發(fā)布時間：2018-04-15 08:31

  本文選題：血管鈣化 + 姜黃素��； 參考：《中山大學(xué)》2016年碩士論文

【摘要】：研究背景:血管鈣化(vascular calcification)指的是鈣鹽沉積于動脈管壁的過程,它可以導(dǎo)致動脈管壁變硬,順應(yīng)性降低,在臨床上常見于動脈粥樣硬化,慢性腎病和糖尿病患者,嚴(yán)重危害人類的健康。然而,由于對血管鈣化的發(fā)生和發(fā)展機(jī)制缺乏深入的認(rèn)識,目前臨床上缺乏治療血管鈣化的有效方法。因此,研究血管鈣化的病因和發(fā)生發(fā)展的病理機(jī)制,闡明調(diào)節(jié)血管鈣化的分子機(jī)制非常重要。近些年來的大量研究發(fā)現(xiàn):血管鈣化的過程類似于骨的生成,同樣,也是一種受基因表達(dá)調(diào)控的,主動的過程。血管鈣化的主要細(xì)胞來源是血管平滑肌細(xì)胞(vascular smooth muscle cells,VSMCs)。在損傷因素如鈣磷代謝紊亂、氧化應(yīng)激的刺激下,血管平滑肌細(xì)胞可從正常的收縮表型轉(zhuǎn)換為成骨樣表型并分泌大量的骨相關(guān)蛋白:堿性磷酸酶(alkaline phosphatase,ALP)、同源盒基因(Msx2)、核心結(jié)合因子α1(cbfa1/Runx2)、骨形態(tài)發(fā)生蛋白(bone morphogenetic proteins,BMPs)、成骨相關(guān)轉(zhuǎn)錄因子(Osterix)、骨鈣素(OCN)等,從而促進(jìn)細(xì)胞鈣化。因此血管平滑肌細(xì)胞的成骨樣分化過程與血管鈣化的有著密不可分的關(guān)系。大量研究已證實了鈣磷代謝紊亂是導(dǎo)致血管平滑肌細(xì)胞鈣化和凋亡的主要因素之一。血管平滑肌細(xì)胞凋亡在血管鈣化的過程中發(fā)揮了重要作用。在血管平滑肌鈣化之前可檢測到凋亡小體的出現(xiàn),血管平滑肌細(xì)胞釋放凋亡小體可啟動血管鈣化,細(xì)胞鈣化受血管平滑肌細(xì)胞凋亡調(diào)節(jié),通過抑制血管平滑肌細(xì)胞的凋亡可抑制其鈣化的程度。我們以前的研究也證實:平滑肌細(xì)胞凋亡可調(diào)節(jié)氧化型低密度脂蛋白誘導(dǎo)的細(xì)胞鈣化。姜黃素(Curcumin),是一種來自植物姜黃的提取物,可減輕血管病變包括血管損傷后的內(nèi)膜新生和動脈粥樣硬化。姜黃素對血管平滑肌細(xì)胞具有不同的藥理作用:抗炎,抗氧化,抑制細(xì)胞增殖和遷移等。此外,姜黃素可抑制細(xì)胞凋亡和成骨細(xì)胞的鈣化。然而,迄今為止,姜黃素對血管平滑肌細(xì)胞的成骨樣分化和鈣化的影響未見報道。我們推測姜黃素可能通過抑制血管平滑肌細(xì)胞凋亡來影響血管平滑肌細(xì)胞成骨樣分化及鈣化。本研究采用體外血管鈣化模型,采用高鈣高磷處理體外培養(yǎng)的血管平滑肌細(xì)胞,誘導(dǎo)細(xì)胞鈣化,并用姜黃素處理細(xì)胞,觀察姜黃素對血管平滑肌細(xì)胞成骨樣分化和鈣化的影響及其凋亡信號機(jī)制。目的:1.姜黃素是否抑制血管平滑肌細(xì)胞成骨樣分化和鈣化。2.姜黃素是否抑制血管平滑肌細(xì)胞凋亡。3.姜黃素抑制血管平滑肌細(xì)胞鈣化的凋亡信號通路是否與JNK/Bax信號通路有關(guān)。方法:本研究采用10m M BGP和3m M氯化鈣處理血管平滑肌細(xì)胞3天和10天后,用茜素紅染色測細(xì)胞鈣化,鄰甲酚酞絡(luò)合劑法測鈣離子濃度,檢測ALP活性。用q PCR檢測血管平滑肌細(xì)胞成骨樣分化的相關(guān)分子和凋亡基因的表達(dá)水平。用Western blot檢測JNK/Bax的表達(dá)水平。用流式細(xì)胞技術(shù)和Caspase3活性檢測的方法檢測了血管平滑肌細(xì)胞凋亡。結(jié)果:姜黃素可抑制高鈣高磷誘導(dǎo)的血管平滑肌細(xì)胞成骨樣分化和鈣化,同時可抑制血管平滑肌細(xì)胞凋亡。另外,姜黃素可下調(diào)p-JNK、Bax的表達(dá)水平。抑制JNK信號能明顯阻斷血管平滑肌細(xì)胞成骨樣分化和鈣化。結(jié)論:姜黃素抑制了血管平滑肌細(xì)胞成骨樣分化和鈣化,其機(jī)制很可能與JNK/Bax凋亡信號有關(guān)。
[Abstract]:Background: vascular calcification (vascular calcification) refers to the process of calcium deposition in the arterial wall, it can lead to arterial wall hardening, reduced compliance in clinical common in atherosclerosis, chronic kidney disease and diabetes, serious harm to human health. However, because of the occurrence and development mechanism of vascular calcification the lack of deep understanding of the current clinical lack of effective method for the treatment of vascular calcification. Therefore, the etiology of vascular calcification and the occurrence and development of pathological mechanism, to elucidate the molecular mechanisms regulating vascular calcification is very important. A large number of studies in recent years found that the generation process of vascular calcification is similar to bone of the same, is also a the regulation of gene expression, active process. The main source of cells for vascular calcification in vascular smooth muscle cells (vascular smooth muscle cells, VSMCs). The damage factors such as calcium and phosphorus generation Xie disorder, oxidative stress stimulation, vascular smooth muscle cells from normal contractile phenotype into osteoblast like phenotype and secretion of bone related protein A: alkaline phosphatase (alkaline phosphatase, ALP), homeobox gene (Msx2), core binding factor alpha 1 (cbfa1/ Runx2), bone morphogenetic protein (bone morphogenetic proteins, BMPs), bone related transcription factor (Osterix), osteocalcin (OCN), so as to promote cell calcification. Therefore bone differentiation and vascular calcification into vascular smooth muscle cells has