胰岛素诱导基因（Insig2）在调控脂肪细胞分化与脂肪合成中作用的研究
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摘要
第一部分葡萄糖浓度对3T3-L1细胞分化及insig-1和-2 mRNA表达的影响
目的胰岛素诱导基因(insig)是近年来发现的新基因，在脂质合成与脂肪细胞的分化中发挥着重要的调控作用。观察不同葡萄糖浓度对3T3-L1细胞分化及insig-1，-2 mRNA表达的影响，探讨insig在脂肪细胞分化与脂肪代谢中的作用。
方法将3T3-L1细胞分别在不同葡萄糖浓度(5.5mol／L和25mol／L)培养液中诱导分化，用油红“O”染色、RT-PCR和原位杂交技术检测脂肪细胞的分化、insig-1，-2 mRNA及脂肪细胞脂肪酸结合蛋白(AP_2) mRNA的表达。
结果：随着3T3-L1细胞的分化，insig-1，-2 mRNA、AP_2 mRNA表达逐渐上调；但低糖诱导组的细胞分化程度显著低于高糖诱导组；低糖诱导组的insig-1，-2 mRNA表达较高糖诱导组明显增高(P＜0.05)，而AP_2 mRNA表达较高糖诱导组明显降低(P＜0.05)。
结论在3T3-L1细胞分化过程心中，insig基因表达逐渐上调，低糖时insig的表达上调更明显，此可能与低糖时脂肪细胞分化及脂质沉积相对受抑制有关。
第二部分insig-2过表达对3T3-L1前脂肪细胞分化与相关基因的影响
第一章EGFP-C_3-insig-2和pcDNA3.1(+)-insig-2真核表达质粒的构建及insig-2的亚细胞定位
目的构建EGFP-C3-insig-2和pcDNA3.1(+)-insig-2融合基因真核表达质粒，探讨insig-2基因的亚细胞定位与表达，以及稳定转染的3T3-L1细胞insig-2过表达对前脂肪细胞分化的影响。
方法采用RT-PCR法从3T3-L1细胞获取小鼠的全长insig-2基因，经T载体克隆、鉴定后，分别克隆入真核表达载体pEGFP-C_3及pcDNA3.1(+)中。酶切、测序对插入片段进行分析和鉴定后，用脂质体转染法将带绿色荧光蛋白的真核表达载体pEGFP-C_3瞬时转入3T3-L1细胞，通过荧光显微镜、RT-PCR检测其在3T3-L1细胞中的表达、定位及下游基因AP_2 mRNA的变化。
结果：酶切、测序鉴定所插入片段的大小和基因序列准确无误，成功构建了重组质粒pEGFP-C_3-insig-2和pCDNA3.1(+)-insig-2融合基因真核表达载体。pEGFP-C_3-insig-2真核表达载体瞬时转染3T3-L1细胞后，该融合蛋白在转染的细胞胞浆中表达；insig-2基因的转录明显增强，而其下游基因AP2 mRNA表达明显下调。
结论：insig-2蛋白在3T3-L1细胞中定位于胞浆，过表达时影响其脂肪代谢。
第二章insig-2基因稳定转染对3T3-L1细胞分化和脂质合成的影响
目的：研究pCDNA3.1(+)-insig-2稳定转染3T3-L1细胞后insig-2基因过表达在3T3-L1脂肪细胞分化和脂肪形成中的作用。
方法：用脂质体转染法将重组质粒pCDNA3.1(+)-insig-2稳定转染3T3-L1细胞。转染后，分别用流式细胞仪检测pCDNA3.1(+)-insig-2转染组、pCDNA3.1(+)空质粒组及未转染的3T3-L1细胞(空白对照组)的细胞周期与凋亡率；用油红“O”染色鉴定分析细胞的诱导分化；用半定量RT-PCR测定各组3T3-L1前脂肪细胞及分化过程中insig-2、insig-1、固醇调节元件结合蛋白(sterol regulatory element binding proteins SREBPs)、SREBP裂解活性蛋白(cleavage-actiwating protein SCAP)、脂肪酸合成酶(FAS)、AP2基因的mRNA表达；用免疫组织化学法检测细胞转染后的insig-2蛋白表达。
结果：半定量RT-PCR检测和免疫组化分析结果显示，insig-2 mRNA及蛋白表达较转染前及空质粒转染组、空白对照组均显著增加，提示pCDNA3.1(+)-insig-2成功转染的3T3-L1细胞呈高效表达，但对其前体细胞的细胞周期及凋亡率无明显影响；诱导分化后的第6和12天，pCDNA3.1(+)-insig-2转染组的成熟脂肪细胞明显少于两个对照组，且分化过程中三组细胞生长曲线无明显差异。转染后的3T3-L1前脂肪细胞insig-2相关基因insig-1、FAS、AP2、SREBPlc mRNA均表达下调；随着细胞的分化，上述基因mRNA表达均上调，但pCDNA3.1(+)-insig-2转染组的insig-2较其他两个对照组显著上调，而FAS和AP2 mRNA明显下调。
结论：转染后insig-2高表达抑制3T3-L1前脂肪细胞的分化及脂质合成相关基因表达。
第三章insig-2基因过表达对3T3-L1脂肪细胞脂联素表达的影响
目的：脂联素对糖代谢、脂代谢和能量平衡的调控有重要作用。观察pCDNA3.1(+)-insig-2转染后3T3-L1前脂肪细胞脂联素mRNA表达及其分化过程中的脂联素分泌，探讨insig基因过表达对脂肪细胞脂联素分泌的影响。
方法：以转染pCDNA3.1(+)及未转染的3T3-L1细胞为对照组，半定量RT-PCR检测pCDNA3.1(+)-insig-2转染后24 h和72h 3T3-L1前脂肪细胞的脂联素mRNA表达。用ELISA法测定转染后3T3-L1前脂肪细胞及其在诱导分化过程中的脂联素动态变化。
结果：3T3-L1前脂肪细胞pCDNA3.1(+)-insig-2转染后，脂联素mRNA的表达量明显降低；3T3-L1前脂肪细胞的脂联素分泌较对照组明显降低(P＜0.05)；在诱导分化的第0～4天，三组的脂联素下降无明显差异，但此后其分泌逐渐升高，转染组较其他两个对照组明显降低(P＜0.05)。
