本文選題：巨噬細(xì)胞 + 淫羊藿��； 參考：《廣州中醫(yī)藥大學(xué)》2016年碩士論文

【摘要】：目的：探討中藥淫羊藿、巴戟天提取物對恒河猴單核衍生的巨噬細(xì)胞在不同極化條件下基因表達(dá)的影響,分析這兩種中藥可能的免疫調(diào)節(jié)的作用機(jī)制。方法：密度梯度離心法分離中國恒河猴外周血單個核細(xì)胞(PBMCs),采用貼壁法分離出單核細(xì)胞,并以粒細(xì)胞巨噬細(xì)胞集落刺激因子(GM-CSF)刺激5天以使單核細(xì)胞分化成巨噬細(xì)胞,隨后不加入極化因子形成MO型巨噬細(xì)胞,以干擾素γ (IFN-γ)或地塞米松繼續(xù)培養(yǎng)2天誘導(dǎo)巨噬細(xì)胞極化為M1和M2c表型,同時分別加入淫羊藿、巴戟天提取物進(jìn)行干預(yù),隨后提取mRNA并逆轉(zhuǎn)錄后,以實時熒光定量PCR檢測CD4、 FoxP3、CTLA-4、CCR5、IDO、IL-10、TGF-β等基因表達(dá)量,以GAPDH為參照基因,與空白組進(jìn)行對比。結(jié)果：與空白對照組相比,加入巴戟天提取物培養(yǎng)的MO巨噬細(xì)胞基因表達(dá)上調(diào)不明顯；經(jīng)淫羊藿提取物培養(yǎng)的MO巨噬細(xì)胞,CCR5基因表達(dá)量上調(diào)4.21倍,TGF-β上調(diào)7.66倍,而CD4、CTLA-4、FoxP3、IL-10、IDO的基因表達(dá)無明顯變化。經(jīng)淫羊藿提取物刺激的M1型巨噬細(xì)胞,IL-10的表達(dá)量上調(diào)11.83倍,FoxP3上調(diào)4.55倍,IDO、CCR5、 CD4、CTLA-4無明顯變化, TGF-β未測出；而由巴戟天提取物刺激的M1型巨噬細(xì)胞,CTLA-4基因表達(dá)量上調(diào)3.22倍,FoxP3上調(diào)3.69倍,CCR5、CD4、IL-10、IDO均無明顯變化,TGF-β未測出。經(jīng)淫羊藿提取物培養(yǎng)刺激的M2c型巨噬細(xì)胞,CCR5基因增加5.94倍,TGF-β基因增加3.53倍,CTLA-4、FoxP3、IDO、IL-10等基因無明顯變化,經(jīng)巴戟天提取物刺激的M2c型巨噬細(xì)胞,CCR5基因增加6.43倍,CTLA-4基因增加3.28倍,IL-10基因增加3.76倍,TGF-β基因增加11.05倍,CD4基因增加3.77倍,FoxP3和IDO兩個基因無明顯變化。結(jié)論：經(jīng)淫羊藿提取物培養(yǎng)的MO型巨噬細(xì)胞,則能夠上調(diào)CCR5和TGF-β的基因表達(dá),呈現(xiàn)出促炎、促纖維化的作用。經(jīng)淫羊藿和巴戟天提取物培養(yǎng)后,M1型巨噬細(xì)胞能上調(diào)IL-10、FoxP3、CTLA-4等抑炎、抑纖維化基因表達(dá),減輕炎癥反應(yīng)和組織纖維化,有利于組織結(jié)構(gòu)的維持和保護(hù)。通過淫羊藿和巴戟天提取物培養(yǎng)后,M2c型巨噬細(xì)胞的CCR5、CD4和TGF-β基因表達(dá)大量增加。表現(xiàn)出促炎和促纖維化的免疫調(diào)節(jié)作用�？偨Y(jié)本實驗的結(jié)果,淫羊藿和巴戟天提取物通過對不同極化條件下的巨噬細(xì)胞基因表達(dá)影響的不同,發(fā)揮促炎和抑炎、促纖維化和抑制纖維化的雙向調(diào)節(jié)作用。
[Abstract]:Aim: to investigate the effect of Herba Epimedii and Morinda officinalis extract on gene expression of monocyte derived macrophages in rhesus monkey under different polarization conditions and to analyze the possible immunomodulatory mechanism of these two herbs. Methods: PBMCs were isolated from the peripheral blood mononuclear cells of Chinese rhesus monkeys by density gradient centrifugation. Monocytes were isolated by adherent method and stimulated with granulocyte macrophage colony stimulating factor (GM-CSF) for 5 days to differentiate monocytes into macrophages. No polarizing factor was added to form MO type macrophages, and the macrophages were further cultured with interferon 緯 (IFN- 緯) or dexamethasone for 2 days to induce the polarization of macrophages into M1 and M2c phenotypes, and the extracts of Herba Epimedii and Morinda officinalis were added for intervention, respectively. Then mRNA was extracted and reverse transcripted, and