本文選題：肺血管平滑肌細(xì)胞 + 低氧性肺動脈高壓　； 參考：《寧夏醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:觀察在缺氧條件下進(jìn)行枸杞多糖干預(yù)后肺血管平滑肌細(xì)胞中沉默接合型信息調(diào)節(jié)因子2同源蛋白1(SIRT1)、基質(zhì)金屬蛋白酶9(MMP-9)及缺氧誘導(dǎo)因子1α(HIF-1α)的表達(dá)。方法:肺血管平滑肌細(xì)胞分為常氧培養(yǎng)的正常細(xì)胞(200 m L/L氧氣)、2 mg/m L枸杞多糖處理的常氧細(xì)胞、DMSO處理的常氧細(xì)胞、DMSO處理的缺氧細(xì)胞(100 m L/L氧氣)、(0.5、1、2)mg/m L枸杞多糖處理的缺氧細(xì)胞、SIRT1的特異性激動劑白蘆藜醇(10umol/L)處理的缺氧細(xì)胞、非特異性激動劑SR-1720(1umol/L)處理的缺氧細(xì)胞、SIRT1抑制劑EX-527(0.5umol/L)處理的缺氧細(xì)胞,培養(yǎng)于6、12、24 h,實(shí)時(shí)定量PCR、Western blot法分別檢測SIRT1、MMP-9及HIF-1α的m RNA和蛋白表達(dá)量;在缺氧的肺血管平滑肌細(xì)胞中加入枸杞多糖處理12 h,實(shí)時(shí)定量PCR、Western blot法分別檢測肺血管平滑肌細(xì)胞中SIRT1、MMP-9及HIF-1α的表達(dá);最后按以上分組行劃痕及Transwell小室實(shí)驗(yàn)。結(jié)果:1.缺氧條件下肺血管平滑肌細(xì)胞SIRT1 m RNA和蛋白水平降低,MMP-9及HIF-1αm RNA和蛋白水平升高。2.在缺氧條件下加入LBP可以提高SIRT1的表達(dá),同時(shí)抑制MMP-9及HIF-1α的表達(dá),且此作用具有濃度依賴性,隨著低、中、高LBP濃度梯度的升高SIRT1的表達(dá)量逐漸增多,同時(shí)MMP-9、HIF-1α的表達(dá)量逐漸降低。3.SIRT1的特異性激動劑白藜蘆醇可抑制MMP-9的表達(dá)。4.在缺氧條件下,肺血管平滑肌細(xì)胞的遷移及侵襲能力增強(qiáng),而加入枸杞多糖后抑制了細(xì)胞的侵襲和遷移能力。結(jié)論:1.在缺氧條件下肺血管平滑肌細(xì)胞中SIRT1基因表達(dá)水平下調(diào),而MMP-9及HIF-1α基因表達(dá)水平上調(diào),我們推測在肺血管平滑肌細(xì)胞中SIRT1、MMP-9以及HIF-1α這三者表達(dá)量的變化與低氧性肺動脈高壓的發(fā)生和發(fā)展具有一定的相關(guān)性。2.當(dāng)在缺氧的肺血管平滑肌細(xì)胞中加入低、中、高劑量的LBP后,我們發(fā)現(xiàn)隨著LBP濃度梯度的升高,SIRT1的表達(dá)量上升,而MMP-9及HIF-1α的表達(dá)反而隨著LBP濃度梯度的升高而降低,這些告訴我們LBP可能對于低氧性肺動脈高壓具有一定的改善作用,且它的這種作用主要是通過調(diào)節(jié)SIRT1及相關(guān)因子表達(dá)量的變化來實(shí)現(xiàn)的。3.SIRT1的激動劑白藜蘆醇可以抑制MMP-9的表達(dá),提示我們SIRT1、MMP-9及HIF-1α三者之間的表達(dá)具有一定的相關(guān)性。4.LBP可以抑制缺氧條件下血管平滑肌細(xì)胞的遷移和侵襲,這點(diǎn)告訴我們它可以抑制一些由于缺氧而引起的肺血管增殖以及重塑(如低氧性肺動脈高壓)等疾病的發(fā)生。
[Abstract]:Aim: to observe the expression of silencing signal regulatory factor-2 homologous protein 1 sirt1, matrix metalloproteinase 9 (MMP-9) and hypoxia inducible factor-1 偽 (HIF-1 偽) in pulmonary vascular smooth muscle cells after treatment with Lycium barbarum polysaccharides (LBP) under hypoxia.Methods: pulmonary vascular smooth muscle cells were divided into normal cells cultured in normoxic oxygen (200ml 路L / L) and normal oxygen cells treated with 2 mg/m / L Lycium barbarum polysaccharides (LBP). DMSO-treated normoxic cells treated with DMSO-treated hypoxia cells (100ml / L) were treated with 100ml / L oxygen / L oxygen (0.5mg / mL) / mL Lycium barbarum polysaccharides (LBP).The anoxic cells were treated with 10 umol / L, a specific agonist of SIRT1.Hypoxia cells treated with SR-1720U 1umol/ L) and treated with SIRT1 inhibitor EX-5270.5umol/ L) were cultured for 24 h. The expression of m RNA and protein of SIRT1 MMP-9 and HIF-1 偽 were detected by real-time quantitative PCR Western blot.After treated with Lycium barbarum polysaccharides for 12 h, the expression of SIRT1MMP 9 and HIF-1 偽 in pulmonary vascular smooth muscle cells (VSMCs) were detected by real-time quantitative PCR Western blot assay. Finally, scratches and Transwell chamber experiments were performed according to the above groups.The result is 1: 1.Under hypoxia condition, the levels of SIRT1 m RNA and protein in pulmonary vascular smooth muscle cells decreased, and the levels of MMP-9 and HIF-1 偽 m RNA and protein increased.Under hypoxia condition, LBP could increase the expression of SIRT1 and inhibit the expression of MMP-9 and HIF-1 偽 in a concentration-dependent manner. With the increase of the gradient of LBP concentration, the expression of SIRT1 increased gradually.At the same time, the expression of MMP-9 and HIF-1 偽 decreased gradually. 3. Resveratrol, a specific agonist of SIRT1, inhibited the expression of MMP-9.Under hypoxia condition, the migration and invasion of pulmonary vascular smooth muscle cells were enhanced, but the ability of invasion and migration was inhibited by adding Lycium barbarum polysaccharides.Conclusion 1.The expression of SIRT1 gene was down-regulated in pulmonary vascular smooth muscle cells under hypoxia, while the expression of MMP-9 and HIF-1 偽 genes was up-regulated.We speculate that the changes of the expression of SIRT1 MMP-9 and HIF-1 偽 in pulmonary vascular smooth muscle cells may be related to the occurrence and development of hypoxic pulmonary hypertension.After adding low, medium and high doses of LBP into hypoxic pulmonary vascular smooth muscle cells, we found that the expression of SIRT1 increased with the increase of LBP concentration gradient, while the expression of MMP-9 and HIF-1 偽 decreased with the increase of LBP concentration gradient.These results suggest that LBP may have an improved effect on hypoxic pulmonary hypertension, and this effect is mainly achieved by regulating the expression of SIRT1 and related factors. 3. Resveratrol, an agonist of SIRT1, can inhibit the expression of MMP-9.4. LBP could inhibit the migration and invasion of vascular smooth muscle cells under hypoxia.This suggests that it inhibits pulmonary vascular proliferation and remodeling, such as hypoxic pulmonary hypertension, caused by hypoxia.
【學(xué)位授予單位】：寧夏醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R544.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前8條