结论：转染后insig-2过表达抑制3T3-L1脂肪细胞脂联素mRNA和蛋白的表达。
Part 1 Effects of different concentration of glucose on the expression ofinsig-1, -2 and the differentiation in 3T3-L1 preadipocytes
Objective: Insulin-induced gene 1 (insig-1), plays an important regulatory role in lipogenesis andadipocytes differentiation. The effects of insig-lon adipocytes differentiation and lipogenesis, andthe expression of insig-1, -2 during 3T3-L1 preadipocyte differentiation under differentconcentration of glucose were investigated.
Methods: 3T3-L1 preadipocytes differentiation was induced with different concentration of glucose(5.5 and 25mol/L). Cell differentiation was observed by oil red O staining, RT-PCR and in situhybridization were used to detect the expressions of insig-1,-2, and fatty acid binding protein(AP_2)mRNA
Results: With 3T3-L1 preadipocytes differentiation, the expression of insig-1,2 and AP_2 mRNAwas gradually increased. However, the differentiated cell extent under low glucose concentrationwas significantly lower than that under high glucose concentration. A higher expression of insig-1,-2 mRNA with a lower expression of AP_2 mRNA was observed under low glucoseconcentration (P＜0.05).
Conclusion: During 3T3-L1 preadipocytes differentiation, the expression of insig was graduallyincreased. In low glucose condition, much higher expression of insig was probably related to ainhibition of cell differentiation and lipidoses
Part 2 Effects of insig-2 overexpression on 3T3-L1 preadipocytesdifferentiation and the expression of related genes
Chapter 1Construction of eukaryotic expression vectors EGFP-C3-insig-2,pcDNA3.1(+)-Insig-2, and the subcellular localization of insig-2 in 3T3-L1preadipocytes
Objective: To construct the eukaryotic expression vectors EGFP-C3-insig-2, pcDNA3.1(+)-Insig-2and to subcellularly localize insig-2 protein in 3T3-L1 preadipocytes.
Method: The open reading frame (ORF) of insig-2 was obtained from 3T3-L1 pre- adipocytes byRT-PCR. The fragment encoding for insig-2 gene was inserted into pUCm-T vector, sequenced, andidentified by PCR. pUCm-T-insig-2 was digested by two restrictive enzymes and insig-2 wassubcloned into eukaryotic expressing vector EGFP-C_3 and pcDNA3.1(+)-Insig-2. The recombinedplasmid EGFP-C3-insig-2 was transfected transiently into 3T3-L1 preadipocytes withLipofeetamineTM 2000 Transfection Reagent. Green fluorescence protein expression andlocalization of the plasmid were determined by fluorescence microscopy, and the expression ofdown-stream gene AP_2 was detected by RT-PCR.
