本文選題：腫瘤壞死因子 + 脂肪細(xì)胞　； 參考：《中南大學(xué)》2007年博士論文

【摘要】： 目的 腫瘤壞死因子α(TNF-α)作為促炎因子對(duì)脂肪細(xì)胞功能有多方面的影響，包括促進(jìn)細(xì)胞分化、脂解及脂肪因子分泌等。本文擬研究TNF-α對(duì)傳代培養(yǎng)的兔皮下脂肪細(xì)胞膽固醇流出功能的影響，及可能的作用機(jī)制。 方法 來(lái)源于兔腹股溝皮下的脂肪細(xì)胞以不同濃度的TNF-α(5，10，20 ng／ml)干預(yù)24小時(shí)。用液體閃爍計(jì)數(shù)儀測(cè)量脂肪細(xì)胞對(duì)氚標(biāo)膽固醇的流出率。用逆轉(zhuǎn)錄多聚酶鏈?zhǔn)椒磻?yīng)(RT-PCR)檢測(cè)過(guò)氧化物酶體增殖物激活受體γ(PPARγ)、肝X受體α(LXRα)及三磷酸腺苷結(jié)合盒轉(zhuǎn)運(yùn)體A1(ABCA1)的mRNA表達(dá)。 結(jié)果 1．5或10 ng／ml TNF-α干預(yù)脂肪細(xì)胞24小時(shí)，可使膽固醇流出率增加，且10 ng／ml TNF-α作用最強(qiáng)(7.03±1.17％)，較對(duì)照組(3.92±0.46％)明顯增高。但20 ng／ml TNF-α反而使膽固醇流出率較10 ng／ml時(shí)下降了16.2％(P＜0.05)。 2．5或10 ng／ml TNF-α可上調(diào)ABCA1表達(dá)，其表達(dá)量分別為0.44±0.17和0.91±0.26，與對(duì)照組0.20±0.06相比差異有顯著性。20 ng／ml TNF-α使ABCA1表達(dá)有所下降(0.69±0.13)。與此同時(shí)，TNF-α可調(diào)節(jié)脂肪細(xì)胞PPARγ和LXRα表達(dá)。在TNF-α≤10 ng／ml時(shí)起促進(jìn)作用，在20 ng／ml時(shí)起抑制作用。 3．預(yù)先以PPARγ特異性抑制劑GW9662孵育半小時(shí)，可抑制10 ng／ml TNF-α對(duì)PPARγ(0.88±0.22 vs 1.37±0.25，P＜0.05)和LXRα(0.51±0.14 vs 0.32±0.08，P＜0.05)的誘導(dǎo)作用。 結(jié)論 TNF-α影響脂肪細(xì)胞膽固醇流出及其ABCA1表達(dá)，這種作用與TNF-α對(duì)脂肪細(xì)胞PPARγ-LXRα的雙重調(diào)節(jié)作用有關(guān)。 目的 脂肪細(xì)胞肥大伴隨一系列功能改變。我們擬研究在氧化低密度脂蛋白(oxLDL)誘導(dǎo)脂肪細(xì)胞負(fù)荷脂質(zhì)后，TNF-α對(duì)其膽固醇流出和ABCA1表達(dá)的影響是否會(huì)發(fā)生改變。 方法 用50μg／ml oxLDL孵育脂肪細(xì)胞24小時(shí)制造荷脂模型。再以不同濃度TNF-α(5，，10，20 ng／ml)干預(yù)24小時(shí)，觀察膽固醇流出變化及用RT-PCR法檢測(cè)脂肪細(xì)胞PPARγ、LXRα和ABCA1 mRNA表達(dá)。 結(jié)果 1．OxLDL可使脂肪細(xì)胞膽固醇流出增加(7.46±2.34％vs 3.92±0.46％)，PPARγ(1.07±0.26 vs 0.48±0.12)、LXRα(0.83±0.16vs 0.32±0.08)和ABCA1(1.19±0.32 vs 0.20±0.06)表達(dá)上升，P均＜0.05。 2．TNF-α抑制荷脂脂肪細(xì)胞膽固醇流出，10 ng／ml時(shí)作用最強(qiáng)(6.02±0.31％vs3.92±0.46％)，在20 ng／ml時(shí)脂肪細(xì)胞膽固醇流出率略有回升(6.12±0.53)，差異有顯著性。 3．從5 ng／ml到20 ng／ml TNF-α，PPARγ表達(dá)量分別為0.93±0.21，0.77±0.15，0.60±0.16與oxLDL對(duì)照組1.07±0.26相比表達(dá)降低。LXRα表達(dá)量為0.72±0.19，0.61±0.17，0.49±0.09與oxLDL對(duì)照組0.83±0.16相比也有降低。TNF-α呈濃度依賴性抑制脂肪細(xì)胞PPARγ和LXRα表達(dá)。5 ng／ml和10 ng／ml TNF-α分別使脂肪細(xì)胞ABCA1表達(dá)下降了14.4％和34.5％，20ng／ml TNF-α使ABCA1表達(dá)有所回升。 