本文關(guān)鍵詞： 姜黃素 HaCaT細(xì)胞 氮酮 凝膠 銀屑病 小鼠 gamma/delta T細(xì)胞 細(xì)胞因子　出處：《第二軍醫(yī)大學(xué)》2013年博士論文　論文類型：學(xué)位論文

【摘要】：姜黃素是從姜科植物姜黃(Curcuma longa)中提取的一種酚類化合物,是植物界很稀少的具有二酮的色素。姜黃素能夠通過抑制多種絲氨酸/蘇氨酸蛋白激酶的磷酸化,如蛋白激酶A、蛋白激酶C、精蛋白激酶、磷酸化酶激酶、自身磷酸化活性蛋白激酶以及pp60c-src酪氨酸激酶等,影響細(xì)胞的信號轉(zhuǎn)導(dǎo),表現(xiàn)出多樣化的生物學(xué)作用,包括影響轉(zhuǎn)錄因子、酶、細(xì)胞因子和生長因子等,從而展現(xiàn)多重藥理活性,如促進(jìn)凋亡,抗增生,抗氧化,抗炎,抗腫瘤和抗血管增生等作用。 姜黃素不溶于水,可應(yīng)用的靜脈給藥方法至今未見報道。國內(nèi)外目前已有的臨床研究資料都是以口服給藥為途徑。但是姜黃素的生物利用度之低卻另人失望。在一項姜黃素Ⅰ期臨床研究中,口服3.6g/天的劑量也僅僅能夠產(chǎn)生11.1±0.6nmol/L的血漿峰濃度,而低于此劑量則未檢測到姜黃素。鑒于以上原因,雖然眾多關(guān)于姜黃素的體內(nèi)外研究已明確其藥理作用,但是它極低的生物利用度嚴(yán)重制約了對其應(yīng)用研究的腳步。 銀屑病是臨床常見的皮膚病,發(fā)病率占世界人口的0.1%-3%。因該病頑固難治,仍被列為當(dāng)今世界皮膚科領(lǐng)域的重要研究課題,是全世界皮膚科重點防治疾病之一。雖然銀屑病的發(fā)病機理目前尚未完全明了,但越來越多的證據(jù)已經(jīng)證明IL-23→IL-17→IL-22作用鏈以及其他細(xì)胞因子在疾病的發(fā)展中起了至關(guān)重要的作用。銀屑病皮損區(qū)皮膚在基因與蛋白水平均可檢測到IL-23, IL-21，IL-22和IL-17的高表達(dá),并且IL-23可在小鼠皮膚誘發(fā)與人類銀屑病類似的病變特征,如紅斑,表皮增生(棘皮征),角化不全和白細(xì)胞浸潤,同時真皮層的炎癥與棘皮征被認(rèn)為與細(xì)胞因子IL-22有關(guān),此過程還需趨化因子受體CCR6的參與。IL-23誘導(dǎo)的皮膚炎癥主要通過IL-17及其下游相關(guān)細(xì)胞因子實現(xiàn)的。除了上述提及的細(xì)胞因子以外,尚有TNF-α, IL-lα, IL-1β, IL-6, IL-8, VEGF, TGF-β等參與銀屑病的病理生理過程。 銀屑病參與的細(xì)胞因子眾多,牽涉到的信號轉(zhuǎn)導(dǎo)更是十分龐雜�，F(xiàn)在已知的涉及到NF-κB激活的細(xì)胞因子有IL-17A, IL-17F, TNF-α, IL-6等。姜黃素多靶點的作用機制與銀屑病復(fù)雜的信號轉(zhuǎn)導(dǎo)網(wǎng)絡(luò)有諸多的疊加之處,如抑制NF-κB激活、抑制STAT3磷酸化等�？紤]到姜黃素在銀屑病治療中潛在的應(yīng)用價值,著眼于銀屑病病變部位的淺表特點,本研究通過驗證姜黃素對其靶細(xì)胞——KC細(xì)胞的藥理作用基礎(chǔ)上,通過局部給藥的方式(規(guī)避姜黃素口服給藥難以在銀屑病靶細(xì)胞產(chǎn)生有效濃度的難題),通過小鼠銀屑病動物模型評估姜黃素局部給藥的治療效果,為后續(xù)的應(yīng)用研究打下堅實的實驗基礎(chǔ)。 我們的研究發(fā)現(xiàn)：TRAIL及其受體在姜黃素誘導(dǎo)的HaCaT細(xì)胞凋亡中起重要作用,此作用包括TRAIL-R1和TRAIL-R2表達(dá)的上調(diào)以及凋亡抑制蛋白BcL-XL, IAP-1,IAP-2表達(dá)的抑制。以上證據(jù)說明TRAIL及其受體通路在姜黃素誘導(dǎo)細(xì)胞凋亡的作用中占有重要地位。此外姜黃素還能夠抑制TNF-α誘導(dǎo)的NF-κB的激活,并減少炎癥因子IL-6和IL-8的分泌。這無疑使其具備了理想的,對抗銀屑病中TNF-α作用的潛力。借鑒以往的研究基礎(chǔ),在原有配方的基礎(chǔ)上,引入了另外的透皮促進(jìn)劑：氮酮,并適當(dāng)調(diào)整了組方的比例,成功配制出了符合實驗研究需要、穩(wěn)定、能夠強效促透的姜黃素凝膠制劑。以此制劑為基礎(chǔ),我們驗證評估了姜黃素對咪喹莫特誘導(dǎo)的銀屑病小鼠模型的實驗治療效果,結(jié)果令人振奮。咪喹莫特小鼠銀屑病模型最早于2009年見諸報道,是迄今為止最能夠最大程度模擬銀屑病發(fā)病機理的模型。我們發(fā)現(xiàn)姜黃素可以抑制咪喹莫特導(dǎo)致的皮膚增厚和炎癥反應(yīng)；顯著抑制咪喹莫特誘導(dǎo)的銀屑病模型小鼠耳廓皮膚的IL-17A, IL-17F, IL-22, IL-1β, TNF-α和IL-6的mRNA水平的上升。并且還發(fā)現(xiàn),姜黃素可能會抑制咪喹莫特誘導(dǎo)的實驗小鼠真皮丫δ T細(xì)胞的增殖,而這一細(xì)胞被認(rèn)為在銀屑病的發(fā)病機制中占有重要地位。 本研究在細(xì)胞和實驗動物水平上系統(tǒng)的探討了姜黃素對于銀屑病治療的機理和效果評估,并為實現(xiàn)這一研究目標(biāo)對姜黃素的外用凝膠制劑進(jìn)行了研究和改進(jìn)。首次在銀屑病動物模型上觀察研究了姜黃素對IL-23/IL-17A/IL-22細(xì)胞因子軸的影響,并和類固醇激素的作用進(jìn)行了對比,為該化合物未來的開發(fā)方向奠定了重要的理論和實驗基礎(chǔ)。
[Abstract]:Curcumin is from Zingiberaceae turmeric (Curcuma longa) is a kind of phenolic compounds extracted from plants, is very rare with two ketone pigment. Curcumin can inhibit a variety of the serine / threonine protein kinase phosphorylation, such as protein kinase A, protein kinase C, egg white kinase and phosphorylase kinase itself. The phosphorylation activity of protein kinase pp60c-src and tyrosine kinase, signal transduction of cells, showing diverse biological effects, including the effects of transcription factors, enzymes, cytokines and growth factors, so as to show a variety of pharmacological activities, such as promoting apoptosis, anti proliferative, antioxidant, anti-inflammatory, anti-tumor and anti angiogenesis effect.
