雷公藤多糖诱导胰腺干细胞成胰岛样细胞团（英文）
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摘要
背景:胰岛细胞来源不足说明胰岛细胞移植治疗糖尿病不能满足临床需要。因此,体外诱导胰腺干细胞分化为胰岛细胞成为研究的热点。目的:观察雷公藤多糖对小鼠胰岛来源的胰腺干细胞的诱导分化作用,从而探索中药诱导胰腺干细胞分化为胰岛β细胞的作用。方法:采用雷公藤多糖体外诱导经纯化的小鼠胰腺干细胞分化为胰岛细胞。胰岛样细胞团进行形态学观察,双硫腙染色和Western blot分析。结果与结论:①细胞形态学、细胞生长特性及免疫细胞化学染色显示,实验不仅得到了纯度高的小鼠胰腺干细胞,而且该细胞经雷公藤多糖诱导后能形成球形的胰岛样结构,有细长的蒂与培养瓶底部相连,双硫腙染色呈铁红色;②Western-blot检测显示,胰岛样细胞团中有β细胞素蛋白的表达;③结果表明,小鼠胰腺干细胞可以体外经雷公藤多糖诱导分化为含有β细胞的胰岛样细胞团。
BACKGROUND: Inadequate sources of islet cells mean that islet cell transplantation for diabetes cannot meet the clinical demand. Therefore, in vitro induction of pancreatic stem cells to differentiate into islets has become a focus of research. OBJECTIVE: To study the effect of Tripterygium wilfordii polysaccharides on the differentiation of pancreatic stem cells from islets in mice, so as to explore the effect of traditional Chinese medicine on the differentiation of pancreatic stem cells into pancreatic beta cells. METHODS: Tripterygium wilfordii polysaccharide was used to induce the differentiation of purified mouse pancreatic stem cells into islets in vitro. The islet-like cell clusters then underwent morphologic observation, dithizone(DTZ) staining, and western blot analysis. RESULTS AND CONCLUSION: Cell morphology, cell growth characteristics and immunocytochemical staining showed that mouse pancreatic stem cells were obtained. They were induced by Tripterygium wilfordii polysaccharide into spherical islet-like structures, which had a spindly pedicle connected with the bottom of the culture flask, and were DTZ-stained to iron red. Western blot assay detected β-cytokine proteins in the islet-like cell clusters. These findings confirm that mouse pancreatic stem cells can be induced to differentiate into islet-like cell clusters containing β cells in vitro by Tripterygium wilfordii polysaccharide.
BACKGROUND: Inadequate sources of islet cells mean that islet cell transplantation for diabetes cannot meet the clinical demand. Therefore, in vitro induction of pancreatic stem cells to differentiate into islets has become a focus of research. OBJECTIVE: To study the effect of Tripterygium wilfordii polysaccharides on the differentiation of pancreatic stem cells from islets in mice, so as to explore the effect of traditional Chinese medicine on the differentiation of pancreatic stem cells into pancreatic beta cells. METHODS: Tripterygium wilfordii polysaccharide was used to induce the differentiation of purified mouse pancreatic stem cells into islets in vitro. The islet-like cell clusters then underwent morphologic observation, dithizone(DTZ) staining, and western blot analysis. RESULTS AND CONCLUSION: Cell morphology, cell growth characteristics and immunocytochemical staining showed that mouse pancreatic stem cells were obtained. They were induced by Tripterygium wilfordii polysaccharide into spherical islet-like structures, which had a spindly pedicle connected with the bottom of the culture flask, and were DTZ-stained to iron red. Western blot assay detected β-cytokine proteins in the islet-like cell clusters. These findings confirm that mouse pancreatic stem cells can be induced to differentiate into islet-like cell clusters containing β cells in vitro by Tripterygium wilfordii polysaccharide.
引文
[1]Hernandez-Perez M,Chopra G.Inhibition of12/15-lipoxygenase protects againstβ-cell oxidative stress and glycemic deterioration in mouse models of type 1diabetes.Diabetes.2017;66(11):2875-2887.
[2]Gao YR.Internal Medicine.5th ed.Beijing:People’s Medical Publishing House,2002.
[3]Rosenberg L,Vinik AI,Pittenger GL,Rafaeloff R,Duguid WP.Islet cell regeneration in the diabetic hamster pancreas with restoration of normoglycaemia can be induced by a local growth factor(s).Diabetologia.1996;39:256-262.
[4]Schmied BM,Liu G,Matsuzaki H,Ulrich A,et al.Differentiation of islet cells in long-term culture.Pancreas.2000;20:337-347.
[5]Liu X,Shan W,Zeng RX,et al.Differentiation of rat bone marrow mesenchymal stem cells into neuron-like cells in Lycium barbarum polysaccharide.Zhongguo Zuzhi Gongcheng Yanjiu.2009;13:2667-2672.