結(jié)論 在oxLDL誘導(dǎo)荷脂的脂肪細(xì)胞中，TNF-α對(duì)PPARγ和LXRα表達(dá)主要表現(xiàn)為抑制作用，并在一定程度上影響到ABCA1表達(dá)及脂肪細(xì)胞膽固醇流出。 目的 姜黃素是一類中藥提取物，已證實(shí)其具有抗炎、抗氧化及改善血脂異常等作用。本研究觀察了姜黃素對(duì)脂肪細(xì)胞抗炎脂肪因子脂聯(lián)素表達(dá)和分泌的影響，初步評(píng)價(jià)姜黃素對(duì)脂肪細(xì)胞功能的影響。 方法 新西蘭大白兔腹股溝皮下脂肪細(xì)胞以不同濃度姜黃素(5，10，20μg／ml)孵育24小時(shí)，用ELISA試劑盒檢測(cè)細(xì)胞培養(yǎng)上清液脂聯(lián)素水平，并用RT-PCR方法檢測(cè)脂聯(lián)素及其上游因子PPARγ的表達(dá)。 結(jié)果 5μg／ml至20μg／ml姜黃素分別使兔皮下脂肪細(xì)胞脂聯(lián)素分泌水平較對(duì)照組(3.13±0.21μmol／l)增加(3.93±0.26，4.49±0.34，5.21±0.39μmol／l)。脂聯(lián)素mRNA表達(dá)也較對(duì)照組(0.40±0.10)增加(0.55±0.09，0.66±0.11，0.80±0.13)。姜黃素促進(jìn)PPARγ表達(dá)的作用與脂聯(lián)素表達(dá)的改變一致，并在20μg／ml時(shí)達(dá)到最大(1.01±0.15vs 0.48±0.12，P＜0.05)。 結(jié)論 姜黃素可促進(jìn)兔皮下脂肪細(xì)胞脂聯(lián)素的表達(dá)和分泌，這種作用與其上調(diào)脂肪細(xì)胞PPARγ表達(dá)有關(guān)。
[Abstract]:objective
Tumor necrosis factor alpha (TNF- alpha), as a pro-inflammatory factor, has many effects on the function of adipocyte, including promoting cell differentiation, lipo secretion and adipose factor secretion. This paper intends to study the effect of TNF- alpha on the cholesterol efflux function of subcutaneous adipocytes in subcutaneous subcutaneous tissue and the possible mechanism of action.
Method
The adipocytes derived from the rabbit groin were interfered with different concentrations of TNF- alpha (5,10,20 ng / ml) for 24 hours. The efflux rate of tritium cholesterol was measured by the liquid scintillation counting instrument. The peroxisome proliferator activated receptor gamma (PPAR gamma), the liver X receptor alpha (LXR a) and the three phosphoric acid were detected by the reverse transcription polymerase chain reaction (RT-PCR). MRNA expression of adenosine binding cassette transporter A1 (ABCA1).
Result
1.5 or 10 ng / ml TNF- alpha could increase the cholesterol efflux rate for 24 hours, and 10 ng / ml TNF- alpha was strongest (7.03 + 1.17%), which was significantly higher than that in the control group (3.92 + 0.46%), but the cholesterol efflux rate decreased by 16.2% (P < 0.05) compared to 10 ng / ml.
