本文關(guān)鍵詞： 高糖 人臍靜脈內(nèi)皮細(xì)胞 球型脂聯(lián)素 一氧化氮 不對稱二甲基精氨酸　出處：《南華大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

【摘要】：目的： 探討球型脂聯(lián)素（gad）對高糖環(huán)境下人臍靜脈內(nèi)皮細(xì)胞（HUVECs）NO及DDAH-ADMA系統(tǒng)的影響 方法： 體外培養(yǎng)HUVECs至對數(shù)生長期，gad(2.5ug/ml)處理高糖誘導(dǎo)不同時(shí)間（24h、48h）HUVECs，，不同濃度gad（0.1、0.5、2.5、12.5μg/ml）處理高糖誘導(dǎo)48h HUVECs，硝酸還原酶法測定培養(yǎng)液上清NO濃度；再將HUVECs隨機(jī)分為正常對照組（5.5mmol/L葡萄糖）、高糖組（33.0mmol/L）、gad組（33.0mmol/L葡萄糖+2.5ug/ml gad），L-NAME組（33.0mmol/L葡萄糖+2.5μg/ml gad+100μmol/LL-NAME），硝酸還原酶法檢測培養(yǎng)液上清NO濃度，化學(xué)比色法測定NOS活力，高效液相色譜法分析ADMA水平，根據(jù)ADMA的量反映DDAH活性，westernblot檢測eNOS、P-eNOS的表達(dá)。 結(jié)果： ①gad處理高糖誘導(dǎo)HUVECs不同時(shí)間（24h、48h），高糖作用24h，引起NO分泌增加，而作用48h，NO分泌顯著減少（P0.05），與同一時(shí)間高糖組相比，gad組NO顯著升高（P0.05）；②不同濃度gad（0.1、0.5、2.5、12.5μg/ml）處理高糖誘導(dǎo)48h HUVECs，NO水平先隨gad濃度的增加而顯著增加，但gad2.5μg/mL與12.5μg/mL相比，NO水平差異無顯著性（P0.05）；③2.5μg/ml gad處理高糖誘導(dǎo)48h HUVECs，高糖組較對照組相比，NO含量顯著降低，NOS活性及p-eNOS表達(dá)均減少（P0.05），而總eNOS表達(dá)無顯著差異（P0.05）；④高糖組與正常對照組相比ADMA含量升高，DDAH活性降低，差異均有統(tǒng)計(jì)學(xué)意義（P0.05），2.5μg/mL gad處理后，與高糖組相比，NO含量顯著升高，NOS活性顯著增強(qiáng)，P-eNOS表達(dá)顯著增加，總eNOS蛋白表達(dá)仍無統(tǒng)計(jì)學(xué)差異（P0.05）；ADMA水平gad組與高糖組相比降低，DDAH活性增加，差異均有統(tǒng)計(jì)學(xué)意義（P0.05），加入L-NAME100μmol/L，與gad組相比，NO含量顯著減少，NOS活性減低，P-eNOS表達(dá)減少，差異有統(tǒng)計(jì)學(xué)差異（P0.05），總eNOS表達(dá)仍無差異（P0.05），ADMA水平升高，DDAH活性降低，差異均具有顯著性（P0.05）。 結(jié)論： 1、高糖環(huán)境下gad可逆轉(zhuǎn)HUVECs合成NO的減少，并升高NOS活性，增加eNOS-Ser1177磷酸化，但對eNOS總蛋白表達(dá)并無影響。 2、高糖環(huán)境下gad可升高DDAH活性，降低ADMA水平，而這種作用可能依賴于gad升高NO水平，恢復(fù)NOS活性；同時(shí)，DDAM活性升高以及ADMA水平下降又可升高NO水平，增加NOS活性。
[Abstract]:Objective:. To investigate the effects of globular adiponectin on the nitric oxide (no) and DDAH-ADMA system of human umbilical vein endothelial cells (HUVECs) in high glucose environment. Methods:. HUVECs were incubated in vitro with HUVECs to logarithmic growth stage (2.5ugml) for 24 h / 48h after high glucose induction, and 0.5 渭 g / ml (0.5 渭 g / ml) for 48h after high glucose induction. Nitric acid reductase method was used to determine the concentration of nitric oxide (no) in the supernatant of cultured HUVECs. HUVECs was randomly divided into normal control group (5.5mmol / L glucose), high glucose group (33.0mmol / L) and high glucose group (33.0 mmol / L glucose 2.5ug-ml) -NAME group. Nitric acid reductase method was used to detect no concentration in the culture medium, and the activity of NOS was determined by chemical colorimetry. High performance liquid chromatography (HPLC) was used to analyze the level of ADMA and to detect the expression of eNOSn P-Enos by Western blot according to the activity of DDAH reflected by the quantity of ADMA. Results:. 1 g ad treatment with high glucose induced HUVECs at different time points for 24 h or 48 h, and high glucose for 24 h resulted in the increase of no secretion. Compared with the high glucose group at the same time, the no level in the Gad group was significantly higher than that in the high glucose group at the same time. The level of no increased significantly with the increase of the concentration of gad at 48h after treatment with 0.5 渭 g / ml of 0.5 渭 g / ml of gad. However, there was no significant difference in the level of no between gad2.5 渭 g / mL and 12.5 渭 g / mL in the treatment of HUVECs induced by high glucose for 48 h with 32.5 渭 g / ml gad. Compared with the control group, the content of nitric oxide monoxide synthase (no) and the expression of p-Enos in the high glucose group were significantly lower than those in the control group. There was no significant difference in the expression of total eNOS between the hyperglycemic group and the normal group. Compared with the control group, the ADMA content increased and the activity of DDAH decreased. After treatment with 2.5 渭 g / mL gad, the content of nitric oxide (no) increased significantly and the activity of nitric oxide synthase increased significantly (P 0.05). The expression of total eNOS protein was not significantly different between gad group and high glucose group, and the activity of gad group was significantly lower than that of high glucose group. Compared with gad group, the content of no decreased significantly and the expression of P-Enos decreased significantly (P 0.05). There was no significant difference in the expression of total eNOS. The level of total eNOS increased and the activity of DDAH decreased, and the difference was significant (P 0.05). Conclusion:. 1. Gad could reverse the decrease of no synthesis in HUVECs, increase the activity of NOS and increase the phosphorylation of eNOS-Ser1177, but had no effect on the expression of total eNOS protein. (2) gad could increase the activity of DDAH and decrease the level of ADMA in high glucose environment, which might depend on gad to increase the level of no and restore the activity of NOS, while the increase of activity of DDAM and the decrease of ADMA could increase the level of no and increase the activity of NOS.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R363