[6]Liu JK,Song YF,Liu ZY,et al.Cordyceps polysaccharide induces differentiation of adult rat mesenchymal stem cells into a hepatocyte lineage in vitro.Zhongguo Zuzhi Gongcheng Yanjiu.2012;16:9221-9225.
[7]The Ministry of Science and Technology of the People’s Republic of China.Guidance Suggestions for the Care and Use of Laboratory Animals.2006-09-30.
[8]Song ZS,Gu KJ.Morphological changes of adult human pancreatic stem cells transdifferentiating into islet cells.Waike Lilun yu Shijian.2002;7:196-200.
[9]Rao ZX,Qin ML,Zhou DS.Islet stem cells in vitro culture and growth characteristics.Prog Anat Sci.2003;9:16-48.
[10]Shiroi A,Yoshikawa M,Yokota H,et al.Identification of insulin-producing cells derived from embryonic stem cells by zinc-chelating dithizone.Stem Cells.2002;20:284-292.
[11]Zhang XL,Zhang SY,Sun Y,et al.Serum-free culture and stability during subculture of Vero cells.Zhongguo Shengwuzhipinxue Zazhi.2017;30(1):68-73.
[12]Kou YL,Hu LX,Li Q,et al.Method for culturing and amplifying pancreatic stem cells in vitro.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu.2011;15(10):819-1822.
[13]Cen YH,He GZ,Huang B,et al.Isolation,culture and natural differentiation of pancreatic stem cells from newborn Kunming mice.Zhongguo Zuzhi Gongcheng Yanjiu.2012;16:1019-1023.
[14]Mc Kinnon CM,Docherty K.Pancreatic duodenal homeobox-1,pdx-1,a major regulator of beta cell identity and function.Diabetologia.2001;44:1203-1214.
[15]Jonsson J,Carlsson L,Edlund T,et al.Insulin-promoter-factor 1 is required for pancreas development in mice.Nature.1994;371:606-609.
[16]Offield MF,Jetton TL,Labosky PA,et al.PDX-1 is required for pancreatic outgrowth and differentiation of the rostral duodenum.Development.1996;122:983-995.
[17]Zhang XM.Pharmacological action and clinical application research progress of tripterygium wilfordii.Linchuang Heli Yongyao.2012;5:180.
[18]Zhang LH,Ouyang KA,Zhen YL.The experimental observation of Tripterygium wilfordii hypoglycemic in mice.Fujian Med J.1990;12:39-40.
[19]Ouyang LL,Zhou ZG,Peng J,et al.Preliminary clinical study on tripterygium intervention in LADA patients.2000;8(1):7-9.
[20]Hu J,Feng M,Yang H,et al.Hematopoiesis reconstruction in mice of hematopoietic aplasia by transplanting hematopoietic stem/progenitor cells mobilized by Angelic polysaccharides.Disan Junyidaxue Xuebao.2007;29:2236-2239.
[21]Wu H,Zhao J,Li FF,et al.Study on hematopoietic reconstruction for bone marrow depression mice with the transplantation of mobilized peripheral blood stem/progenitor cells by ginseng polysaccharide.Chongqing Yikedaxue Xuebao.2006;31:804-807.
[22]Cai GX,Lin L,Liu BY,et al.Effect of Rehmannia glutinosa polysaccharides on the differentiation of bone marrow mesenchymal stem cells into nerve cells.Zhongguo Linchuang Kangfu.2005;9:46-47.
[23]Zhang YQ.Effect of Rehmannia glutinosa polysaccharides(RPS)on the differentiation of adipose tissue-derived mesenchymal stem cells(ADMSCs)into cardiomyocytes.Beijing:Chinese PLA Postgraduate Medical School.2008.
[24]Miyagawa J,Hanafusa O,Sasada R,et al.Immunohistochemical localization of betacellulin,a new member of the EGF family,in normal human pancreas and islet tumor cells.Endocr J.1999;46:755-764.
[25]Mashima H,Ohnishi H,Wakabayashi K,et al.Betacellulin and activin A coordinately convert amylase-secreting AR42J cells into insulin-secreting cells.J Clin Invest 1996;97:1647-1654.
[26]Demeterco C,Beattie GM,Dib SA,et al.A role for activin A and betacellulin in human fetal pancreatic cell differentiation and growth.J Clin Endocrinol Metab.2000;85:3892-3897.
[2]Gao YR.Internal Medicine.5th ed.Beijing:People’s Medical Publishing House,2002.
[3]Rosenberg L,Vinik AI,Pittenger GL,Rafaeloff R,Duguid WP.Islet cell regeneration in the diabetic hamster pancreas with restoration of normoglycaemia can be induced by a local growth factor(s).Diabetologia.1996;39:256-262.