Results: Eukaryotic expression vectors EGFP-C3-insig-2 and pcDNA3.1(+)-Insig-2 wasconstructed successfully. After EGFP-C3-insig-2 transfected into 3T3-L1 preadipocytes,insig-2protein was found to localize in cytoplasm other than nucleus. RT-PCR showed that the amount ofinsig-2 transcription was significantly enhanced with a lowered expression of its down-stream geneAP2.
Conclusion: Overexpression of insig-2 could produce an impact on cell lipid metabolism during3T3-L1 preadipocyte differentiation.
Chapter 2Effects of insig-2 stable expression on 3T3-L1 preadipocytes differentiation andlipogenesis
Objective: To study the effects of insig-2 stable expression on 3T3-L1 preadipocytes differentiationand lipogenesis.
Methods: The recombinant plasmid pcDNA3.1(+)-insig-2 and control plasmid pcDNA3.1(+) weretransfected into 3T3-L1 preadipocytes with LipofeetamineTM 2000 Transfection Reagent and thepositive clones were screened by G418. Cell cycles and apoptosis index in pcDNA3.1(+)-insig-2,pcDNA3.1(+) and non-transfected cells were analyzed by Flow Cytometry(FCM). Celldifferentiation and cell growth were identified using oil red O staining and cell growthcurves, respectively. Semi-quantitative RT-PCR was performed to detect the expression of insig-1,insig-2, sterol regulatory element binding proteins (SREBPs), cleavage-activating protein (SCAP),fatty acid synthetase (FAS), and fatty acid binding protein (AP2) in 3T3-L1 pre adipocytes anddifferentiation-induced adipocytes. The expression of insig-2 protein was determined byimmunohistochemistry.
Results: Semi-quantitative RT-PCR and immunohistochemistry showed that the expression ofinsig-2 in 3T3-L1 preadipocytes transfected with insig-2 was significantly increased than thattransfected with blank pcDNA3.1(+) vector and none-transfected cells, suggesting thatpcDNA3.1(+)-insig-2 was transfected in 3T3-L1 preadipocytes successfully with high-efficientexpression. There was no remarkable effect on 3T3-L1 preadipocytes cell cycle and apoptosis indexwith insig-2 gene stably expression. After 6 and 12 days of induction, it was found that the maturedadipocytes in transfected pcDNA3.1(+)-insig-2 were showed to have bright red fat droplets with nosignificant changes of the cell growth curve. After transfected with pcDNA3.1(+)-insig-2, theexpression of insig-2 related genes including insig-1, FAS, AP2, and SREBP1c was down-regulated,but with up-expressions during cell differentiation. However, the expression of insig-2 increasedrapidly than that of FAS and AP2.
Conclusion: Overexpression of insig-2 could inhibit 3T3-L1 preadipocytes differentiation intomature adipocytes and suppress the expression of genes related to lipogenesis.
Chapter 3Effects of insig-2 stable expression on adiponectin secretion in 3T3-L1preadipocytes
Objective: Adiponectin plays an important role in the regulation of glucose and lipid metabolismand energy balance. To approach the influence of overexpression of insig-2 on adiponectin secretionin 3T3-L1 adipocytes, the expression of adiponectin mRNA and the adiponectin secretion duringpreadipocytes differentiation were observed.
Methods: After 24 and 72 h of transfection, semi-quantitative RT-PCR was performed to detect theexpression of adiponectin mRNA in transfected with cDNA3.1(+)-insig-2) and transfected withblank pcDNA3.1(+) vector or non-transfected cells. Adiponectin was detected by ELISA.
Results: RT-PCR showed that the expression of adiponectin mRNA was decreased significantlyafter insig-2 gene stable expression in 3T3-L1 preadipocytes with a rather low level of adiponectinsecretion in the culture medium (P＜0.05). It was constently low during 0～4 days of induction, thengradually increased, but was still significant lower in transfected with cDNA3.1(+)-insig-2) cells(P＜0.05).
