小檗碱对高胆固醇血症的影响可能涉及肝脏Insig-2基因表达
摘要
目的:
研究小檗碱(berberine,Ber)对胆固醇代谢的影响,并探讨其可能的作用机制。
方法:
1. SD大鼠40只随机分为普食组(normal diet,ND),高脂饲料组(high fat diet,HFD),高脂饲料+洛伐他汀组(lovastatin,LD),高脂饲料+低剂量小檗碱干预组(berberine low dose diet,BLD),高脂饲料+高剂量小檗碱干预组(berberine high dose diet,BHD)。饲养18周后测定大鼠血中胆固醇(total cholesterol,TC)、高密度脂蛋白胆固醇(high densitylipoprotein cholesterol ,HDL-C)、低密度脂蛋白胆固醇(1ow densitylipoprotein cholesterol ,LDL-C)等血生化指标的含量。RT-PCR, Western blot分别检测各组大鼠肝脏胰岛素诱导基因2(Insulin-induced gene 2,Insig-2)mRNA和蛋白表达变化。
2.RT-PCR,Western blot检测不同浓度(1、3、10、30、100μmol/L)Ber处理24 h后及3μmol/L Ber处理不同时间(0、12、24、48、72h)后HepG2细胞中Insig-2和VDR基因表达,并进行相关性分析。
结果:
1.Ber显著降低高脂模型大鼠的TC和LDL-C水平,同时显著升高大鼠血清HDL-C。
2.低剂量Ber能显著增加高脂模型大鼠肝脏Insig-2基因mRNA和蛋白表达水平;高剂量Ber对Insig-2基因mRNA和蛋白表达影响不明显。
3.Ber对HepG2细胞Insig-2和VDR基因表达呈双向调节作用:≤3μmol/L时呈浓度依赖性上调,>3μmol/L表达值则下降,两者的变化具有相关性(r=0.753,p<0.01);时-效关系研究显示,3μmol/L Ber在48 h作用时间内呈时间依赖性上调Insig-2和VDR基因的表达,两者的变化具有相关性(r=0.922,p<0.01)。
结论:
Ber具有明显调血脂作用,其机制可能与其上调VDR的表达,进而上调Insig-2基因表达,从而抑制肝脏中胆固醇的合成代谢有关。
Objective:
To investigate the effects of berberine (Ber) on cholesterol metabolism and its mechanism。
Methods:
1. SD rats were randomly divided into 5 groups: normal diet + pretreatment of solvent (ND); high-fat and high-cholesterol diet + pretreatment of solvent (HFD); high-fat and high-cholesterol diet + pretreatments of either lovastatin(LD) orBer low dose (BLD) and Ber high dose (BHD). The levels of total cholesterol(TC),low density lipoprotein cholesterol (LDL-C) , and high density lipoprotein cholesterol(HDL-C) in blood were detected eighteen weeks after rats were fed with high-fat and high-cholesterol diet and and pretreated with either solvent or drugs. The mRNA and protein expressions of Insulin-induced gene 2(Insig-2) in rat liver were quantified by RT-PCR and Western blot。
2. HepG2 cells were incubated with Ber at different concentrations (1, 3, 10, 30, 100μmol/L), or incubated with 3μmol/L Ber for various times (0, 12, 24, 48, 72 h). The gene expressions of Insig-2 and VDR from HepG2 cells treated by Ber were quantified by RT-PCR and Western blot.
Results:
1. The blood levels of TC and LDL-C were significantly lower in Ber groups than those in HFD, except HDL-C level that was shown to clearly increase after rats were pretreatment with Ber.
2.Low dose Ber pretreatment significantly increased Insig-2 gene mRNA and protein expressions from rat liver but high dose Ber pretreatment had no significantly effects on Insig-2 gene expression.
3.24 h after exposure to gradient concentrations of Ber, the gene expressions of Insig-2 and VDR in HepG2 cells showed a dualistic changes:the maximum values occurred at 3μmol/L, then the values declined gradually. The pretreatment with 3μmol/L Ber for different time made the gene expressions of Insig-2 and VDR significantly increased with the peak value at 48 h;the relative coefficient between them was 0.922 (p<0.01).
Conclusion:
Ber can significantly decrease the levels of hyperlidemia, the mechanism of which may be related to elevating expressions of Insig-2 and VDR genes in liver. As a result the cholesterol synthesis of liver was inhibited.