[4]Schmied BM,Liu G,Matsuzaki H,Ulrich A,et al.Differentiation of islet cells in long-term culture.Pancreas.2000;20:337-347.
[5]Liu X,Shan W,Zeng RX,et al.Differentiation of rat bone marrow mesenchymal stem cells into neuron-like cells in Lycium barbarum polysaccharide.Zhongguo Zuzhi Gongcheng Yanjiu.2009;13:2667-2672.
[6]Liu JK,Song YF,Liu ZY,et al.Cordyceps polysaccharide induces differentiation of adult rat mesenchymal stem cells into a hepatocyte lineage in vitro.Zhongguo Zuzhi Gongcheng Yanjiu.2012;16:9221-9225.
[7]The Ministry of Science and Technology of the People’s Republic of China.Guidance Suggestions for the Care and Use of Laboratory Animals.2006-09-30.
[8]Song ZS,Gu KJ.Morphological changes of adult human pancreatic stem cells transdifferentiating into islet cells.Waike Lilun yu Shijian.2002;7:196-200.
[9]Rao ZX,Qin ML,Zhou DS.Islet stem cells in vitro culture and growth characteristics.Prog Anat Sci.2003;9:16-48.
[10]Shiroi A,Yoshikawa M,Yokota H,et al.Identification of insulin-producing cells derived from embryonic stem cells by zinc-chelating dithizone.Stem Cells.2002;20:284-292.
[11]Zhang XL,Zhang SY,Sun Y,et al.Serum-free culture and stability during subculture of Vero cells.Zhongguo Shengwuzhipinxue Zazhi.2017;30(1):68-73.
[12]Kou YL,Hu LX,Li Q,et al.Method for culturing and amplifying pancreatic stem cells in vitro.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu.2011;15(10):819-1822.
[13]Cen YH,He GZ,Huang B,et al.Isolation,culture and natural differentiation of pancreatic stem cells from newborn Kunming mice.Zhongguo Zuzhi Gongcheng Yanjiu.2012;16:1019-1023.
[14]Mc Kinnon CM,Docherty K.Pancreatic duodenal homeobox-1,pdx-1,a major regulator of beta cell identity and function.Diabetologia.2001;44:1203-1214.
[15]Jonsson J,Carlsson L,Edlund T,et al.Insulin-promoter-factor 1 is required for pancreas development in mice.Nature.1994;371:606-609.
[16]Offield MF,Jetton TL,Labosky PA,et al.PDX-1 is required for pancreatic outgrowth and differentiation of the rostral duodenum.Development.1996;122:983-995.
[17]Zhang XM.Pharmacological action and clinical application research progress of tripterygium wilfordii.Linchuang Heli Yongyao.2012;5:180.
[18]Zhang LH,Ouyang KA,Zhen YL.The experimental observation of Tripterygium wilfordii hypoglycemic in mice.Fujian Med J.1990;12:39-40.
[19]Ouyang LL,Zhou ZG,Peng J,et al.Preliminary clinical study on tripterygium intervention in LADA patients.2000;8(1):7-9.
[20]Hu J,Feng M,Yang H,et al.Hematopoiesis reconstruction in mice of hematopoietic aplasia by transplanting hematopoietic stem/progenitor cells mobilized by Angelic polysaccharides.Disan Junyidaxue Xuebao.2007;29:2236-2239.
[21]Wu H,Zhao J,Li FF,et al.Study on hematopoietic reconstruction for bone marrow depression mice with the transplantation of mobilized peripheral blood stem/progenitor cells by ginseng polysaccharide.Chongqing Yikedaxue Xuebao.2006;31:804-807.
[22]Cai GX,Lin L,Liu BY,et al.Effect of Rehmannia glutinosa polysaccharides on the differentiation of bone marrow mesenchymal stem cells into nerve cells.Zhongguo Linchuang Kangfu.2005;9:46-47.
[23]Zhang YQ.Effect of Rehmannia glutinosa polysaccharides(RPS)on the differentiation of adipose tissue-derived mesenchymal stem cells(ADMSCs)into cardiomyocytes.Beijing:Chinese PLA Postgraduate Medical School.2008.
[24]Miyagawa J,Hanafusa O,Sasada R,et al.Immunohistochemical localization of betacellulin,a new member of the EGF family,in normal human pancreas and islet tumor cells.Endocr J.1999;46:755-764.
[25]Mashima H,Ohnishi H,Wakabayashi K,et al.Betacellulin and activin A coordinately convert amylase-secreting AR42J cells into insulin-secreting cells.J Clin Invest 1996;97:1647-1654.
[26]Demeterco C,Beattie GM,Dib SA,et al.A role for activin A and betacellulin in human fetal pancreatic cell differentiation and growth.J Clin Endocrinol Metab.2000;85:3892-3897.