【共引文獻(xiàn)】

相關(guān)期刊論文 前5條

1 周強(qiáng);吳曉蔓;;脂聯(lián)素與肥胖癥、胰島素抵抗及2型糖尿病[J];國際檢驗(yàn)醫(yī)學(xué)雜志;2008年10期

2 許軼洲;高炎;;脂聯(lián)素與心血管疾病[J];生命科學(xué);2010年04期

3 劉蕾;李強(qiáng)翔;;脂聯(lián)素對心血管病變影響的研究進(jìn)展[J];中華高血壓雜志;2010年09期

4 鐘立;羅梅;夏偉;楊淑敏;李啟富;;色素上皮衍生因子對人HepG2肝細(xì)胞脂代謝的影響[J];重慶醫(yī)科大學(xué)學(xué)報(bào);2012年06期

5 袁愛紅;蔡輝;;針刺對高脂大鼠主動(dòng)脈脂聯(lián)素受體基因表達(dá)的影響[J];中西醫(yī)結(jié)合心腦血管病雜志;2010年10期

相關(guān)博士學(xué)位論文 前3條

1 鐘立;色素上皮衍生因子與脂代謝及胰島素抵抗相關(guān)性研究[D];重慶醫(yī)科大學(xué);2011年

2 肖揚(yáng);脂肪酸結(jié)合蛋白與2型糖尿病患者亞臨床動(dòng)脈粥樣硬化的關(guān)系[D];中南大學(xué);2010年

3 劉少偉;非致命性創(chuàng)傷后心肌缺血/再灌注損傷加重的分子機(jī)制及脂聯(lián)素的作用研究[D];第四軍醫(yī)大學(xué);2010年

相關(guān)碩士學(xué)位論文 前1條

1 何姜;重組脂聯(lián)素對氧化應(yīng)激狀態(tài)下內(nèi)皮細(xì)胞NF-κB、iNOS和NO信號(hào)表達(dá)的影響[D];福建醫(yī)科大學(xué);2009年

本文編號(hào)：1496797