發(fā)布時(shí)間：2018-07-11 13:47

  本文選題：肥胖 + 丹酚酸B　； 參考：《北京中醫(yī)藥大學(xué)》2017年博士論文

【摘要】：一、降糖消渴顆�；钛鼋M分對(duì)高脂飲食誘導(dǎo)肥胖C57BL/6J小鼠糖脂代謝的影響目的本研究采用高脂飼料誘導(dǎo)的C57BL/6J肥胖小鼠模型,觀察降糖消渴顆粒活血化瘀組分丹酚酸B、姜黃素對(duì)肥胖及肥胖相關(guān)的糖脂代謝異常的影響;并通過對(duì)肥胖C57BL/6J小鼠脂肪組織中脂肪分解及成脂分化關(guān)鍵轉(zhuǎn)錄因子指標(biāo)的檢測(cè),進(jìn)一步探討降糖消渴顆粒活血化瘀組分在調(diào)控脂肪組織功能,改善機(jī)體糖脂代謝方面的可能作用機(jī)制。方法6周齡C57BL/6J雄性小鼠,共70只,按體重隨機(jī)分為正常對(duì)照組(n=10)及高脂組(n=60),正常對(duì)照組小鼠喂飼普通全價(jià)飼料,高脂組(HFD)小鼠喂飼純化高脂飼料。12周后,以體重超過正常飼料組平均體重的20%為肥胖模型.成功標(biāo)準(zhǔn),將造模成功的肥胖小鼠按體重隨機(jī)分為模型組、二甲雙胍組、丹酚酸B姐、姜黃素組(n=1 1)。各組按10g.BW/O.1ml體積灌胃給藥,丹酚酸B劑量為:100mg/kg.BW/day;姜黃素劑量為:50mg/kg.BW/day;二甲雙胍給藥量為75mg/kg.BW/day,模型組及正常組灌胃等量去離子水,共給藥8周。每周同一時(shí)間監(jiān)測(cè)小鼠體重、攝食量、空腹血糖(FBG),在第4、8周測(cè)定各組小鼠體脂含量及口服葡萄糖耐量(OGTT)。實(shí)驗(yàn)結(jié)束后取材,檢測(cè)血中甘汕三酯(TG)、總膽固醇(TC)、高密度脂蛋白膽固醇(HDL-C)、低密度脂蛋內(nèi)膽固醇(LDL-C)、游離脂肪酸(FFA)及肝功能(ALT、AST)相關(guān)指標(biāo);HE染色觀察脂肪組織病理形態(tài)學(xué)改變;;RT-PCR、Western Blot法檢測(cè)脂肪組織中脂肪分解及成脂分化轉(zhuǎn)錄相關(guān)因子 ATGL、HSL、β3-AR、CEBPα、PPARα、PPARγ 及 SREBP-1 的 mRNA 及蛋白表達(dá)量。結(jié)果在體重變化方面,我們發(fā)現(xiàn)高脂飲食誘導(dǎo)8周時(shí),HFD組小鼠有29只(48.3%);12周時(shí),HFD組小鼠共有47只肥胖模型成功,成模率達(dá)78.3%。治療8周后,模型組小鼠體重平均增加15.71g;正常組小鼠體重平均增加8.315g;丹酚酸B組小鼠體重平均增加0.987g;姜黃素組小鼠體重平均增加2.07g;而陽性藥物二甲雙胍組小鼠體重則平均降低了 5.092g。各治療組與模型組相比,體重增加均明顯低于模型組(二甲雙胍組、丹酚酸B組P0.01;姜黃素組P0.05)。監(jiān)測(cè)各組小鼠體脂率變化發(fā)現(xiàn):在治療第4周時(shí),模型組小鼠體脂率顯著高于正常組(P0.05),而與模型組相比,各治療組小鼠體脂率均有所降低,尤以陽性藥二甲雙胍組小鼠的體脂率較低較為顯著,存在統(tǒng)計(jì)學(xué)差異,而受試藥丹酚酸B、姜黃素組小鼠體脂率雖較模型組有所下降,但無統(tǒng)計(jì)學(xué)差異。在治療第8周時(shí),與模型組相比,各治療組小鼠體脂率均顯著降低(丹酚酸B、姜黃素組P0.01,二甲雙胍組P0.001)。各組小鼠攝食量總體呈現(xiàn)上升趨勢(shì),丹酚酸B、姜黃素在本研究所用的藥物劑量下,均未影響小鼠攝食量。與模型組相比,各治療組小鼠治療第2周后血糖開始下降,在第5周時(shí)血糖值降幅較大,達(dá)到治療期間最低峰值后又出現(xiàn)走高趨勢(shì);而丹酚酸B組至第7、8周時(shí)空腹血糖開始逐漸回落,在給藥第7、8周時(shí),各治療組血糖水平較模型組顯著降低(P0.01)。4周時(shí)OGTT結(jié)果顯示,各治療組曲線下面積(AUC)均小于模型組(P0.01),尤以二甲雙胍組最為顯著。8周時(shí)OGTT曲線下面積顯示,各治療組AUC小于模型組,統(tǒng)計(jì)學(xué)差異顯著(P0.01)。給藥8周后,各治療組均降低了肥胖小鼠血清中TG、TC、LDL-C、FFA的含量(P0.01、P0.05),而血清HDL-C水平較模型組小鼠有所回升(P0.01)。經(jīng)8周治療后,丹酚酸B、姜黃素組血清ALT、AST水平與模型組相比,均出現(xiàn)不同程度降低,具有統(tǒng)計(jì)學(xué)差異(P0.01)。在褐色脂肪組織(BAT)脂解因子的基因表達(dá)方面,與模型組相比,二甲雙胍顯著上調(diào)了 Adiponectin、ATGL、β3-AR的mRNA的表達(dá)量(P0.05);丹酚酸B組顯著上調(diào)了 HSL的mRNA表達(dá)(P0.05)。姜黃素組顯著上調(diào)了 β3-AR的mRNA表達(dá)(P0.05)。BAT在成脂轉(zhuǎn)錄因子的基因表達(dá)方面,丹酚酸B、二甲雙胍組上調(diào)了 C/EBPα、PPARγ的mRNA表達(dá)(P0.05),而姜黃素組雖有上調(diào)趨勢(shì),但無顯著性差異(P0.05)。各治療組對(duì)BAT中PPARα、SREBP-1 mRNA的表達(dá)無明顯影響。在白色脂肪組織(WAT)脂解因子的基因表達(dá)方面,二甲雙胍組中Adiponectin、ATGL及β 3-AR的mRNA的表達(dá)量較模型組相比均顯著上調(diào)(P0.05),姜黃素組顯著上調(diào)HSL、β 3-AR的mRNA表達(dá)(P0.05)。丹酚酸B組雖有上調(diào)ATGL、HSL及β3-ARmRNA的趨勢(shì),但無顯著性差異(P0.05)。在成脂轉(zhuǎn)錄因子的基因表達(dá)方面,與模型組相比,二甲雙胍組、丹酚酸B組均上調(diào)了 C/EBPα的mRNA表達(dá),差異有統(tǒng)計(jì)學(xué)意義(P0.05),各治療組均顯著上調(diào)PPARγ的mRNA表達(dá)(丹酚酸B、姜黃素P0.05;二甲雙胍P0.01)。各治療組對(duì)WAT中PPARα、SREBP-1的mRNA表達(dá)較模型組均無明顯差異。在BAT中相關(guān)脂解蛋白的表達(dá)方面,與模型組相比,各治療組中ATGL蛋白表達(dá)量升高,具有顯著性差異(P0.05)。姜黃素組的HSL蛋白表達(dá)顯著上調(diào)P0.05)。而β3-AR的蛋白表達(dá)各組無明顯差異。在成脂轉(zhuǎn)錄因子的蛋白白表達(dá)方面,各治療組均顯著上調(diào)了 C/EBPα、PPARγ的蛋白表達(dá)(P0.05)。