Conclusion: Overexpression of insig-2 could suppressed the expression of adiponectin mRNA andprotein in 3T3-L1 preadipocytes.
目的胰岛素诱导基因(insig)是近年来发现的新基因，在脂质合成与脂肪细胞的分化中发挥着重要的调控作用。观察不同葡萄糖浓度对3T3-L1细胞分化及insig-1，-2 mRNA表达的影响，探讨insig在脂肪细胞分化与脂肪代谢中的作用。
方法将3T3-L1细胞分别在不同葡萄糖浓度(5.5mol／L和25mol／L)培养液中诱导分化，用油红“O”染色、RT-PCR和原位杂交技术检测脂肪细胞的分化、insig-1，-2 mRNA及脂肪细胞脂肪酸结合蛋白(AP_2) mRNA的表达。
结果：随着3T3-L1细胞的分化，insig-1，-2 mRNA、AP_2 mRNA表达逐渐上调；但低糖诱导组的细胞分化程度显著低于高糖诱导组；低糖诱导组的insig-1，-2 mRNA表达较高糖诱导组明显增高(P＜0.05)，而AP_2 mRNA表达较高糖诱导组明显降低(P＜0.05)。
结论在3T3-L1细胞分化过程心中，insig基因表达逐渐上调，低糖时insig的表达上调更明显，此可能与低糖时脂肪细胞分化及脂质沉积相对受抑制有关。
第二部分insig-2过表达对3T3-L1前脂肪细胞分化与相关基因的影响
第一章EGFP-C_3-insig-2和pcDNA3.1(+)-insig-2真核表达质粒的构建及insig-2的亚细胞定位
目的构建EGFP-C3-insig-2和pcDNA3.1(+)-insig-2融合基因真核表达质粒，探讨insig-2基因的亚细胞定位与表达，以及稳定转染的3T3-L1细胞insig-2过表达对前脂肪细胞分化的影响。
方法采用RT-PCR法从3T3-L1细胞获取小鼠的全长insig-2基因，经T载体克隆、鉴定后，分别克隆入真核表达载体pEGFP-C_3及pcDNA3.1(+)中。酶切、测序对插入片段进行分析和鉴定后，用脂质体转染法将带绿色荧光蛋白的真核表达载体pEGFP-C_3瞬时转入3T3-L1细胞，通过荧光显微镜、RT-PCR检测其在3T3-L1细胞中的表达、定位及下游基因AP_2 mRNA的变化。
结果：酶切、测序鉴定所插入片段的大小和基因序列准确无误，成功构建了重组质粒pEGFP-C_3-insig-2和pCDNA3.1(+)-insig-2融合基因真核表达载体。pEGFP-C_3-insig-2真核表达载体瞬时转染3T3-L1细胞后，该融合蛋白在转染的细胞胞浆中表达；insig-2基因的转录明显增强，而其下游基因AP2 mRNA表达明显下调。
结论：insig-2蛋白在3T3-L1细胞中定位于胞浆，过表达时影响其脂肪代谢。
第二章insig-2基因稳定转染对3T3-L1细胞分化和脂质合成的影响
目的：研究pCDNA3.1(+)-insig-2稳定转染3T3-L1细胞后insig-2基因过表达在3T3-L1脂肪细胞分化和脂肪形成中的作用。
方法：用脂质体转染法将重组质粒pCDNA3.1(+)-insig-2稳定转染3T3-L1细胞。转染后，分别用流式细胞仪检测pCDNA3.1(+)-insig-2转染组、pCDNA3.1(+)空质粒组及未转染的3T3-L1细胞(空白对照组)的细胞周期与凋亡率；用油红“O”染色鉴定分析细胞的诱导分化；用半定量RT-PCR测定各组3T3-L1前脂肪细胞及分化过程中insig-2、insig-1、固醇调节元件结合蛋白(sterol regulatory element binding proteins SREBPs)、SREBP裂解活性蛋白(cleavage-actiwating protein SCAP)、脂肪酸合成酶(FAS)、AP2基因的mRNA表达；用免疫组织化学法检测细胞转染后的insig-2蛋白表达。
结果：半定量RT-PCR检测和免疫组化分析结果显示，insig-2 mRNA及蛋白表达较转染前及空质粒转染组、空白对照组均显著增加，提示pCDNA3.1(+)-insig-2成功转染的3T3-L1细胞呈高效表达，但对其前体细胞的细胞周期及凋亡率无明显影响；诱导分化后的第6和12天，pCDNA3.1(+)-insig-2转染组的成熟脂肪细胞明显少于两个对照组，且分化过程中三组细胞生长曲线无明显差异。转染后的3T3-L1前脂肪细胞insig-2相关基因insig-1、FAS、AP2、SREBPlc mRNA均表达下调；随着细胞的分化，上述基因mRNA表达均上调，但pCDNA3.1(+)-insig-2转染组的insig-2较其他两个对照组显著上调，而FAS和AP2 mRNA明显下调。
结论：转染后insig-2高表达抑制3T3-L1前脂肪细胞的分化及脂质合成相关基因表达。
第三章insig-2基因过表达对3T3-L1脂肪细胞脂联素表达的影响
目的：脂联素对糖代谢、脂代谢和能量平衡的调控有重要作用。观察pCDNA3.1(+)-insig-2转染后3T3-L1前脂肪细胞脂联素mRNA表达及其分化过程中的脂联素分泌，探讨insig基因过表达对脂肪细胞脂联素分泌的影响。
方法：以转染pCDNA3.1(+)及未转染的3T3-L1细胞为对照组，半定量RT-PCR检测pCDNA3.1(+)-insig-2转染后24 h和72h 3T3-L1前脂肪细胞的脂联素mRNA表达。用ELISA法测定转染后3T3-L1前脂肪细胞及其在诱导分化过程中的脂联素动态变化。
结果：3T3-L1前脂肪细胞pCDNA3.1(+)-insig-2转染后，脂联素mRNA的表达量明显降低；3T3-L1前脂肪细胞的脂联素分泌较对照组明显降低(P＜0.05)；在诱导分化的第0～4天，三组的脂联素下降无明显差异，但此后其分泌逐渐升高，转染组较其他两个对照组明显降低(P＜0.05)。