Curcumin is not soluble in water, can be used for intravenous administration method has not been reported at home and abroad. The current clinical research data are based on the existing oral administration way. But the low bioavailability of curcumin was disappointing. In a phase I clinical study of curcumin, peak plasma concentration of oral 3.6g/ days dose only can produce 11.1 + 0.6nmol/L, and lower than the dose not detected by curcumin. In view of the above reasons, although a number of curcumin in vitro and in vivo studies have confirmed its pharmacological effects, but its low bioavailability seriously restrict the application of the pace.
Psoriasis is a common skin disease in clinic, the incidence rate of 0.1%-3%. of the world's population with the disease is difficult to cure, still listed as an important research topic in the world in the field of Department of Dermatology, Department of dermatology is one of the world's focus on disease prevention and control. Although the pathogenesis of psoriasis is not yet entirely clear, but more and more evidences have shown that IL-23, IL-17 IL-22, chain and other cytokines play a crucial role in the development of the disease. The skin lesions of patients with psoriasis in gene and protein level were detected in IL-23, IL-21, high expression of IL-22 and IL-17, and IL-23 in the pathological characteristics, and human induced psoriasis like mouse skin such as erythema, epidermal hyperplasia (spine skin syndrome), parakeratosis and leukocyte infiltration and inflammation and dermal acanthosis syndrome were found and cytokines related to IL-22, this process also need to chemokine receptor CCR6 Skin inflammation induced by.IL-23 is mainly achieved by IL-17 and its downstream related cytokines. Besides the above mentioned cytokines, there are TNF-, IL-l, IL-1, IL-6, IL-8, VEGF and TGF- beta participate in the pathophysiology process of psoriasis.
Cytokines in psoriasis in many signal transduction involved is very complex. Now known to involve NF- cell factor kappa B activation of IL-17A, IL-17F, TNF- alpha, IL-6. Signal transduction mechanism and psoriasis curcumin targets have many complex superimposed place, such as inhibition of NF- k the activation of B, inhibit the phosphorylation of STAT3. Considering the potential application value of curcumin in the treatment of psoriasis, superficial characteristics focus on psoriasis lesion, this study verified by curcumin in KC cells and the target cells of pharmacological basis, through local administration way (oral administration is difficult to avoid the problem of curcumin in the target cell of psoriasis produce effective concentration), topical therapy through mice psoriasis animal model for experimental evaluation of curcumin, a solid foundation for the application of subsequent lay.