研究小檗碱(berberine,Ber)对胆固醇代谢的影响,并探讨其可能的作用机制。
方法:
1. SD大鼠40只随机分为普食组(normal diet,ND),高脂饲料组(high fat diet,HFD),高脂饲料+洛伐他汀组(lovastatin,LD),高脂饲料+低剂量小檗碱干预组(berberine low dose diet,BLD),高脂饲料+高剂量小檗碱干预组(berberine high dose diet,BHD)。饲养18周后测定大鼠血中胆固醇(total cholesterol,TC)、高密度脂蛋白胆固醇(high densitylipoprotein cholesterol ,HDL-C)、低密度脂蛋白胆固醇(1ow densitylipoprotein cholesterol ,LDL-C)等血生化指标的含量。RT-PCR, Western blot分别检测各组大鼠肝脏胰岛素诱导基因2(Insulin-induced gene 2,Insig-2)mRNA和蛋白表达变化。
2.RT-PCR,Western blot检测不同浓度(1、3、10、30、100μmol/L)Ber处理24 h后及3μmol/L Ber处理不同时间(0、12、24、48、72h)后HepG2细胞中Insig-2和VDR基因表达,并进行相关性分析。
结果:
1.Ber显著降低高脂模型大鼠的TC和LDL-C水平,同时显著升高大鼠血清HDL-C。
2.低剂量Ber能显著增加高脂模型大鼠肝脏Insig-2基因mRNA和蛋白表达水平;高剂量Ber对Insig-2基因mRNA和蛋白表达影响不明显。
3.Ber对HepG2细胞Insig-2和VDR基因表达呈双向调节作用:≤3μmol/L时呈浓度依赖性上调,>3μmol/L表达值则下降,两者的变化具有相关性(r=0.753,p<0.01);时-效关系研究显示,3μmol/L Ber在48 h作用时间内呈时间依赖性上调Insig-2和VDR基因的表达,两者的变化具有相关性(r=0.922,p<0.01)。
结论:
Ber具有明显调血脂作用,其机制可能与其上调VDR的表达,进而上调Insig-2基因表达,从而抑制肝脏中胆固醇的合成代谢有关。
Objective:
To investigate the effects of berberine (Ber) on cholesterol metabolism and its mechanism。
Methods:
1. SD rats were randomly divided into 5 groups: normal diet + pretreatment of solvent (ND); high-fat and high-cholesterol diet + pretreatment of solvent (HFD); high-fat and high-cholesterol diet + pretreatments of either lovastatin(LD) orBer low dose (BLD) and Ber high dose (BHD). The levels of total cholesterol(TC),low density lipoprotein cholesterol (LDL-C) , and high density lipoprotein cholesterol(HDL-C) in blood were detected eighteen weeks after rats were fed with high-fat and high-cholesterol diet and and pretreated with either solvent or drugs. The mRNA and protein expressions of Insulin-induced gene 2(Insig-2) in rat liver were quantified by RT-PCR and Western blot。
2. HepG2 cells were incubated with Ber at different concentrations (1, 3, 10, 30, 100μmol/L), or incubated with 3μmol/L Ber for various times (0, 12, 24, 48, 72 h). The gene expressions of Insig-2 and VDR from HepG2 cells treated by Ber were quantified by RT-PCR and Western blot.
Results:
1. The blood levels of TC and LDL-C were significantly lower in Ber groups than those in HFD, except HDL-C level that was shown to clearly increase after rats were pretreatment with Ber.
2.Low dose Ber pretreatment significantly increased Insig-2 gene mRNA and protein expressions from rat liver but high dose Ber pretreatment had no significantly effects on Insig-2 gene expression.
3.24 h after exposure to gradient concentrations of Ber, the gene expressions of Insig-2 and VDR in HepG2 cells showed a dualistic changes:the maximum values occurred at 3μmol/L, then the values declined gradually. The pretreatment with 3μmol/L Ber for different time made the gene expressions of Insig-2 and VDR significantly increased with the peak value at 48 h;the relative coefficient between them was 0.922 (p<0.01).
Conclusion:
Ber can significantly decrease the levels of hyperlidemia, the mechanism of which may be related to elevating expressions of Insig-2 and VDR genes in liver. As a result the cholesterol synthesis of liver was inhibited.
引文
[1]Kong WJ,Wei J, Alidi P,et al. Berberine is a novel cholesterol-lowering drug working through a unique mechanism distinct from statins[J]. Nat Med,2004, 10:(12):1344-1351.