1 蔣永亮;聶金;劉蕾;戴愛國;;ING4、PHDs以及HIF-1α在低氧性肺動脈高壓大鼠中的表達(dá)變化[J];中國組織化學(xué)與細(xì)胞化學(xué)雜志;2014年02期

2 盛偉;范文艷;;枸杞多糖對小鼠耐缺氧及抗疲勞能力的影響[J];新鄉(xiāng)醫(yī)學(xué)院學(xué)報(bào);2011年03期

3 孫桂菊;左平國;;枸杞多糖功效研究及應(yīng)用狀況[J];東南大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2010年02期

4 王琦;張曉丹;張靜蕾;;枸杞多糖研究進(jìn)展[J];食品研究與開發(fā);2009年10期

5 鄭云鶴;王小同;徐漫歡;徐浩;王宗敏;龔永生;;HIF和VEGF在低氧性肺動脈高壓中的調(diào)控作用[J];溫州醫(yī)學(xué)院學(xué)報(bào);2007年05期

6 陳麗函;王偉;傅玉才;李吉林;許銘炎;;白藜蘆醇對缺血再灌注心肌細(xì)胞凋亡及沉寂信息調(diào)節(jié)因子2表達(dá)的影響[J];中國臨床康復(fù);2006年19期

7 汪建龍;枸杞多糖藥理作用的研究進(jìn)展[J];時(shí)珍國醫(yī)國藥;2005年10期

8 李曉莉,王斌,劉嘉麟,張聲華;枸杞多糖對小鼠耐常壓缺氧能力的影響[J];營養(yǎng)學(xué)報(bào);2000年04期

，

本文編號：1766957