本文選題：熊果酸　切入點(diǎn)：外周血淋巴細(xì)胞　出處：《武漢大學(xué)》2005年碩士論文

【摘要】：背景及目的: 熊果酸為五環(huán)三萜類化合物,廣泛存在于中草藥和食品中,具有廣泛的生物學(xué)活性和藥理學(xué)功效,如抗腫瘤、抗炎、免疫調(diào)節(jié)和恢復(fù)造血等。在熊果酸對(duì)機(jī)體免疫功能影響的研究中,You等發(fā)現(xiàn)熊果酸可通過(guò)促進(jìn)巨噬細(xì)胞釋放TNF-α和NO從而增強(qiáng)細(xì)胞免疫功能。輔助性淋巴細(xì)胞(Th細(xì)胞)在機(jī)體的免疫中起著極為關(guān)鍵的作用,尤其是通過(guò)分泌細(xì)胞因子調(diào)節(jié)免疫功能,目前尚無(wú)熊果酸對(duì)Th細(xì)胞影響的報(bào)道。本實(shí)驗(yàn)旨在研究熊果酸對(duì)正常人外周血淋巴細(xì)胞分泌Th1型細(xì)胞因子(IFN-γ,和IL-2)的影響,并檢測(cè)Th1型細(xì)胞因子分泌過(guò)程中MAPK的變化,對(duì)機(jī)制作初步探討。 方法: 采用密度梯度離心法分離正常人外周血單個(gè)核細(xì)胞,再用黏附法分離得到外周血淋巴細(xì)胞。實(shí)驗(yàn)分別用不同濃度熊果酸(5～30μM)及PHA/PMA刺激外周血淋巴細(xì)胞,采用MTT實(shí)驗(yàn)分析刺激24小時(shí)的淋巴細(xì)胞的增殖情況,雙抗體夾心法ELISA和半定量RT-PCR分別檢測(cè)Th1型細(xì)胞因子(IFN-γ和IL-2)分泌及mRNA表達(dá)的變化;Western Blot檢測(cè)p38 MAPK的表達(dá)及活化水平。 結(jié)果: 淋巴細(xì)胞在不同濃度熊果酸作用后,各組細(xì)胞增殖無(wú)顯著差異,而Th1型細(xì)胞因子(IFN-γ和IL-2)的分泌和mRNA的表達(dá)水平均隨熊果酸濃度的增加呈濃度依賴性下降(P0.05),在高濃度熊果酸作用后,p38MAPK的活化水平出現(xiàn)顯著性降低。 結(jié)論: 熊果酸在本研究濃度范圍內(nèi)對(duì)絲裂原活化的淋巴細(xì)胞增殖無(wú)影響,但顯著抑制Th1型細(xì)胞因子的分泌和mRNA的表達(dá),提示抑制可能是在轉(zhuǎn)錄和/或轉(zhuǎn)錄后水平發(fā)揮作用,抑制p38MAPK的活化可能是其機(jī)制之一。熊果酸和地塞米松分子結(jié)構(gòu)相似,推測(cè)熊果酸可能通過(guò)類似地塞米松下調(diào)糖皮質(zhì)激素受體發(fā)揮作用。熊果酸有可能用于自身免疫性疾病和超敏反應(yīng)性疾病的防治。
[Abstract]:Background and purpose:
Ursolic acid is three pentacyclic triterpenoids, widely exists in Chinese herbal medicine and food, has a wide range of biological activities and pharmacological effects, such as anti-tumor, anti-inflammatory, immune regulation and hematopoietic recovery. Study the effects of ursolic acid on immune function, You found that ursolic acid can stimulate macrophages to release TNF- alpha and NO can enhance the cellular immune function. Helper lymphocytes (Th cells) in immune plays a key role, especially through the secretion of cytokines regulate the immune function, there is no reported effects of ursolic acid on Th cells. This experiment aims to study on ursolic acid in normal human peripheral blood lymphocyte Th1 cytokines (IFN-, and IL-2) the influence, and to detect changes of Th1 cytokine secretion process in MAPK, preliminary study on the mechanism.
Method:
Using density gradient centrifugation of normal human peripheral blood mononuclear cells isolated from peripheral blood lymphocytes by adhesion method. Experiments with different concentration of ursolic acid (5~30 M) and PHA/PMA stimulated peripheral blood lymphocytes, the analysis of the proliferation of lymph cells stimulated 24 hours MTT experiment, double antibody sandwich ELISA and semi quantitative RT-PCR were used to detect Th1 type cytokines (IFN- gamma and IL-2) changes in the secretion and expression of mRNA; expression of Western Blot p38 MAPK detection and activation level.
Result:
Lymphocytes in different concentration of ursolic acid. After that, there was no significant difference between the cells proliferation, and Th1 type cytokines (IFN- gamma and IL-2) expression and mRNA secretion were increased with the increase of the concentration of ursolic acid in a concentration dependent decrease (P0.05), the high concentration of ursolic acid after activation level of p38MAPK significantly decreased.
Conclusion:
No effect of ursolic acid on the activation of mitogen in this study in the concentration range of lymphocyte proliferation, secretion and expression of mRNA but inhibited Th1 type cytokines, suggesting that inhibition may be in transcriptional and / or post transcriptional levels play a role in inhibiting the activation of p38MAPK may be one of the mechanisms of ursolic acid and dexamethasone. Similar molecular structure. That ursolic acid may play a role through the down-regulation of glucocorticoid receptor is similar to dexamethasone. Ursolic acid may be used for the prevention and treatment of autoimmune diseases and allergic diseases.

【學(xué)位授予單位】：武漢大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2005
【分類號(hào)】：R392

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