雷公藤多苷上調(diào)克羅恩病病人局部腸粘膜Treg細胞及調(diào)節(jié)炎癥因子的分泌

發(fā)布時間:2018-05-30 02:22

  本文選題:克羅恩病 + 雷公藤多苷。 參考:《南京大學(xué)》2014年碩士論文


【摘要】:背景:克羅恩病(Crohn's disease, CD)屬于炎癥性腸病(inflammatory bowel disease,IBD)的一種,近一個世紀以來其發(fā)病率越來越高。盡管相關(guān)研究逐漸深入,但其確切病因至今仍然不清。目前認為其病因涉及基因易感性、環(huán)境因素、免疫因素。表達叉狀頭/翅膀狀螺旋轉(zhuǎn)錄因子(Forkhead Box P3, Foxp3)的調(diào)節(jié)性T細胞(t regulatory cells, Tregs)通過分泌IL-10、TGF-β發(fā)揮抑炎作用,在維持機體免疫穩(wěn)態(tài)方面發(fā)揮重要作用。許多研究發(fā)現(xiàn)Tregs與CD發(fā)病之間存在相關(guān)性。雷公藤多苷提取自中國一味傳統(tǒng)中藥雷公藤的根部,具有抗炎、免疫調(diào)節(jié)、抗腫瘤等作用。在中國,雷公藤多苷被廣泛用于治療多種自身免疫性疾病,包括類風(fēng)濕關(guān)節(jié)炎、腎炎及系統(tǒng)性紅斑狼瘡等。雷公藤能夠緩解結(jié)腸炎小鼠的腸道炎癥,其作用機制包括抑制TLRs/NF-κB和TNF-α/TNFR2信號通路,抑制L-6/STAT3通路、下調(diào)IL-17表達。我們之前研究證實,雷公藤多苷能夠有效誘導(dǎo)CD緩解。但其具體機制尚未明確。目的: 探索雷公藤多苷對CD病人腸道粘膜局部炎癥因子表達及Foxp3+ Tregs水平的作用。方法:從南京軍區(qū)南京總醫(yī)院門診部入選處于中重度活動期的CD病人,給予口服雷公藤多苷片60mg/天(中國江蘇,泰州制藥廠)。入選時及三個月后行結(jié)腸鏡檢查,于病變明顯的同一段腸管鉗取6塊腸粘膜。HE染色后使用光鏡觀察腸粘膜病理變化并進行炎癥浸潤程度的評分;采用免疫組化法檢測病變腸粘膜中Foxp3、IL-10、TNF-α的表達變化;采用免疫印跡(Western blot)檢測Foxp3蛋白表達的變化;采用酶聯(lián)免疫吸附法(ELISA)測定病變腸道粘膜中炎癥因子IL-10、TNF-α水平變化。結(jié)果: 20個CD病人參與了本項研究,年齡介于25-57歲之間。其中8個病人疾病處于輕度活動期,另外12個病人疾病處于中度活動期。所有人均接受了三個月的口服雷公藤多苷片治療,沒有人因為疾病不良反應(yīng)而退出實驗。治療前的病變腸粘膜HE病理可見潰瘍、上皮損害、腺體減少和炎癥細胞的浸潤,而接受三個月雷公藤多苷片治療后,鏡下病理明顯改善。治療后的炎癥浸潤程度評分顯著降低(8.1±2.2 vs 6.8±2.6,P0.05)。免疫組化的結(jié)果表明,雷公藤治療后,腸粘膜中Foxp3+ Tregs和IL-10陽性細胞的水平呈現(xiàn)增多趨勢,而促炎因子TNF-α則呈現(xiàn)下調(diào)趨勢。Western blot檢測顯示,治療前病變腸粘膜中Foxp3蛋白有一定的表達,而雷公藤多苷治療后,Foxp3蛋白能夠進一步上調(diào)。在炎癥因子表達方面,雷公藤多苷使促炎因子TNF-α水平顯著下降(189.6±17.4 vs173.0±19.0,P0.01),而抑炎因子IL-10的水平顯著上升(45.7±16.3 vs 55.6±14.8,P0.05)。結(jié)論:雷公藤多苷能夠緩解CD病變腸管的炎癥,改善局部粘膜的病理損害,調(diào)節(jié)局部免疫環(huán)境的炎癥因子的表達水平,使促炎因子TNF-1α水平下降,抑炎因子IL-10的水平上升,其作用機制可能和病變腸管中Foxp3+ Tregs增多有關(guān)。
[Abstract]:Background: Crohn's disease (CDD) is a kind of inflammatory bowel disease. The exact cause of the disease is still unclear, although the relevant studies have gradually deepened. It is believed that the etiology is related to genetic susceptibility, environmental factors and immune factors. Regulatory T cell regulatory cells, Tregs), which expresses Forkhead Box P3 (Foxp3), plays an important role in maintaining immune homeostasis by secreting IL-10 TGF- 尾. Many studies have found a correlation between Tregs and CD. Tripterygium wilfordii polyglycosides extracted from the roots of traditional Chinese medicine Tripterygium wilfordii have anti-inflammatory, immunomodulatory and anti-tumor effects. In China, tripterygium wilfordii polyglycosides are widely used in the treatment of many autoimmune diseases, including rheumatoid arthritis, nephritis and systemic lupus erythematosus. Tripterygium wilfordii can relieve intestinal inflammation in mice with colitis by inhibiting TLRs / NF- 魏 B and TNF- 偽 / TNFR2 signaling pathway, inhibiting L-6/STAT3 pathway and down-regulating IL-17 expression. Our previous studies have confirmed that triptolide can effectively induce CD remission. But its specific mechanism is not clear. Objective: to investigate the effect of tripterygium wilfordii polyglycoside on the expression of local inflammatory factors and the level of Foxp3 Tregs in intestinal mucosa of CD patients. Methods: