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依折麥布聯(lián)合羅格列酮通過LXRα-ABCA1途徑對血管平滑肌源性泡沫細胞膽固醇含量的影響

發(fā)布時間:2019-05-27 14:22
【摘要】:目的:研究依折麥布聯(lián)合羅格列酮對氧化低密度脂蛋白(ox-LDL)誘導(dǎo)的血管平滑肌源性泡沫細胞膽固醇含量的影響,探討肝X受體α(LXRα)-三磷酸腺苷結(jié)合盒轉(zhuǎn)運體A1(ABCA1)信號通路在其中的作用。方法:正常培養(yǎng)的原代大鼠血管平滑肌細胞(VSMCs)作為空白對照組,利用ox-LDL(50ug/ml)干預(yù)VSMCs建立平滑肌源性泡沫細胞模型,然后根據(jù)實驗要求將泡沫細胞分為泡沫細胞組、不同濃度的依折麥布組(3umol/L,10umol/L,30umol/L)、羅格列酮組(25umol/L)、聯(lián)合組(30umol/L依折麥布+25 umol/L羅格列酮)。油紅O染色鑒定泡沫細胞模型,real-time PCR檢測各組細胞LXRα和ABCA1的m RNA表達,Western blot測定各組細胞LXRα和ABCA1蛋白表達,酶熒光法檢測各組細胞內(nèi)總膽固醇(TC)及游離膽固醇(FC)的含量。結(jié)果:1.與空白對照組相比,泡沫細胞組LXRα和ABCA1的m RNA表達降低(P0.05);與泡沫細胞組相比,不同濃度的依折麥布組LXRα和ABCA1的m RNA表達增加(P0.05),且呈一定濃度依賴性(P0.05)。2.與空白對照組相比,泡沫細胞組LXRα、ABCA1的m RNA和蛋白表達均降低(P0.05);與泡沫細胞組相比,依折麥布30umol/L組、羅格列酮組及聯(lián)合組LXRα、ABCA1的m RNA和蛋白表達增加(P0.05),且聯(lián)合組與依折麥布30umol/L組和羅格列酮組相比,LXRα、ABCA1的m RNA和蛋白表達增加更為顯著(P0.05)。3.與空白對照組相比,泡沫細胞組細胞內(nèi)TC、FC和膽固醇酯(CE)的含量均增加(P0.05),且CE/TC50%;與泡沫細胞組相比,依折麥布30umol/L組、羅格列酮組及聯(lián)合組細胞內(nèi)TC、FC、CE含量均減少,CE/TC比值下降(P0.05),且聯(lián)合組與依折麥布30umol/L組和羅格列酮組相比,細胞內(nèi)TC、FC、CE含量及CE/TC比值降低更為顯著(P0.05)。結(jié)論:依折麥布及羅格列酮均可以通過上調(diào)LXRα-ABCA1信號通路,促進平滑肌源性泡沫細胞中ABCA1介導(dǎo)的膽固醇逆轉(zhuǎn)運(RCT),減少細胞內(nèi)蓄積的膽固醇;且兩者聯(lián)合可進一步上調(diào)LXRα和ABCA1的表達,促進膽固醇外流,顯著降低細胞內(nèi)膽固醇的含量。
[Abstract]:Objective: to study the effect of efolib combined with rosiglitazone on cholesterol content in vascular smooth muscle foam cells induced by oxidized low density lipoprotein (ox-LDL). To investigate the role of liver X receptor 偽 (LXR 偽)-adenosine triphosphate binding cassette transporter A1 (ABCA1) signaling pathway. Methods: the normal cultured primary rat vascular smooth muscle cells (VSMCs) were used as the blank control group. The smooth muscle foam cell model was established by the intervention of ox-LDL (50ug/ml) in VSMCs. According to the requirements of the experiment, foam cells were divided into foam cell group, different concentrations of ezumabe group (3umol group, 10umol group, 30umol group), rogaridone group (25umol/L), combined group (30umol/L etimab 25umol/L rosiglitazone). The foam cells were divided into foam cell group, different concentrations of ezumabe group (3umol group, 10umol group, 30umol group). The foam cell model was identified by oil red O staining. The expression of LXR 偽 and ABCA1 m RNA in each group was detected by real-time PCR. The expression of LXR 偽 and ABCA1 protein in each group was detected by, Western blot. The contents of total cholesterol (TC) and free cholesterol (FC) in each group were detected by enzyme fluorescence method. Result: 1. Compared with the blank control group, the expression of m RNA of LXR 偽 and ABCA1 in foam cell group was lower than that in blank control group (P 0.05). Compared with foam cell group, the expression of m RNA of LXR 偽 and ABCA1 in different concentrations of Ezumab group was increased (P 0.05), and the expression of m RNA was in a concentration dependent manner (P 0.05). Compared with the blank control group, the m RNA and protein expression of LXR 偽 and ABCA1 in foam cell group were decreased (P 0.05). Compared with foam cell group, the m RNA and protein expression of LXR 偽 and ABCA1 in ezumabe 30umol/L group, rosiglitazone group and combined group were increased (P 0.05), and the expression of m RNA and protein in combined group was higher than that in etumabe 30umol/L group and rosiglitazone group, and the expression of m RNA and protein in combined group was higher than that in etumabe group and rosiglitazone group. The expression of m RNA and protein in ABCA1 increased more significantly (P 0.05). Compared with the blank control group, the contents of TC,FC and cholesterol ester (CE) in foam cell group were increased (P 0.05), and the content of CE/TC50%; in foam cell group was higher than that in blank control group. Compared with foam cell group, the content of TC,FC,CE and the ratio of CE/TC in ezumabe 30umol/L group, rosiglitazone group and combination group were decreased (P 0.05), and the ratio of CE/TC in combined group was lower than that in etumabe 30umol/L group and rosiglitazone group. The intracellular TC,FC,CE content and CE/TC ratio decreased more significantly (P 0.05). Conclusion: both ezumab and rosiglitazone can up-regulate LXR 偽-ABCA1 signaling pathway and promote ABCA1-mediated reverse cholesterol transport (RCT), in smooth muscle-derived foam cells to reduce intracellular cholesterol accumulation. The combination of the two can further up-regulate the expression of LXR 偽 and ABCA1, promote cholesterol outflow, and significantly decrease the content of intracellular cholesterol.
【學(xué)位授予單位】:山西醫(yī)科大學(xué)
【學(xué)位級別】:碩士
【學(xué)位授予年份】:2017
【分類號】:R543.5

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相關(guān)期刊論文 前3條

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2 吳佳易;郭志剛;周珊珊;賴文巖;屠燕;;羅格列酮與辛伐他汀聯(lián)用有效抑制動脈粥樣硬化[J];臨床心血管病雜志;2011年01期

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