2.5 or 10 ng / ml TNF- alpha could up regulate the expression of ABCA1, and the expression amount was 0.44 + 0.17 and 0.91 + 0.26 respectively. Compared with the control group 0.20 + 0.06, there was a significant difference between.20 ng / ml TNF- a to decrease the ABCA1 expression (0.69 + 0.13). Meanwhile, TNF- a could regulate the expression of PPAR gamma and LXR alpha in adipocytes. The inhibitory effect of 20 ng / ml.
3. the induction effect of 10 ng / ml TNF- alpha on PPAR gamma (0.88 + 0.22 vs 1.37 + 0.25, P < 0.05) and LXR alpha (0.51 + 0.14 vs 0.32 + 0.08, P < 0.05) can be inhibited by pre incubation of PPAR gamma specific inhibitor.
conclusion
TNF- alpha affects the cholesterol efflux and ABCA1 expression in adipocytes, which is related to the dual regulation of TNF- alpha on PPAR -LXR -LXR of adipocytes.
objective
Adipocyte hypertrophy is accompanied by a series of functional changes. We intend to study whether the effect of TNF- alpha on the cholesterol efflux and the expression of ABCA1 after the oxidative low density lipoprotein (oxLDL) induced lipid cell load may change.
Method
Fat cells were incubated with 50 g / ml oxLDL for 24 hours to produce a fat charge model, and then 24 hours were intervened with different concentrations of TNF- alpha (5,10,20 ng / ml). The changes in cholesterol efflux were observed and the expression of PPAR gamma, LXR A and ABCA1 mRNA were detected by RT-PCR method.
Result
1.OxLDL could increase the cholesterol efflux of adipocytes (7.46 + 2.34%vs 3.92 + 0.46%), PPAR gamma (1.07 + 0.26 vs 0.48 + 0.12), LXR alpha (0.83 + 0.16vs 0.32 + 0.08) and ABCA1 (1.19 + 0.32 vs 0.20 +%) and P < 0.05.
2.TNF- alpha inhibited the cholesterol efflux of fat charged fat cells. The effect of 10 ng / ml was the strongest (6.02 + 0.31%vs3.92 + 0.46%). The cholesterol efflux rate of adipocytes was slightly increased at 20 ng / ml (6.12 + 0.53), and the difference was significant.
3. from 5 ng / ml to 20 ng / ml TNF- a, PPAR gamma expression was 0.93 + 0.21,0.77 + 0.16 compared with oxLDL control group 1.07 + 0.26, and the expression of.LXR a decreased to 0.72 + 0.19,0.61 + 0.17,0.49 + 0.09 and 0.83 + 5. Ng / ml and 10 ng / ml TNF- alpha reduced the expression of ABCA1 in adipocytes by 14.4% and 34.5% respectively, and 20ng / ml TNF- increased the expression of ABCA1.
conclusion
In the adipocytes induced by oxLDL, the expression of TNF- alpha on PPAR gamma and LXR alpha is mainly inhibited, and to a certain extent, it affects the expression of ABCA1 and the cholesterol efflux of adipocytes.
objective
Curcumin is a kind of traditional Chinese medicine extract, which has been proved to have anti-inflammatory, antioxidation and improvement of blood lipid abnormality. This study observed the effect of curcumin on the expression and secretion of adipocyte adiponectin in adipocytes, and preliminarily evaluated the effect of curcumin on the function of adipocytes.
Method
The subcutaneous fat cells in the groin of New Zealand rabbits were incubated with curcumin (5,10,20 / g / ml) for 24 hours. The level of adiponectin in cultured supernatant was detected by ELISA kit, and the expression of adiponectin and its upstream factor PPAR y was detected by RT-PCR.
Result
5 g / ml to 20 g / ml curcumin increased adiponectin secreting level in subcutaneous adipocytes (3.13 + 0.21 Mu mol / L) (3.93 + 0.26,4.49 + 0.34,5.21 + 0.39 UU / L). The mRNA expression of adiponectin was also increased (0.40 + 0.10) (0.55 + 0.09,0.66 + 0.13). The change of expression was consistent, and reached the maximum at 20 g / ml (1.01 + 0.15vs 0.48 + 0.12, P < 0.05).
conclusion
Curcumin can promote the expression and secretion of adiponectin in rabbit subcutaneous adipocytes, which is related to upregulated the expression of PPAR gamma in adipocytes.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2007
【分類號(hào)】：R589.2;R363

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