本文選題：游離脂肪酸 + 高分子量透明質(zhì)酸��； 參考：《寧夏醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的探討透明質(zhì)酸(Hyaluronic acid,HA)對(duì)游離脂肪酸(Free fatty acid,FFA)引起的人主動(dòng)脈血管平滑肌細(xì)胞(Human aortic vascular smooth muscle cells,HA-VSMC)表達(dá)白介素-6(Interleukin-6,IL-6)、白細(xì)胞介素-8(Interleukin-8,IL-8)和腫瘤壞死因子-α(Tumor necrosis factorα,TNF-α)的影響,為對(duì)血管病變的體內(nèi)研究和臨床治療提供理論和實(shí)驗(yàn)依據(jù)。方法常規(guī)培養(yǎng)人HA-VSMC細(xì)胞系,采用不同濃度的FFA(0、100、200、400μmol/L)、不同濃度FFA(0、100、200、400μmol/L)+高分子量透明質(zhì)酸(High molecular weight hyaluronic acid,HMW-HA,1mg/ml)和不同濃度FFA(0、100、200、400μmol/L)+低分子量透明質(zhì)酸(Low molecular weight hyaluronic acid,LMW-HA,1mg/ml)處理細(xì)胞24、48h;采用MTT法檢測(cè)FFA、FFA+LMW-HA和FFA+HMW-HA對(duì)HA-VSMC細(xì)胞存活率的影響;采用Ed U染色檢測(cè)不同處理后細(xì)胞增殖活性;采用ELISA試劑盒檢測(cè)培養(yǎng)上清IL-6、IL-8和TNF-α含量;采用實(shí)時(shí)熒光定量PCR法檢測(cè)IL-6、IL-8和TNF-αm RNA表達(dá)水平;蛋白印記法(Western blot)檢測(cè)IL-6、IL-8和TNF-α蛋白水平的表達(dá)。結(jié)果1、FFA在400μmol/L內(nèi)可誘導(dǎo)HA-VSMC增殖且有時(shí)間和濃度依賴性;2、HMW-HA、LMW-HA均對(duì)FFA誘導(dǎo)的HA-VSMC增殖有抑制作用,且HMW-HA抑制作用強(qiáng)于LMW-HA;3、FFA單獨(dú)和聯(lián)合高低分子量HA處理細(xì)胞6、12、24、48h后細(xì)胞培養(yǎng)上清IL-6、IL-8和TNF-α隨培養(yǎng)濃度和時(shí)間的增加而增加,HA可以抑制上述作用;4、熒光定量PCR和Western blot結(jié)果顯示FFA+HMW-HA組和FFA+LMW-HA組IL-6、IL-8和TNF-αm RNA及其蛋白表達(dá)水平均低于FFA組,其中FFA+HMW-HA組又低于FFA+LMW-HA組,差異均有統(tǒng)計(jì)學(xué)意義(均P0.01)。結(jié)論(1)游離脂肪酸在400μmol/L內(nèi)可誘導(dǎo)HA-VSMC增殖且有濃度依賴性;(2)HA能抑制FFA引起的HA-VSMC增殖,且HMW-HA抑制作用強(qiáng)于LMW-HA;(3)FFA顯著促進(jìn)HA-VSMC IL-6、IL-8和TNF-α的分泌,且有明顯的時(shí)間和濃度依耐性;(4)FFA誘導(dǎo)HA-VSMC細(xì)胞IL-6、IL-8和TNF-αm RNA和蛋白表達(dá)升高;(5)HA對(duì)FFA引起的HA-VSMC細(xì)胞表達(dá)IL-6、IL-8和TNF-α有抑制作用,下調(diào)IL-6、IL-8和TNF-αm RNA和蛋白表達(dá),且HMW-HA抑制效果強(qiáng)于LMW-HA。
[Abstract]:Objective to investigate the expression of Hyaluronic acid (HA) in human aortic vascular smooth muscle cells induced by Free fatty acid (FFA) and to express interleukin and tumor necrosis factor alpha. The effect of TNF- alpha, providing theoretical and experimental basis for the study and clinical treatment of vascular lesions. Methods routine culture of human HA-VSMC cell lines, using different concentrations of FFA (0100200400 mu mol/L), different concentrations of FFA (0100200400 mol/L) + high molecular weight hyaluronic acid (High molecular weight hyaluronic acid, HMW-HA, etc.) Different concentrations of FFA (0100200400 mu mol/L) + low molecular weight hyaluronic acid (Low molecular weight hyaluronic acid, LMW-HA, 1mg/ml) were used to treat cell 24,48h, and MTT method was used to detect the survival of the cells. The content of IL-6, IL-8 and TNF- alpha was raised and the RNA expression level of IL-6, IL-8 and TNF- alpha m was detected by real time fluorescence quantitative PCR method, and protein imprinting method (Western blot) was used to detect IL-6, expression and expression of alpha protein. Results 1 The proliferation of MC has a inhibitory effect, and the inhibitory effect of HMW-HA is stronger than that of LMW-HA; 3, FFA alone and combined high and low molecular weight HA cells 6,12,24,48h cell culture supernatant IL-6, IL-8 and TNF- alpha increase with the increase of culture concentration and time, HA can inhibit the effect. 4, fluorescent quantitative PCR and Western results show the group and the Western The expressions of IL-6, IL-8 and TNF- alpha m RNA and their protein expressions were lower than those in the FFA group, and the FFA+HMW-HA group was lower than the FFA+LMW-HA group. The difference was statistically significant (all P0.01). Conclusion (1) the free fatty acids can induce HA-VSMC proliferation and concentration dependence in 400 mu mol/L. (2) HA can inhibit the proliferation and the inhibitory effect is stronger than that of the FFA+LMW-HA. (3) (3) FFA significantly promoted the secretion of HA-VSMC IL-6, IL-8 and TNF- alpha, and had obvious time and concentration dependent tolerance; (4) FFA induced IL-6 in HA-VSMC cells, IL-8 and TNF- alpha m and protein expression increased; (5) The effect of the system is better than LMW-HA.
【學(xué)位授予單位】：寧夏醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R587.2;R54

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