二甲雙胍組、丹酚酸B組顯著增加了 PPARα的蛋白表達(dá)(P0.05),而姜黃素組對(duì)PPARα的蛋白表達(dá)無明顯影響。丹酚酸B組顯著降低SREBP-1的蛋白表達(dá)(P0.01),而姜黃素組及二甲雙胍組雖有降低趨勢(shì),但不具有顯著性差異(P0.05)。在WAT中相關(guān)脂解蛋白表達(dá)方面,與模型組相比,姜黃素組顯著上調(diào)了 HSL的蛋白表達(dá)(P0.05);各治療組中ATGL、β 3-AR的蛋白表達(dá)方面與模型組相比均無明顯差異。在成脂轉(zhuǎn)錄因子的蛋白表達(dá)方面,與模型組相比,各治療組均顯著下調(diào)了PPARy的蛋白表達(dá)(P0.05),各治療組在C/EBPα的蛋白表達(dá)方面,雖有下調(diào)趨勢(shì),但無統(tǒng)計(jì)學(xué)差異(P0.05);此外,丹酚酸B組及二甲雙胍組還顯著降低了 SREBP-1的蛋白表達(dá)(P0.01)。結(jié)論1.降糖消渴顆�；钛鼋M分丹酚酸B、姜黃素可減輕高脂飲食誘導(dǎo)的肥胖C57BL/6J小鼠的體重及體脂含量,降低血清TG、TC、LDL-C及FFA,升高HDL-C,并可減輕肥胖導(dǎo)致的小鼠肝臟損傷,調(diào)節(jié)脂代謝,改善脂代謝紊亂。2.降糖消渴顆粒活血化瘀組分丹酚酸B、姜黃素可降低肥胖C57BL/6J小鼠的空腹血糖,提高胰島素敏感性,減輕肥胖引起的胰島素抵抗,調(diào)節(jié)糖代謝,改善糖代謝紊亂。3.丹酚酸B、姜黃素改善肥胖C57BL/6J小鼠糖脂代謝作用機(jī)制可能通過調(diào)控轉(zhuǎn)錄因子CEBPα、PPARy、SREBP-1參與成脂分化;調(diào)控HSL、β3-AR參與脂肪分解而發(fā)揮作用的。二、降糖消渴顆粒升清活血化瘀組分對(duì)3T3-L1前脂肪細(xì)胞分化及功能的影響目的本部分實(shí)驗(yàn)以不同濃度的丹酚酸B、姜黃素干預(yù)3T3-L1脂肪細(xì)胞,通過對(duì)細(xì)胞形態(tài)、生長增殖、脂質(zhì)含量等細(xì)胞生物學(xué)研究,觀察降糖消渴顆粒活血化瘀組分對(duì)細(xì)胞分化、功能及形態(tài)學(xué)的影響;采用RT-PCR法檢測(cè)其對(duì)3T3-L1前脂肪細(xì)胞脂解及成脂分化轉(zhuǎn)錄因子相關(guān)mRNA表達(dá)的影響,探討降糖消渴顆粒活血化瘀組分影響脂肪細(xì)胞分化和功能的可能作用機(jī)制。方法通過細(xì)胞細(xì)胞增殖與活力檢測(cè)篩選丹酚酸B、姜黃素的安全給藥濃度。將3T3-L1前脂肪細(xì)胞接種于培養(yǎng)板,當(dāng)細(xì)胞生長覆蓋面積達(dá)90%以上時(shí),進(jìn)行誘導(dǎo)分化,8-12天細(xì)胞分化成熟。將分化成熟的3T3-L1脂肪細(xì)胞分為正常對(duì)照組;丹酚酸B組(50μM、75 μ M、100 μ M);姜黃素組(10 μ M、20 μ M、35 μ M)。空白組給予含有等體積DMSO正常培養(yǎng)基進(jìn)行均一化對(duì)照,每組設(shè)4個(gè)復(fù)孔,藥物干預(yù)48h。采用比色法測(cè)定甘油釋放量及葡萄糖消耗量,評(píng)價(jià)藥物干預(yù)后3T3-L1細(xì)胞分化和脂肪分解水平及葡萄糖消耗量:油紅O染色法觀測(cè)細(xì)胞分化及脂滴形態(tài);RT-PCR法檢測(cè)脂肪細(xì)胞中脂肪分解及成脂分化轉(zhuǎn)錄相關(guān)基因ATGL、HSL、β3-AR、CEBP αα、PPARα、PPAR γ及SREBP-1 的 mRNA 表達(dá)。結(jié)果在細(xì)胞細(xì)胞增殖與活力的影響方面,與對(duì)照組相比,丹酚酸B在50、75、100 μM濃度時(shí),其抑制作用并無統(tǒng)計(jì)學(xué)差異,而當(dāng)濃度≥125 μM時(shí),丹酚酸B對(duì)細(xì)胞生長抑制作用具有顯著差異(P0.01)。當(dāng)姜黃素濃度≥50 μM時(shí),姜黃素對(duì)細(xì)胞生長抑制作用有統(tǒng)計(jì)學(xué)意義(P0.01)。并且細(xì)胞出現(xiàn)了成片脫落、死亡。丹酚酸B、姜黃素可促進(jìn)3T3-L1前脂肪細(xì)胞的分化,增加脂質(zhì)堆積。濃度為100μM的丹酚酸B促分化、增加脂質(zhì)堆積的作用最強(qiáng),呈現(xiàn)出一定的量效關(guān)系。不同濃度姜黃素的量效關(guān)系不顯著。在葡萄糖消耗方面,給藥24h時(shí),50 μM、75 μM丹酚酸可顯著增加葡萄糖消耗量(P0.05),而100 μM濃度的丹酚酸B對(duì)葡萄糖消耗的作用不明顯。20μM、35μM濃度的姜黃素也可增加葡萄糖消耗量(P0.05),其中姜黃素的濃度在20 μ M時(shí)最為顯著(P0.01)。干預(yù)48h時(shí),各濃度的丹酚酸B及姜黃素均可增加葡萄糖消耗量(P0.05),而50 μ M、75 u M濃度的丹酚酸B和10 μ M濃度的姜黃素對(duì)葡萄糖消耗的作用最為顯著(P0.01)。在對(duì)3T3-L1脂肪細(xì)胞甘油釋放量的影響方面,丹酚酸B干預(yù)24h時(shí)即可顯著抑制細(xì)胞的甘油釋放量,存在統(tǒng)計(jì)學(xué)差異(P0.01),而各姜黃素組在干預(yù)24h時(shí)與空白組未見明顯差異。干預(yù)48h時(shí),各濃度的丹酚酸及姜黃素均可顯著抑制3T3-L1脂肪細(xì)胞的甘油釋放量,差異具有統(tǒng)計(jì)學(xué)意義(P0.01)。在丹酚酸B對(duì)脂解因子及成脂分化轉(zhuǎn)錄因子mRNA表達(dá)方面:與對(duì)照組相比,各濃度的丹酚酸B顯著下調(diào)了 ATGL、HSL的mRNA表達(dá)量(P0.05);高濃度的丹酚酸B顯著上調(diào)了 C/EBPα的mRNA表達(dá)(P0.05);各濃度的丹酚酸B均明顯上調(diào)了 PPARγ的mRNA表達(dá)(P0.01)。中、高濃度的丹酚酸B上調(diào)了 PPARα的mRNA表達(dá)(P0.05)。在姜黃素對(duì)3T3-L1脂肪細(xì)胞脂解因子的基因表達(dá)方面:與對(duì)照組相比,低濃度的姜黃素下調(diào)ATGL的表達(dá)(P0.05)。中、高濃度的姜黃素可顯著下調(diào)HSL的表達(dá)(P0.01)。高濃度的姜黃素顯著下調(diào)了 C/EBPα的mRNA表達(dá)(P0.05)。中濃度的姜黃素顯著上調(diào)了 PPARγ的mRNA表達(dá)(P0.05)。各濃度組的姜黃素雖出現(xiàn)上調(diào)PPARα mRNA表達(dá)的趨勢(shì),但無顯著性差異。結(jié)論1.丹酚酸B、姜黃素可以促進(jìn)3T3-L1前脂肪細(xì)胞分化,增加脂肪細(xì)胞對(duì)葡萄糖的消耗和利用,改善胰島素抵抗。2.丹酚酸B、姜黃素通過促進(jìn)分化進(jìn)而增強(qiáng)脂肪細(xì)胞功能,抑制脂肪細(xì)胞TG合成及分解、促進(jìn)脂肪酸過氧化,減少FFA釋放,調(diào)節(jié)脂質(zhì)代謝,起到減肥降脂的作用。3.丹酚酸B、姜黃素對(duì)3T3-L1前脂肪細(xì)胞細(xì)胞分化與功能的調(diào)節(jié)作用,可能通過調(diào)控轉(zhuǎn)錄因子CEBP α、PPAR γ及PPARα參與成脂分化;調(diào)控ATGL、HSL參與脂肪分解而發(fā)揮作用。
