本文選題：防己黃芪湯　切入點：3T3-L1前脂肪細(xì)胞　出處：《寧夏醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:1.觀察防己黃芪湯對小鼠3T3-L1前脂肪細(xì)胞分化的影響;2.觀察防己黃芪湯對小鼠3T3-L1前脂肪細(xì)胞分化過程中炎癥因子MCP-1、IL-6、TNF-αmRNA表達(dá)的影響。方法:體外培養(yǎng)3T3-L1前脂肪細(xì)胞,在細(xì)胞融合后2天,用傳統(tǒng)“雞尾酒”法(即含胰島素、地塞米松、異丁基-3-甲基黃嘌呤的混合誘導(dǎo)劑)誘導(dǎo)3T3-L1前脂肪細(xì)胞分化為脂肪細(xì)胞。在誘導(dǎo)分化的第0天,設(shè)置溶媒對照組(1 mol/L PBS)和藥物處理組(100μg/ml防己黃芪湯)。1.在成功誘導(dǎo)分化的第10天,用油紅O染色,拍照觀察,用異丙醇溶解油紅O,使用酶標(biāo)儀進(jìn)行定量檢測各組A492nm以間接反映3T3-L1前脂肪細(xì)胞分化程度及細(xì)胞內(nèi)甘油三酯小脂滴含量。2.在成功誘導(dǎo)分化的第10天,提取細(xì)胞總RNA,采用real-time PCR法檢測3T3-L1前脂肪細(xì)胞分化相關(guān)基因,過氧化物酶體增殖受體γ(PPARγ)、脂聯(lián)素(adiponectin)mRNA的表達(dá)情況。3.在成功誘導(dǎo)分化的第10天,提取細(xì)胞總RNA,采用real-time PCR法檢測3T3-L1脂肪細(xì)胞炎癥因子,MCP-1、IL-6、TNF-αmRNA的表達(dá)情況。結(jié)果:1.油紅O染色顯示藥物處理組(100μg/ml防己黃芪湯)中含有甘油三酯小脂滴的成熟脂肪細(xì)胞少于溶媒對照組(1 mol/L PBS),P0.05。2.在誘導(dǎo)分化的第10天,藥物處理組(100μg/ml防己黃芪湯)的PPARγmRNA(P0.005)、脂聯(lián)素mRNA(P0.005)明顯低于溶媒對照組(1 mol/L PBS)。3.在誘導(dǎo)分化的第10天,藥物處理組(100μg/ml防己黃芪湯)的MCP-1 mRNA(P0.05)、TNF-αmRNA(P0.005)明顯低于溶媒對照組(1 mol/L PBS),IL-6 mRNA表達(dá)兩組無統(tǒng)計學(xué)意義。結(jié)論:1.通過定性分析(油紅O染色)、定量分析(3T3-L1前脂肪細(xì)胞分化相關(guān)基因PPARγ、脂聯(lián)素mRNA表達(dá))說明防己黃芪湯可有效抑制3T3-L1前脂肪細(xì)胞分化,減少脂肪細(xì)胞數(shù)量;2.防己黃芪湯可抑制3T3-L1脂肪細(xì)胞中炎癥因子MCP-1、TNF-ɑm RNA的表達(dá),具有改善3T3-L1脂肪細(xì)胞“微炎癥狀態(tài)”,可能對改善脂肪組織胰島素抵抗具有積極作用。
[Abstract]:Objective 1. To observe the effect of Fangji Huangqi decoction on the differentiation of 3T3-L1 preadipocytes in mice 2.To observe the effect of Fangji Huangqi decoction on the expression of MCP-1- IL-6TNF- 偽 mRNA during the differentiation of 3T3-L1 preadipocytes in mice. Methods: 3T3-L1 preadipocytes were cultured in vitro. Two days after cell fusion, 3T3-L1 preadipocytes were induced to differentiate into adipocytes by traditional "cocktail" method (I. e., mixed inducers containing insulin, dexamethasone, isobutyl -3-methylxanthine). On day 0 of differentiation, 3T3-L1 preadipocytes were induced to differentiate into adipocytes. #number0# 渭 g / ml Fangji Astragalus decoction (100 渭 g / ml) was used in the control group and the drug treatment group. On the 10th day after successful differentiation induction, oil red O staining was used to take photos and observe the differentiation. Oil red O4 was dissolved with isopropanol, and A492nm was quantitatively detected by enzymatic analyzer to indirectly reflect the differentiation degree of 3T3-L1 preadipocytes and the content of triglyceride small lipid droplets in the cells. Cell total RNAs were extracted, and the expression of 3T3-L1 preadipocyte differentiation related genes, peroxisome proliferating receptor 緯 -PPAR- 緯, adiponectinine adiponectinine mRNA expression was detected by real-time PCR assay. Total cell RNAs were extracted, and the expression of 3T3-L1 inflammatory cytokine MCP-1and IL-6TNF- 偽 mRNA was detected by real-time PCR method. Results: 1. Oil red O staining showed that the mature adipocytes containing triglyceride small lipid drops in the drug treatment group were less than those in the drug treatment group (100 渭 g / ml Fangji astragalus decoction). In the solvent control group, P0. 05.2 of 1 mol/L PBSs was observed on the 10th day after induction of differentiation. The PPAR 緯 mRNAs (P0.005) and adiponectin mRNAs (P0.005) of 100 渭 g / ml Fangji Astragalus decoction in the drug treatment group were significantly lower than those in the solvent control group (1 渭 g / ml Fangji astragalus decoction). The expression of MCP-1 mRNA- 偽 mRNA-TNF- 偽 mRNA-P0.005 in the drug treatment group was significantly lower than that in the control group (P 0.005). Conclusion: 1: 1. Quantitative analysis of 3T3-L1 preadipocyte differentiation related units by qualitative analysis (oil red O staining). Because of the expression of PPAR 緯, adiponectin mRNA, Fangji Huangqi decoction could effectively inhibit the differentiation of 3T3-L1 preadipocytes. Fangji Huangqi decoction can inhibit the expression of MCP-1and TNF- RNA in 3T3-L1 adipocytes and improve the "microinflammatory state" of 3T3-L1 adipocytes, which may play a positive role in improving insulin resistance in adipose tissue.
【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】：R285.5

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