结论：转染后insig-2过表达抑制3T3-L1脂肪细胞脂联素mRNA和蛋白的表达。
Part 1 Effects of different concentration of glucose on the expression ofinsig-1, -2 and the differentiation in 3T3-L1 preadipocytes
Objective: Insulin-induced gene 1 (insig-1), plays an important regulatory role in lipogenesis andadipocytes differentiation. The effects of insig-lon adipocytes differentiation and lipogenesis, andthe expression of insig-1, -2 during 3T3-L1 preadipocyte differentiation under differentconcentration of glucose were investigated.
Methods: 3T3-L1 preadipocytes differentiation was induced with different concentration of glucose(5.5 and 25mol/L). Cell differentiation was observed by oil red O staining, RT-PCR and in situhybridization were used to detect the expressions of insig-1,-2, and fatty acid binding protein(AP_2)mRNA
Results: With 3T3-L1 preadipocytes differentiation, the expression of insig-1,2 and AP_2 mRNAwas gradually increased. However, the differentiated cell extent under low glucose concentrationwas significantly lower than that under high glucose concentration. A higher expression of insig-1,-2 mRNA with a lower expression of AP_2 mRNA was observed under low glucoseconcentration (P＜0.05).
Conclusion: During 3T3-L1 preadipocytes differentiation, the expression of insig was graduallyincreased. In low glucose condition, much higher expression of insig was probably related to ainhibition of cell differentiation and lipidoses
Part 2 Effects of insig-2 overexpression on 3T3-L1 preadipocytesdifferentiation and the expression of related genes
Chapter 1Construction of eukaryotic expression vectors EGFP-C3-insig-2,pcDNA3.1(+)-Insig-2, and the subcellular localization of insig-2 in 3T3-L1preadipocytes
Objective: To construct the eukaryotic expression vectors EGFP-C3-insig-2, pcDNA3.1(+)-Insig-2and to subcellularly localize insig-2 protein in 3T3-L1 preadipocytes.
Method: The open reading frame (ORF) of insig-2 was obtained from 3T3-L1 pre- adipocytes byRT-PCR. The fragment encoding for insig-2 gene was inserted into pUCm-T vector, sequenced, andidentified by PCR. pUCm-T-insig-2 was digested by two restrictive enzymes and insig-2 wassubcloned into eukaryotic expressing vector EGFP-C_3 and pcDNA3.1(+)-Insig-2. The recombinedplasmid EGFP-C3-insig-2 was transfected transiently into 3T3-L1 preadipocytes withLipofeetamineTM 2000 Transfection Reagent. Green fluorescence protein expression andlocalization of the plasmid were determined by fluorescence microscopy, and the expression ofdown-stream gene AP_2 was detected by RT-PCR.