[Abstract]:First, the effect of Jiangtang Xiaoke Huoxue Huayu component on hyperlipidemic diet induced glucose and lipid metabolism in C57BL/6J mice induced by high fat diet, the effect of high fat diet induced C57BL/6J obese mice model was used to observe the effect of Jiangtang Xiaoke granules in activating blood and removing blood stasis group of salvianolic acid B, curcumin on obesity and fat fat related abnormal glucose and fat metabolism; The fat decomposition and the key transcription factor index of fat differentiation in adipose tissue of obese C57BL/6J mice were detected, and the possible mechanism of regulating the function of adipose tissue and improving the metabolism of fat and fat was further explored. Methods 70 male mice of 6 weeks old were divided into positive body weight randomly. In normal control group (n=10) and high fat group (n=60), normal control group was fed with normal total feed, and high fat group (HFD) mice were fed with high fat diet for.12 weeks, and 20% of the average weight of the normal diet group was obese. The successful standard was to divide the fat mice into model group, metformin group and salvianol randomly. Acid B, curcumin group (n=1 1). Each group was administered by 10g.BW/O.1ml volume. The dose of salvianolic acid B was 100mg/kg.BW/day; the dose of curcumin was 50mg/kg.BW/day; metformin dosage was 75mg/kg.BW/day, the model group and the normal group were given the same amount of deionized water for 8 weeks. The weight, food intake and fasting blood glucose (FB) at the same time per week were monitored at the same time. G), the body fat content and oral glucose tolerance (OGTT) in each group were measured at the end of 4,8. After the experiment, the samples were collected, and the samples were measured in blood, GG (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein (LDL-C), free fatty acid (FFA) and liver function (ALT, AST), and HE staining was used to observe adipose tissue disease. RT-PCR, Western Blot method was used to detect the fat decomposition and lipid differentiation in the adipose tissue, ATGL, HSL, beta 3-AR, CEBP a, PPAR a, PPAR gamma and SREBP-1 mRNA and protein expression. Results in the body weight change, we found 29 (48.3%) in the HFD group at 8 weeks when the high fat diet was induced, and at 12 weeks, the group was small. A total of 47 obesity models were successful in rats. After 8 weeks of 78.3%. treatment, the average weight of the model mice increased by 15.71g, the average weight of the normal group increased by 8.315g, the average