the expression of CD4, FoxP3 CTLA-4 and CCR5 IDO5 IL-10TGF- 尾 was detected by real-time fluorescence quantitative PCR. GAPDH was used as the reference gene and compared with the control group. Results: compared with the blank control group, the gene expression of MO macrophages cultured with the extract of Morinda officinalis was not significantly up-regulated, and the expression of CCR5 gene was increased by 4.21 times and 7.66 times by TGF- 尾 in the MO macrophages cultured with Herba Epimedii extract. There was no significant change in the expression of IL-10 IDO gene in CD _ 4 ~ + CTLA-4 ~ (- 4) FoxP _ (3) and FoxP _ (3). The expression of IL-10 in M 1 macrophages stimulated by Herba Epimedii extract was increased by 11.83 times and FoxP3 by 4.55 times, while CD4 CTLA-4 did not change, but TGF- 尾 was not detected. However, the expression of CTLA-4 gene in M1 macrophages stimulated by Morinda officinalis extract increased 3.22 times and FoxP3 increased 3.69 times. The CCR5 gene of M 2c macrophages stimulated by Herba Epimedii extract increased 5.94 times and the TGF- 尾 gene increased 3.53 times. The increase of CCR5 gene in M 2c macrophages stimulated by Morinda officinalis extract was 6.43 times. CTLA-4 gene increased 3.28 times and IL-10 gene increased 3.76 times. TGF- 尾 gene increased 11.05 times and CD4 gene increased 3.77 times. There was no significant change in two genes: FoxP3 and IDO. Conclusion: the MO type macrophages cultured with Herba Epimedii extract can up-regulate the gene expression of CCR5 and TGF- 尾, which can promote inflammation and fibrosis. After culture of Herba Epimedii and Morinda officinalis extracts, M 1 macrophages could up-regulate the expression of anti-inflammatory genes, such as IL-10, FoxP3, CTLA-4, reduce inflammatory reaction and tissue fibrosis, and help to maintain and protect the tissue structure. After cultured with Herba Epimedii and Morinda officinalis extract, the expression of CCR5, CD4 and TGF- 尾 genes in M2c macrophages was significantly increased. It showed immunomodulatory effect of promoting inflammation and fibrosis. The results showed that the extracts of Herba Epimedii and Morinda officinalis had different effects on the gene expression of macrophages under different polarization conditions, and played a bidirectional role in promoting inflammation and inhibiting inflammation, promoting fibrosis and inhibiting fibrosis.
【學(xué)位授予單位】：廣州中醫(yī)藥大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R285