Our study found that: TRAIL and its receptors play an important role in HaCaT cell apoptosis induced by curcumin in this role, including the expression of TRAIL-R1 and upregulation of TRAIL-R2 and inhibitor of apoptosis protein BcL-XL, IAP-1, inhibit the expression of IAP-2. The above evidence plays an important role of TRAIL and its receptor pathway in curcumin induced apoptosis. The activation of the addition of curcumin TNF- can also inhibit NF- induced by alpha kappa B, and reduce the secretion of inflammatory cytokines IL-6 and IL-8. This undoubtedly makes it have the ideal effect against psoriasis TNF- alpha potential. Basis of previous studies, based on the original formula, the penetration enhancers azone, and another. Appropriate adjustment of the prescription ratio was prepared successfully with the experimental research, stability, can be powerful penetration. This gel preparation of curcumin based formulations, we verify the evaluation Evaluate the experimental therapeutic effect of curcumin on psoriasis animal model induced by imiquimod, exciting results. Imiquimod psoriasis model mice. The earliest reported in 2009, is by far the most to the greatest extent simulating the pathogenesis of psoriasis model. We found that skin thickening and inflammatory reaction of curcumin can inhibit the cause of imiquimod; significant inhibition of imiquimod induced psoriasis model mice auricle skin IL-17A, IL-17F, IL-22, IL-1 beta, TNF- alpha and IL-6 increased the level of mRNA. It was also found that curcumin may inhibit imiquimod induced mice dermal Ya delta proliferation of T cells, and the cells are thought to play an important role in the pathogenesis of psoriasis.
In this study, experimental animal cells and on the level of the system is discussed for the mechanism and effect of curcumin in the treatment of psoriasis and evaluation, research and improvement for the realization of the goal of topical gel preparation of curcumin was carried out. For the first time in animal models of psoriasis were studied on the effects of curcumin on IL-23/IL-17A/IL-22 cell factor axis, and comparison of the effects of steroid hormones, laid an important theoretical and experimental basis for the development direction of the compound in the future.

【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2013
【分類號】：R758.63;R-332

【共引文獻(xiàn)】

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相關(guān)會議論文 前3條

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相關(guān)博士學(xué)位論文 前10條

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7 劉軍麟;PDCD1基因與系統(tǒng)性紅斑狼瘡以及維生素D受體基因與銀屑病關(guān)聯(lián)研究的Meta-分析[D];安徽醫(yī)科大學(xué);2008年

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相關(guān)碩士學(xué)位論文 前10條

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8 陳小娥;EGCG對急性UVB輻射后表皮角質(zhì)形成細(xì)胞的保護(hù)機制探討及對慢性UVA輻射后小鼠皮膚光老化的防護(hù)作用[D];南京醫(yī)科大學(xué);2008年

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10 任為;IL23R基因多態(tài)性與漢族人銀屑病易感性關(guān)聯(lián)分析研究[D];安徽醫(yī)科大學(xué);2009年

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