weight of the salvianolic acid B mice increased 0.987g, and the weight of the curcumin group increased by 2.07g, while the weight of the metformin group of the positive drug group decreased by 5.092g. on average. The weight increase of each treatment group was significantly lower than that in the model group (metformin group, salvianolic acid B group P0.01, and curcumin group P0.05). The change of body fat rate of the mice in each group was significantly higher than that of the normal group (P0.05) at the fourth week of treatment, but the body fat rate of the mice in the treatment group was lower than that of the model group. The body fat rate of the positive drug metformin group was lower than that of the model group, but the body fat rate of the curcumin group was lower than that of the model group, but the body fat rate of the curcumin group was significantly lower than that of the model group (B of salvianolic acid, curcumin group P0.0) at the eighth week of the treatment. 1, metformin group P0.001). The total intake of food in each group showed an upward trend. The dose of salvianolic acid B, curcumin did not affect the intake of food in mice under this study. Compared with the model group, the blood sugar began to decline after second weeks of treatment in the treatment group, and the blood sugar decreased greatly at fifth weeks, reaching the lowest peak of the period of treatment and then after the treatment. The high trend of walking was found in the time and space of salvianolic acid B and 7,8 week. The blood sugar level of the treatment group was significantly lower than that in the model group (P0.01) at week 7,8 (P0.01). The result showed that the area under the curve of the treatment group (AUC) was less than that of the model group (P0.01), especially under the.8 week OGTT curve of the metformin group. The product showed that the AUC of the treatment group was less than the model group and the statistical difference was significant (P0.01). After 8 weeks of administration, all the treatment groups decreased the content of TG, TC, LDL-C, FFA in the obese mice (P0.01, P0.05), but the serum HDL-C level was higher than that of the model group (P0.01). After 8 weeks of treatment, the salvianolic acid B, the curcumin group's serum ALT, and the model group phase Compared with the model group, metformin significantly up-regulated the expression of mRNA in Adiponectin, ATGL, and beta 3-AR (P0.05), and the salvianolic acid B group significantly up-regulated the HSL mRNA expression (P0.05). The curcumin group was significantly up-regulated, compared with the model group. The mRNA expression of beta 3-AR (P0.05).BAT in the gene expression of lipid transcription factors, the B of salvianolic acid and the mRNA expression of PPAR gamma (P0.05) in the group of C/EBP alpha and PPAR gamma (P0.05), but there is no significant difference in the curcumin group, but there is no significant difference in the