a close relationship. A large number of studies have proved that the metabolism of calcium and phosphorus is one of the main causes of vascular calcification and apoptosis smooth muscle cells. The apoptosis of vascular smooth muscle cells play an important role in the process of vascular calcification in vascular smooth muscle. Before calcification can be detected by the appearance of apoptotic bodies, vascular smooth muscle cells to release. Dead bodies can be initiated by cell vascular calcification, calcification of vascular smooth muscle cell apoptosis regulation by inhibiting the apoptosis of vascular smooth muscle cells can inhibit the calcification. Our previous studies have confirmed that apoptosis of smooth muscle cells can regulate the oxidized low density lipoprotein induced cell calcification. Curcumin (Curcumin), is a kind of plant from turmeric the extract can reduce the vascular lesions including neointima after vascular injury and atherosclerosis. Curcumin has different pharmacological effects on vascular smooth muscle cells: anti-inflammatory, antioxidant, inhibit the proliferation and migration of cells. In addition, curcumin can inhibit the apoptosis of osteoblasts and calcification. However, so far, the effect of curcumin on vascular smooth muscle cells the differentiation and calcification of bone like effects have not been reported. We speculate that curcumin may influence by inhibiting vascular smooth muscle cell apoptosis Vascular smooth muscle cell differentiation and calcification of bone samples. This study used vascular calcification of vascular smooth muscle cells by in vitro model, high calcium and high phosphorus treatment in vitro, induce cell calcification, and cells treated with curcumin, the effects of curcumin on vascular smooth muscle cells of osteogenic differentiation and calcification effect and apoptosis signaling mechanism. Objective: 1. whether curcumin inhibits vascular smooth muscle cells is the apoptosis signaling pathway of bone like differentiation and calcification of.2. whether curcumin inhibited the apoptosis of vascular smooth muscle cells.3. curcumin inhibits calcification in vascular smooth muscle cells associated with the JNK/ Bax pathway. Methods: This study used 10m M BGP and 3M M calcium chloride treatment of vascular smooth muscle cells in 3 days and 10 days were measured calcification by alizarin red staining, measured the concentration of calcium ion o-cresolphthalein complexone method, ALP activity detection. PCR detection of vascular smooth muscle cells Q osteoblast like points The expression level of apoptosis related molecules and genes. The expression level of Western blot in the detection of JNK/Bax. The apoptosis of vascular smooth muscle cells were detected by flow cytometry and Caspase3 assay. Results: Curcumin could inhibit vascular smooth muscle cells induced by high calcium and high phosphorus into differentiation and calcification of bone, and can inhibit the apoptosis of vascular the smooth muscle cells. In addition, curcumin suppressed the p-JNK expression level of Bax. The inhibition of JNK signaling can markedly inhibit vascular smooth muscle cell differentiation and calcification of bone samples. Conclusion: Curcumin inhibits vascular smooth muscle cell differentiation and calcification of bone, its mechanism may be related to the apoptosis of JNK/Bax signal.

【學(xué)位授予單位】：中山大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R543
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本文編號：1753357