[2]周丽斌,陈名道 等。小檗碱对脂肪细胞分化的影响[J]。中华医学杂志,2003,83(4): 338-340.
[3]Yabe D, Brown MS, Goldstein JL. Insig-2, a second endoplasmic reticulum protein that binds SCAP and blocks export of sterol regulatory element-binding proteins[J]. Proc Natl Acad Sci USA, 2002, 99(20): 12753-12758.
[4]Makishima M. Nuclear receptors as targets for drug development: regulation of cholesterol and bile acid metabolism by nuclear receptors[J]. J Pharmacol Sci.,2005, 97(2):177-183.
[5]Lee S, Lee DK, ChoI E, et al.. Identification of a functional vitamIn D response element in the murine Insig-2 promoter and Its potential role in the differentiation of 3T3-L1 Preadipocytes[J]. Mol Endocrinol.2005,19 (2): 399-408.
[6]Sever N, Song BL,Yabe D, et al. Insig-dependent ubiquitination and degradation of mammalian 3-hydroxy-3-methylglutaryl-CoA reductase stimulated by sterols and geranylgeraniol[J]. J Biol Chem,.2003, 278(52):52479-52490.
[7]Abidi P, Zhou Y, Jiang JD, et al.Extracellular signal-regulated kinase-dependent stabilization of hepatic low-density lipoprotein receptor mRNA by herbal medicine berberine[J]. Arterioscler Thromb Vasc Biol. 2005,25(10):2170-2176.
[8]Hubbert ML,Zhang Y, Lee FY,et al.Regulation of hepatic Insig-2 by the farnesoid X receptor[J].Mol Endocrinol.2007,21(6):1359-1369.
[9]Krapivner S,Popov S,Chernogubova E,et al.Insulin-induced gene 2(INSIG2) involvement in human adipocyte metabolism and body weight regulation[J].J Clin Endocrinol Metab.2008,[Epub ahead of print].
[10]Xie YH,Mo ZH,Chen K,et al.Effect of different glucose concentrations on the expressions of insig-1 and insig-2 mRNA during the differentiation of 3T3-L(1)acells[J].Zhong Nan Da Xue Xue Bao Yi Xue Ban.2008,33(3):238-244.
[11]Doggrell SA.Berberine—a novel approach to cholesterol lowering[J].Expert Opin Investig Drugs.2005,14(5):683-685.
[12]Jia X,Chen Y,Zidichouski J,et al.Co-administration of berberine and plant stanolssynergistically reduces plasama cholesterol in rats[J].Atherosclerosis.2008,[Epub ahead of print].
[13]Brusq JM,Ancellin N,Grondin P,et al.Inhibition of lipid sythesis through activation of AMP kinase:an additional mechanism for the hypolipidemic effects of berberine[J].J Lipid Res.2006,47(6):1281-1288.
[1]Blumberg JM, Tzameli I, Astapova I, et al. Complex role of the vitamin D receptor and its ligand in adipogenesis in 3T3-L1 cells [J]. J Biol Chem, 2006,281(16):11205-11213.
[2]林善锬,当代肾脏病学[M]。上海科技出版社,上海,2001, pp1048-1065.
[3]Ylikomi T,Laaksi I,Rulou Y,et al. Antipoliferative action of vitaminD[J].Vitam Horm,2002,64:357-406.
[4]Haussler MR,Whitfield GK,Hausshr CA,et a1.The nuclear vitamin D receptor:biological and molecular regulatory properties revealed[J].J Bone Miner Res ,1998,l3(3):325-349.
[5]Capiati DA,Tellez-Inon MT , Boland RL. Participation of protein kinase C alpha in 1 , 25-dihydroxy-vitaminD3 regulation of chick myoblast proliferation and differentiation[J].Mol Cell Endocrinol,1999,153(1-2):39- 45.
[5]Rostand SG.Ultraviolet light may contribute to geographic and racial blood pressure differences[J].Hypertension,1997,30(2pt1):l50-156.
[7]Zittermarm A .Vitamin D in preventive medicine:are we ignoring the evidence[J].Br J Nutr,2003,89(5):552-572.
[8]吴瑞萍,胡亚美,江载芳。诸福棠实用儿科学[M].。北京:人民卫生出版社,1997,557-559
[9]Parish JL . Topical vitamin D3 analogues : unappmved uses , dosages , or indications[J].Clin Dermatol,2002,20(5):558-562.