expression of PPAR alpha in BAT. The expression of Adiponectin, ATGL and beta 3-AR in the metformin group increased significantly compared with the model group (P0.05). The curcumin group significantly up-regulated HSL and the mRNA expression of beta 3-AR (P0.05). Although the B group of salvianolic acid has the trend of up ATGL, HSL and beta, there is no significant difference in lipid transcription factors. Compared with the model group, the mRNA expression of C/EBP a was up regulated in the metformin group and the salvianolic acid B group compared with the model group, and the difference was statistically significant (P0.05). All the treatment groups significantly increased the mRNA expression of PPAR gamma (salvianolic acid B, curcumin P0.05, and metformin P0.01). The expression of SREBP-1 was no more than that of the model group. There was significant difference in the expression of related lipo protein in BAT. Compared with the model group, the expression of ATGL protein in each treatment group increased significantly (P0.05). The expression of HSL protein in the curcumin group was significantly up to P0.05. There was no significant difference in the protein expression of the beta 3-AR. In the white expression of the lipid transcription factor, the treatment groups were in the treatment group. The protein expression of C/EBP alpha and PPAR gamma (P0.05) was significantly increased. The protein expression of PPAR a was significantly increased in the group of metformin B (P0.05), while the curcumin group had no obvious effect on the expression of PPAR alpha protein. The salvianolic acid B group significantly reduced the SREBP-1 protein expression (P0.01), while the curcumin group and the metformin group had a decreasing trend, but they did not have a tendency to decrease the protein expression (P0.01), but not the curcumin group and the metformin group. There was significant difference (P0.05). Compared with the model group, the curcumin group significantly increased the protein expression of HSL (P0.05) in the expression of related lipo protein in WAT, and there was no significant difference in the expression of ATGL and beta 3-AR from the model group in the treatment groups. The protein expression of PPARy was significantly reduced (P0.05). Although there was a downward trend in the protein expression of C/EBP alpha in the treatment groups, there was no statistical difference (P0.05). In addition, the protein expression of SREBP-1 was significantly reduced in the B and metformin groups of salvianolic acid (P0.01). Conclusion the 1. hypoglycemic and Xiaoke granules are divided into salvianolic acid B, and curcumin can be reduced. The weight and body fat content of obese C57BL/6J mice induced by light and high fat diet, reduce serum TG, TC, LDL-C and FFA, increase HDL-C, and reduce the liver damage caused by obesity, regulate lipid metabolism, improve lipid metabolism disorder,.2. hypoglycemic and Xiaoke