Results: Eukaryotic expression vectors EGFP-C3-insig-2 and pcDNA3.1(+)-Insig-2 wasconstructed successfully. After EGFP-C3-insig-2 transfected into 3T3-L1 preadipocytes,insig-2protein was found to localize in cytoplasm other than nucleus. RT-PCR showed that the amount ofinsig-2 transcription was significantly enhanced with a lowered expression of its down-stream geneAP2.
Conclusion: Overexpression of insig-2 could produce an impact on cell lipid metabolism during3T3-L1 preadipocyte differentiation.
Chapter 2Effects of insig-2 stable expression on 3T3-L1 preadipocytes differentiation andlipogenesis
Objective: To study the effects of insig-2 stable expression on 3T3-L1 preadipocytes differentiationand lipogenesis.
Methods: The recombinant plasmid pcDNA3.1(+)-insig-2 and control plasmid pcDNA3.1(+) weretransfected into 3T3-L1 preadipocytes with LipofeetamineTM 2000 Transfection Reagent and thepositive clones were screened by G418. Cell cycles and apoptosis index in pcDNA3.1(+)-insig-2,pcDNA3.1(+) and non-transfected cells were analyzed by Flow Cytometry(FCM). Celldifferentiation and cell growth were identified using oil red O staining and cell growthcurves, respectively. Semi-quantitative RT-PCR was performed to detect the expression of insig-1,insig-2, sterol regulatory element binding proteins (SREBPs), cleavage-activating protein (SCAP),fatty acid synthetase (FAS), and fatty acid binding protein (AP2) in 3T3-L1 pre adipocytes anddifferentiation-induced adipocytes. The expression of insig-2 protein was determined byimmunohistochemistry.
Results: Semi-quantitative RT-PCR and immunohistochemistry showed that the expression ofinsig-2 in 3T3-L1 preadipocytes transfected with insig-2 was significantly increased than thattransfected with blank pcDNA3.1(+) vector and none-transfected cells, suggesting thatpcDNA3.1(+)-insig-2 was transfected in 3T3-L1 preadipocytes successfully with high-efficientexpression. There was no remarkable effect on 3T3-L1 preadipocytes cell cycle and apoptosis indexwith insig-2 gene stably expression. After 6 and 12 days of induction, it was found that the maturedadipocytes in transfected pcDNA3.1(+)-insig-2 were showed to have bright red fat droplets with nosignificant changes of the cell growth curve. After transfected with pcDNA3.1(+)-insig-2, theexpression of insig-2 related genes including insig-1, FAS, AP2, and SREBP1c was down-regulated,but with up-expressions during cell differentiation. However, the expression of insig-2 increasedrapidly than that of FAS and AP2.
Conclusion: Overexpression of insig-2 could inhibit 3T3-L1 preadipocytes differentiation intomature adipocytes and suppress the expression of genes related to lipogenesis.
Chapter 3Effects of insig-2 stable expression on adiponectin secretion in 3T3-L1preadipocytes
Objective: Adiponectin plays an important role in the regulation of glucose and lipid metabolismand energy balance. To approach the influence of overexpression of insig-2 on adiponectin secretionin 3T3-L1 adipocytes, the expression of adiponectin mRNA and the adiponectin secretion duringpreadipocytes differentiation were observed.
Methods: After 24 and 72 h of transfection, semi-quantitative RT-PCR was performed to detect theexpression of adiponectin mRNA in transfected with cDNA3.1(+)-insig-2) and transfected withblank pcDNA3.1(+) vector or non-transfected cells. Adiponectin was detected by ELISA.
Results: RT-PCR showed that the expression of adiponectin mRNA was decreased significantlyafter insig-2 gene stable expression in 3T3-L1 preadipocytes with a rather low level of adiponectinsecretion in the culture medium (P＜0.05). It was constently low during 0～4 days of induction, thengradually increased, but was still significant lower in transfected with cDNA3.1(+)-insig-2) cells(P＜0.05).
Conclusion: Overexpression of insig-2 could suppressed the expression of adiponectin mRNA andprotein in 3T3-L1 preadipocytes.
引文
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