[10]Ailhaud G,Grimaldi P,Negnel R.Cellular and molecular aspects of adipose tissue development[J].Annu Rev Nutr,1992,12:207-233.
[11]Pilgrim C . DNA synthesis and differentiation in developing white adipose tissue[J].Dev Biol,1971,26(1):69- 76.
[12]Ono M ,Aratani Y,Kitagawa I,et a1.Ascorbic acid phosphatestimulates type IV collagen synthesis and accelerates adipose conversion of 3T3preadipocytes[J].Exp Cell Res,1987,87:309- 314.
[13]Miller WH Jr,Faust IM,Goldberger AC,et a1.Effects of severe long-term food deprivation and refeeding on adipose tissuecells in the rat[J].Am JPhysiol,1983,245(1):E74-E80.
[14]MacDougald OA,Lane MD.Transcriptional regulation of gene expression during adipocyte differentiation[J].Annu Rev Biochem,l995,64:345-373.
[15]Ntambi JM,Young-Cheul K.Adipocyte differentiation and gene expression[J].J Nutr,2000,130(12):3l22S-3126S.
[16]Horton JD,Shimomura I,Ikemoto S, et al. Overexpression of sterol regulatory element-binding protein-1a in mouse adipose tissue produces adipocyte hypertrophy, increased fatty acid secretion, and fatty liver[J].J Biol Chem, 2003, 278(38): 36652-36660.
[17]Ailhaud G,Grimaldi P,Negnel R.Cellular and molecular aspects of adipose tissue development[J].Annu Rev Nutr,1992,12:207-233.
[18]Sato M, Hiragun A.Demonstration of 1 alpha,25-dihydroxyvitamin D3 receptor-like molecule in ST 13 and 3T3 L1 preadipocytes and its inhibitory effects on preadipocyte differentiation[J]. J Cell Physiol, 1988, 135(3):545-550.
[19]Hida Y,Kawada T,Kayahashi S ,et al.Counteraction of retinoic acid and 1,25-dihydroxyvitamin D3 on up-regulation of adipocyte differentiation with PPARgamma ligand, an antidiabetic thiazolidinedione, in 3T3-L1 cells[J]. Iife Sci,1998,62(14):205-211.
[20]Ishida Y, Taniguchi H, Baba S.Possible involvement of 1 alpha, 25-dihydroxyvitamin D3 in proliferation and differentiation of 3T3-L1 cell[J].Biochem Biophys Res Commun,1988,151(3):1122-1127.
[21]Duque G, Macoritto M, Kremer R.1, 25-(OH)2D3 inhibits bone marrow adipogenesis in senescence accelerated mice (SAM-P/6) by decreasing the expression of peroxisome proliferator-activated receptor gamma2(PPARgamma2)[J].Exp Gerontol, 2004, 39(3):333-338.
[22]Katherine A, Kelly, Jeffrey M, et al.1,25-Dihydroxy Vitamin D3 inhibits adipocyte differentiation and gene expression in murine bone marrow stromal cell clones and primary cultures[J].Endocrinology,1998,139(5):52622-2628.
[23]Cianferotti L, Demay MB.VDR-mediated inhibition of DKK1 and SFRP2 suppresses adipogenic differentiation of murine bone marrow stromal cells[J].J Cell Biochem, 2007,101(1):80-88.
[24]Lee S,Lee DK,Choi E et al. Identification of a functional vitamin D responseelement in the murine Insig-2 promoter and its potential role in the differentiation of 3T3-L1 preadipocytes[J]. Mol Endocrinol, 2005,19(2):399-408.
[25]Kong J, Li YC.Molecular mechanism of 1,25-dihydroxyvitamin D3 inhibition of adipogenesis in 3T3-L1 cell[J].Am J Physiol Endocrinol Metab, 2006,290(5):E916-924.
[26]Bellows CG, Wang YH, Heersche JN et al.1, 25-dihydroxyvitamin D3 stimulates adipocyte differentiation in cultures of fetal rat calvaria cells: comparison with the effects of dexamethasone[J]. Endocrinology, 1994,134(5):2221-2229.
[27]Vu D, Ong JM,Clemens TL,et al. 1,25-Dihydroxyvitamin D induces lipoprotein lipase expression in 3T3-L1 cells in association with adipocyte differentiation [J]. Endocrinology , 1996, 137 (5) :1540-1544.