granules in the group of salvianolic acid B, and curcumin can reduce the fasting blood sugar of obese C57BL/6J mice. Improve insulin sensitivity, reduce obesity induced insulin resistance, regulate glucose metabolism, improve glycometabolism.3. salvianolic acid B, curcumin improves the glycolipid metabolism mechanism of obese C57BL/6J mice by regulating transcription factor CEBP alpha, PPARy, SREBP-1 to participate in lipid differentiation; regulation of HSL, beta 3-AR involved in fat decomposition and play a role. Two, two, The effect of Jiangtang Xiaoke Granule on the differentiation and function of 3T3-L1 preadipocyte by promoting blood circulation and removing blood stasis in the experiment with different concentrations of salvianolic acid B, curcumin intervention in 3T3-L1 adipocytes, cell biology, cell morphology, growth and proliferation, lipid content and so on, to observe the blood circulation and blood stasis components of hypoglycemic and Xiaoke granules to the cells The effect of differentiation, function and morphology, RT-PCR method was used to detect the effect of RT-PCR on the lipogenesis of preadipocytes and the expression of lipogenic differentiation transcription factor related to 3T3-L1, and to explore the possible mechanism of influencing the differentiation and function of adipocyte by activating blood and removing blood stasis components of Jiangtang Xiaoke granules. B, the safe dosage of curcumin. Inoculating 3T3-L1 preadipocytes into the culture plate. When the cell growth coverage area was over 90%, the cells were induced to differentiate and the cells differentiated and mature in 8-12 days. The mature 3T3-L1 adipocytes were divided into the normal control group, the salvianolic acid B group (50 mu M, 75 mu M, 100 mu M), and the curcumin group (10 mu M, 20 u M, 35 micron M). The blank group was treated with equal volume DMSO normal medium for homogenization control. Each group had 4 compound holes. Drug intervention 48h. used colorimetric method to determine the amount of glycerol release and glucose consumption. The differentiation and adipose level of 3T3-L1 cells and glucose consumption after drug intervention were evaluated: oil red O staining method was used to observe the cell differentiation and lipid droplet morphology; RT ATGL, HSL, beta 3-AR, CEBP alpha, PPAR a, PPAR gamma and mRNA expression in fat cells were detected by -PCR, and the inhibitory effect of salvianolic B in the concentration of 50,75100 micron was not statistically significant compared with the control group. At 125 mu M, the inhibitory effect of salvianolic acid B on cell growth was significantly different (P0.01). When the concentration of curcumin was more than 50 M, curcumin had significant inhibitory effect on cell growth (P0.01). And the cells appeared to fall off and died. Salvianolic acid B, curcumin can promote the differentiation of 3T3-L1 preadipocytes and increase the accumulation of lipid. The