CD patients were selected from the outpatient department of Nanjing General Hospital of Nanjing military region and given oral Tripterygium wilfordii polyglycoside tablets for 60mg/ days (Taizhou Pharmaceutical Factory, Jiangsu, China). Colonoscopy was performed at the time of selection and after 3 months. After 6 intestinal mucosa was stained with HE, the pathological changes of intestinal mucosa were observed with light microscope and the degree of inflammatory infiltration was evaluated. The expression of Foxp3hIL-10 TNF- 偽 was detected by immunohistochemistry, the expression of Foxp3 protein was detected by Western blot, and the level of IL-10TNF- 偽 in intestinal mucosa was detected by enzyme-linked immunosorbent assay (Elisa). Results: twenty CD patients, aged 25-57, participated in the study. Among them, 8 patients had mild active disease and 12 patients had moderate active disease. All were treated with oral tripterygium wilfordii for three months, and no one dropped out because of adverse effects. The pathological changes of intestinal mucosa before treatment were ulcer, epithelial damage, decreased glands and infiltration of inflammatory cells. After three months of treatment with tripterygium wilfordii polyglycosides, the pathological changes were obviously improved under microscope. After treatment, the score of inflammatory infiltration decreased significantly (8.1 鹵2.2 vs 6.8 鹵2.6 P 0.05). The results of immunohistochemistry showed that the levels of Foxp3 Tregs and IL-10 positive cells in intestinal mucosa increased after Tripterygium wilfordii treatment, while the levels of TNF- 偽 were down-regulated. Western blot showed that TNF- 偽 was down-regulated. Foxp3 protein was expressed in intestinal mucosa before treatment, but Foxp3 protein was upregulated further after triptolide was treated with tripterygium wilfordii. In terms of inflammatory factor expression, tripterygium wilfordii polyglycoside significantly decreased the level of proinflammatory factor TNF- 偽 by 189.6 鹵17.4 vs173.0 鹵19.0U P0.01g, while the level of IL-10 increased significantly by 45.7 鹵16.3 vs 55.6 鹵14.8P0.05a. Conclusion: Tripterygium wilfordii polyglycoside can alleviate the inflammation of intestinal canal of CD lesion, improve the pathological damage of local mucosa, regulate the expression of inflammatory factor in local immune environment, decrease the level of proinflammatory factor TNF-1 偽 and increase the level of IL-10. The mechanism may be related to the increase of Foxp3 Tregs in the diseased intestine.
【學(xué)位授予單位】:南京大學(xué)
【學(xué)位級別】:碩士
【學(xué)位授予年份】:2014
【分類號】:R574.62

【參考文獻】

相關(guān)期刊論文 前4條

1 ;Triptolide suppresses IL-1β-induced chemokine and stromelysin-1 gene expression in human colonic subepithelial myofibroblasts[J];Acta Pharmacologica Sinica;2007年01期

2 龔劍峰;鈕凌穎;魏曉為;朱維銘;李寧;黎介壽;;腸內(nèi)營養(yǎng)聯(lián)合雷公藤多甙誘導(dǎo)克羅恩病緩解的研究[J];中華外科雜志;2009年16期

3 廖南生;任建安;范朝剛;王革非;趙允召;黎介壽;;雷公藤多甙預(yù)防克羅恩病術(shù)后復(fù)發(fā)[J];中華胃腸外科雜志;2009年02期

4 陶慶松;任建安;嵇振嶺;李俊生;王新波;蔣小華;;雷公藤多甙在維持術(shù)后克羅恩病臨床緩解中的作用[J];中華胃腸外科雜志;2009年05期

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