[2]周丽斌,陈名道 等。小檗碱对脂肪细胞分化的影响[J]。中华医学杂志,2003,83(4): 338-340.
[3]Yabe D, Brown MS, Goldstein JL. Insig-2, a second endoplasmic reticulum protein that binds SCAP and blocks export of sterol regulatory element-binding proteins[J]. Proc Natl Acad Sci USA, 2002, 99(20): 12753-12758.
[4]Makishima M. Nuclear receptors as targets for drug development: regulation of cholesterol and bile acid metabolism by nuclear receptors[J]. J Pharmacol Sci.,2005, 97(2):177-183.
[5]Lee S, Lee DK, ChoI E, et al.. Identification of a functional vitamIn D response element in the murine Insig-2 promoter and Its potential role in the differentiation of 3T3-L1 Preadipocytes[J]. Mol Endocrinol.2005,19 (2): 399-408.
[6]Sever N, Song BL,Yabe D, et al. Insig-dependent ubiquitination and degradation of mammalian 3-hydroxy-3-methylglutaryl-CoA reductase stimulated by sterols and geranylgeraniol[J]. J Biol Chem,.2003, 278(52):52479-52490.
[7]Abidi P, Zhou Y, Jiang JD, et al.Extracellular signal-regulated kinase-dependent stabilization of hepatic low-density lipoprotein receptor mRNA by herbal medicine berberine[J]. Arterioscler Thromb Vasc Biol. 2005,25(10):2170-2176.
[8]Hubbert ML,Zhang Y, Lee FY,et al.Regulation of hepatic Insig-2 by the farnesoid X receptor[J].Mol Endocrinol.2007,21(6):1359-1369.
[9]Krapivner S,Popov S,Chernogubova E,et al.Insulin-induced gene 2(INSIG2) involvement in human adipocyte metabolism and body weight regulation[J].J Clin Endocrinol Metab.2008,[Epub ahead of print].
[10]Xie YH,Mo ZH,Chen K,et al.Effect of different glucose concentrations on the expressions of insig-1 and insig-2 mRNA during the differentiation of 3T3-L(1)acells[J].Zhong Nan Da Xue Xue Bao Yi Xue Ban.2008,33(3):238-244.
[11]Doggrell SA.Berberine—a novel approach to cholesterol lowering[J].Expert Opin Investig Drugs.2005,14(5):683-685.
[12]Jia X,Chen Y,Zidichouski J,et al.Co-administration of berberine and plant stanolssynergistically reduces plasama cholesterol in rats[J].Atherosclerosis.2008,[Epub ahead of print].
[13]Brusq JM,Ancellin N,Grondin P,et al.Inhibition of lipid sythesis through activation of AMP kinase:an additional mechanism for the hypolipidemic effects of berberine[J].J Lipid Res.2006,47(6):1281-1288.
[1]Blumberg JM, Tzameli I, Astapova I, et al. Complex role of the vitamin D receptor and its ligand in adipogenesis in 3T3-L1 cells [J]. J Biol Chem, 2006,281(16):11205-11213.
[2]林善锬,当代肾脏病学[M]。上海科技出版社,上海,2001, pp1048-1065.
[3]Ylikomi T,Laaksi I,Rulou Y,et al. Antipoliferative action of vitaminD[J].Vitam Horm,2002,64:357-406.
[4]Haussler MR,Whitfield GK,Hausshr CA,et a1.The nuclear vitamin D receptor:biological and molecular regulatory properties revealed[J].J Bone Miner Res ,1998,l3(3):325-349.
[5]Capiati DA,Tellez-Inon MT , Boland RL. Participation of protein kinase C alpha in 1 , 25-dihydroxy-vitaminD3 regulation of chick myoblast proliferation and differentiation[J].Mol Cell Endocrinol,1999,153(1-2):39- 45.
[5]Rostand SG.Ultraviolet light may contribute to geographic and racial blood pressure differences[J].Hypertension,1997,30(2pt1):l50-156.
[7]Zittermarm A .Vitamin D in preventive medicine:are we ignoring the evidence[J].Br J Nutr,2003,89(5):552-572.
[8]吴瑞萍,胡亚美,江载芳。诸福棠实用儿科学[M].。北京:人民卫生出版社,1997,557-559
[9]Parish JL . Topical vitamin D3 analogues : unappmved uses , dosages , or indications[J].Clin Dermatol,2002,20(5):558-562.