concentration is 100. M of salvianolic acid (B) has the strongest effect on increasing lipid accumulation and showing a certain dose effect relationship. The dose effect relationship of different concentrations of curcumin is not significant. In terms of glucose consumption, 50 mu M, 75 mu M salvianolic acid can significantly increase the glucose consumption (P0.05), while the effect of salvianolic acid B with 100 mu M concentration is not effective on glucose consumption. Obviously.20 mu M, 35 mu M concentration of curcumin can also increase glucose consumption (P0.05), of which the concentration of curcumin at 20 mu is the most significant (P0.01). When intervention 48h, the concentration of salvianolic acid B and curcumin can increase the glucose consumption (P0.05), while 50 mu M, 75 u M consistency and 10 micron concentration of curcumin on glucose consumption The effect was most significant (P0.01). In the effect on the release of glycerol in 3T3-L1 adipocytes, the glycerol release was significantly inhibited by salvianolic acid B intervention in 24h, and there was a statistical difference (P0.01), but there was no significant difference between the curcumin group and the blank group during the intervention of 24h. The concentration of salvianolic acid and curcumin at each concentration could be displayed when interfering with 48h. The difference in the release of glycerol in 3T3-L1 adipocytes was statistically significant (P0.01). In salvianolic acid B, the expression of Lipo factor and lipid differentiation transcription factor mRNA: compared with the control group, the concentration of salvianolic acid B significantly lowered ATGL, HSL mRNA expression (P0.05), and high concentration of salvianolic acid B significantly increased C/EBP alpha mRNA table. (P0.05); all concentration of salvianolic acid B significantly up-regulated the mRNA expression of PPAR gamma (P0.01). High concentration of salvianolic acid B up regulation of PPAR alpha mRNA expression (P0.05). Gene expression of curcumin to 3T3-L1 adipocyte lipolytic factor: compared with the control group, low concentration curcumin down ATGL expression (P0.05). High concentration of Jiang Huang The expression of HSL was significantly down regulated (P0.01). The high concentration curcumin significantly lowered the mRNA expression of C/EBP alpha (P0.05). The concentration of curcumin significantly up-regulated the mRNA expression of PPAR gamma (P0.05). Although the curcumin increased the expression of PPAR alpha mRNA in each concentration group, there was no significant difference. Conclusion 1. salvianolic acid B, curcumin can promote 3T3-. L1 differentiation of preadipocytes, increased consumption and utilization of glucose by adipocytes, improved insulin resistance,.2. salvianolic acid B, and curcumin promoted
【學(xué)位授予單位】：北京中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：R285.5

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