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 胡蓉;陳漢銳;許錚弟;林麗珠;;喘可治注射液聯(lián)合化療對老年非小細(xì)胞肺癌患者生存質(zhì)量的影響[J];廣州中醫(yī)藥大學(xué)學(xué)報;2015年05期

2 馮毅;;喘可治注射液對慢性阻塞性肺疾病大鼠血清炎癥因子及肺功能的影響[J];中醫(yī)學(xué)報;2015年09期

3 喻隼鋒;高山;趙蘇;;喘可治注射液對支氣管哮喘急性發(fā)作期患者外周血白細(xì)胞介素-17水平的影響[J];醫(yī)藥導(dǎo)報;2015年09期

4 傅淑平;楊麗;洪浩;偶晨;張榮華;;淫羊藿苷促SD大鼠骨髓間充質(zhì)干細(xì)胞骨向分化作用的實驗研究[J];中國中西醫(yī)結(jié)合雜志;2015年07期

5 李丹青;肖強(qiáng)兵;;巴戟天多糖通過p38 MAPK信號通路促間充質(zhì)干細(xì)胞成骨分化的體外研究[J];中國中醫(yī)骨傷科雜志;2015年05期

6 張會英;邢博;王晉榮;王平;;喘可治注射液治療小兒咳嗽變異性哮喘療效觀察[J];臨床合理用藥雜志;2015年13期

7 黃純美;張蓓;許仕杰;李耿;單麗囡;林旋齡;劉小虹;;喘可治注射液對COPD大鼠IL-12/IL-12R/STAT4信號通路的影響[J];中藥新藥與臨床藥理;2015年01期

8 陳漢銳;胡蓉;曹洋;黃學(xué)武;黎鵬;許錚弟;林麗珠;;喘可治注射液聯(lián)合化療治療老年非小細(xì)胞肺癌近期療效觀察[J];中藥新藥與臨床藥理;2014年05期

9 陳瀅宇;陳頌;符林春;郭衛(wèi)中;鄧文娣;賴春輝;周映云;于靚;;中藥注射液喘可治對艾滋病猴模型Th17和Treg平衡的影響[J];廣州中醫(yī)藥大學(xué)學(xué)報;2014年04期

10 黃東暉;王慧賢;吳少麗;黎潭輝;;喘可治注射液對小鼠氣道炎癥Th17與Treg的影響[J];北京中醫(yī)藥大學(xué)學(xué)報;2014年05期

相關(guān)會議論文 前1條

1 盧耀增;吳小閑;符林春;郭衛(wèi)中;陳頌;鄧文娣;羅紅梅;周映云;賴春輝;杜式群;;“喘可治”治療SIV/SAIDS猴的療效觀察[A];中華中醫(yī)藥學(xué)會防治艾滋病學(xué)術(shù)研討會暨2006年年會論文集[C];2006年

相關(guān)博士學(xué)位論文 前3條

1 陳瀅宇;喘可治對艾滋病猴免疫調(diào)節(jié)作用及Th17分化的機(jī)制研究[D];廣州中醫(yī)藥大學(xué);2014年

2 黃純美;中藥喘可治注射液對COPD大鼠IL-12/STAT4、IL-4/STAT6信號通路影響研究[D];廣州中醫(yī)藥大學(xué);2014年

3 曾麗絢;淫羊藿苷抑制大鼠慢性阻塞性肺疾病炎癥的實驗研究[D];復(fù)旦大學(xué);2010年

相關(guān)碩士學(xué)位論文 前5條

1 朱可;喘可治對非小細(xì)胞肺癌免疫功能及骨髓抑制影響的臨床研究[D];廣州中醫(yī)藥大學(xué);2014年

2 黃文通;喘可治注射液輔助治療慢性阻塞性肺疾病急性加重期臨床療效觀察[D];北京中醫(yī)藥大學(xué);2013年

3 胡蓉;喘可治注射液聯(lián)合化療治療老年非小細(xì)胞肺癌的臨床研究[D];廣州中醫(yī)藥大學(xué);2013年

4 陳文喜;喘可治注射液對慢性阻塞性肺疾病患者外周血單個核細(xì)胞炎癥介質(zhì)的影響[D];福建中醫(yī)藥大學(xué);2012年

5 劉文華;喘可治注射液對支氣管哮喘免疫調(diào)控作用的臨床與實驗研究[D];廣州中醫(yī)藥大學(xué);2011年

，

本文編號：1966369