[10]Ailhaud G,Grimaldi P,Negnel R.Cellular and molecular aspects of adipose tissue development[J].Annu Rev Nutr,1992,12:207-233.
[11]Pilgrim C . DNA synthesis and differentiation in developing white adipose tissue[J].Dev Biol,1971,26(1):69- 76.
[12]Ono M ,Aratani Y,Kitagawa I,et a1.Ascorbic acid phosphatestimulates type IV collagen synthesis and accelerates adipose conversion of 3T3preadipocytes[J].Exp Cell Res,1987,87:309- 314.
[13]Miller WH Jr,Faust IM,Goldberger AC,et a1.Effects of severe long-term food deprivation and refeeding on adipose tissuecells in the rat[J].Am JPhysiol,1983,245(1):E74-E80.
[14]MacDougald OA,Lane MD.Transcriptional regulation of gene expression during adipocyte differentiation[J].Annu Rev Biochem,l995,64:345-373.
[15]Ntambi JM,Young-Cheul K.Adipocyte differentiation and gene expression[J].J Nutr,2000,130(12):3l22S-3126S.
[16]Horton JD,Shimomura I,Ikemoto S, et al. Overexpression of sterol regulatory element-binding protein-1a in mouse adipose tissue produces adipocyte hypertrophy, increased fatty acid secretion, and fatty liver[J].J Biol Chem, 2003, 278(38): 36652-36660.
[17]Ailhaud G,Grimaldi P,Negnel R.Cellular and molecular aspects of adipose tissue development[J].Annu Rev Nutr,1992,12:207-233.
[18]Sato M, Hiragun A.Demonstration of 1 alpha,25-dihydroxyvitamin D3 receptor-like molecule in ST 13 and 3T3 L1 preadipocytes and its inhibitory effects on preadipocyte differentiation[J]. J Cell Physiol, 1988, 135(3):545-550.
[19]Hida Y,Kawada T,Kayahashi S ,et al.Counteraction of retinoic acid and 1,25-dihydroxyvitamin D3 on up-regulation of adipocyte differentiation with PPARgamma ligand, an antidiabetic thiazolidinedione, in 3T3-L1 cells[J]. Iife Sci,1998,62(14):205-211.
[20]Ishida Y, Taniguchi H, Baba S.Possible involvement of 1 alpha, 25-dihydroxyvitamin D3 in proliferation and differentiation of 3T3-L1 cell[J].Biochem Biophys Res Commun,1988,151(3):1122-1127.
[21]Duque G, Macoritto M, Kremer R.1, 25-(OH)2D3 inhibits bone marrow adipogenesis in senescence accelerated mice (SAM-P/6) by decreasing the expression of peroxisome proliferator-activated receptor gamma2(PPARgamma2)[J].Exp Gerontol, 2004, 39(3):333-338.
[22]Katherine A, Kelly, Jeffrey M, et al.1,25-Dihydroxy Vitamin D3 inhibits adipocyte differentiation and gene expression in murine bone marrow stromal cell clones and primary cultures[J].Endocrinology,1998,139(5):52622-2628.
[23]Cianferotti L, Demay MB.VDR-mediated inhibition of DKK1 and SFRP2 suppresses adipogenic differentiation of murine bone marrow stromal cells[J].J Cell Biochem, 2007,101(1):80-88.
[24]Lee S,Lee DK,Choi E et al. Identification of a functional vitamin D responseelement in the murine Insig-2 promoter and its potential role in the differentiation of 3T3-L1 preadipocytes[J]. Mol Endocrinol, 2005,19(2):399-408.
[25]Kong J, Li YC.Molecular mechanism of 1,25-dihydroxyvitamin D3 inhibition of adipogenesis in 3T3-L1 cell[J].Am J Physiol Endocrinol Metab, 2006,290(5):E916-924.
[26]Bellows CG, Wang YH, Heersche JN et al.1, 25-dihydroxyvitamin D3 stimulates adipocyte differentiation in cultures of fetal rat calvaria cells: comparison with the effects of dexamethasone[J]. Endocrinology, 1994,134(5):2221-2229.
[27]Vu D, Ong JM,Clemens TL,et al. 1,25-Dihydroxyvitamin D induces lipoprotein lipase expression in 3T3-L1 cells in association with adipocyte differentiation [J]. Endocrinology , 1996, 137 (5) :1540-1544.