人脂肪间充质干细胞尾静脉移植治疗大鼠急性肝衰竭
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摘要
背景:间充质干细胞对肝衰竭大鼠的肝脏具有保护及修复作用,但具体机制尚未完全明确。目的:观察人脂肪间充质干细胞尾静脉注射对急性肝衰竭大鼠肝脏损伤的保护作用,并探讨其可能的机制。方法:将36只SD大鼠(购自青岛市大任富成畜牧有限公司)随机等分为对照组、模型组和移植组。模型组和移植组大鼠通过腹腔注射D-氨基半乳糖建立急性肝衰竭模型。造模1 d后,移植组注射人脂肪间充质干细胞悬液,模型组注射等量生理盐水。细胞移植1 d和3 d,检测血清中丙氨酸氨基转移酶、天冬氨酸氨基转移酶、总胆红素水平。细胞移植3 d,检测血清中肿瘤坏死因子α、白细胞介素6、白细胞介素10水平;苏木精-伊红染色观察肝脏病理形态学变化;Western blot检测肝组织中糖原合成酶激酶3β蛋白的表达。结果与结论:(1)与模型组相比,移植组血清中丙氨酸氨基转移酶、天冬氨酸氨基转移酶、总胆红素、肿瘤坏死因子α、白细胞介素6和白细胞介素10水平降低(P<0.05);(2)移植组肝脏组织炎症反应和变性坏死程度较模型组减轻;(3)移植组肝组织中糖原合成酶激酶3β活性较模型组减弱(P <0.05);(4)结果表明,人脂肪间充质干细胞可以减轻急性肝衰竭大鼠肝脏炎症反应和病理损伤,改善肝脏功能,其机制可能与抑制糖原合成酶激酶3β活性有关。
BACKGROUND: Mesenchymal stem cells can protect and repair the liver of rats with liver failure, but the mechanisms are not completely clear. OBJECTIVE: To explore the protective effects and related mechanisms of intravenous injection of human adipose-derived mesenchymal stem cells on acute liver failure in rats. METHODS: Thirty-six Sprague-Dawley rats(provided by Qingdao Daren Fucheng Animal Husbandry Co., Ltd. in China) were randomly divided into control group, model group and transplantation group. Animal models of acute liver failure were established by intraperitoneal injection of D-galactosamine in the model group and the transplantation group. One day after modeling, the rats in the transplantation group were injected with human adipose-derived mesenchymal stem cell suspension, and those in the model group were injected with the same amount of saline. After 1 and 3 days of cell transplantation, the serum levels of alanine aminotransferase, aspartate aminotransferase, and total bilirubin were measured. Three days after cell transplantation, the serum levels of tumor necrosis factor-α, interleukin-6 and interleukin-10 were detected, the pathological changes of the rat liver were observed by hematoxylin-eosin staining, and the activity of glycogen synthase kinase-3β protein in the liver tissue was detected by western blot. RESULTS AND CONCLUSION: Compared with the model group, there was a significant reduction in the serum levels of alanine aminotransferase, aspartate aminotransferase, total bilirubin, tumor necrosis factor-α, interleukin-6 and interleukin-10 in the transplantation group(P < 0.05). Inflammation and necrosis of liver tissues in the transplant group were alleviated compared with the model group. The activity of glycogen synthase kinase 3β in the liver tissue of the transplanted group was lower than that of the model group(P < 0.05). Overall, these results indicate that human adipose-derived mesenchymal stem cells can alleviate hepatic inflammation and pathological injury, and improve the liver function in rats with acute hepatic failure. Moreover, the mechanism may be related to the inhibition of glycogen synthase kinase 3β activity.
BACKGROUND: Mesenchymal stem cells can protect and repair the liver of rats with liver failure, but the mechanisms are not completely clear. OBJECTIVE: To explore the protective effects and related mechanisms of intravenous injection of human adipose-derived mesenchymal stem cells on acute liver failure in rats. METHODS: Thirty-six Sprague-Dawley rats(provided by Qingdao Daren Fucheng Animal Husbandry Co., Ltd. in China) were randomly divided into control group, model group and transplantation group. Animal models of acute liver failure were established by intraperitoneal injection of D-galactosamine in the model group and the transplantation group. One day after modeling, the rats in the transplantation group were injected with human adipose-derived mesenchymal stem cell suspension, and those in the model group were injected with the same amount of saline. After 1 and 3 days of cell transplantation, the serum levels of alanine aminotransferase, aspartate aminotransferase, and total bilirubin were measured. Three days after cell transplantation, the serum levels of tumor necrosis factor-α, interleukin-6 and interleukin-10 were detected, the pathological changes of the rat liver were observed by hematoxylin-eosin staining, and the activity of glycogen synthase kinase-3β protein in the liver tissue was detected by western blot. RESULTS AND CONCLUSION: Compared with the model group, there was a significant reduction in the serum levels of alanine aminotransferase, aspartate aminotransferase, total bilirubin, tumor necrosis factor-α, interleukin-6 and interleukin-10 in the transplantation group(P < 0.05). Inflammation and necrosis of liver tissues in the transplant group were alleviated compared with the model group. The activity of glycogen synthase kinase 3β in the liver tissue of the transplanted group was lower than that of the model group(P < 0.05). Overall, these results indicate that human adipose-derived mesenchymal stem cells can alleviate hepatic inflammation and pathological injury, and improve the liver function in rats with acute hepatic failure. Moreover, the mechanism may be related to the inhibition of glycogen synthase kinase 3β activity.
引文
[1]李兰娟,段钟平.肝衰竭诊治指南(2012年版)[J].中华肝脏病杂志,2013,21(3):210-216.
[2]刘景丰,张翔.肝衰竭的肝移植治疗进展[J].中华肝脏病杂志,2017,25(9):655-658.
[3]Squillaro T,Peluso G,Galderisi U.Clinical Trials With Mesenchymal Stem Cells:An Update.Cell Transplant.2016;25(5):829-848.
[4]Fierabracci A,Del Fattore A,Luciano R,et al.Recent advances in mesenchymal stem cell immunomodulation:the role of microvesicles.Cell Transplant.2015;24(2):133-149.
[5]Wong SP,Rowley JE,Redpath AN,et al.Pericytes,mesenchymal stem cells and their contributions to tissue repair.Pharmacol Ther.2015;151:107-120.
[6]Volarevic V,Nurkovic J,Arsenijevic N,et al.Concise review:Therapeutic potential of mesenchymal stem cells for the treatment of acute liver failure and cirrhosis.Stem Cells.2014;32(11):2818-2823.
[7]Liu WH,Song FQ,Ren LN,et al.The multiple functional roles of mesenchymal stem cells in participating in treating liver diseases.J Cell Mol Med.2015;19(3):511-520.
[8]Lee CW,Chen YF,Wu HH,et al.Historical Perspectives and Advances in Mesenchymal Stem Cell Research for the Treatment of Liver Diseases.Gastroenterology.2018;154(1):46-56.
[9]Wu Z,Han M,Chen T,et al.Acute liver failure:mechanisms of immune-mediated liver injury.Liver Int.2010;30(6):782-794.
[10]Wang X,Ning Q.Immune mediated liver failure.EXCLI J.2014;13:1131-1144.
[11]李晓东.HBV相关慢加急性肝衰竭免疫病理发病机制研究进展[J].实用肝脏病杂志,2015,18(3):317-320.
[12]Bassi G,Pacelli L,Carusone R,et al.Adipose-derived stromal cells(ASCs).Transfus Apher Sci.2012;47(2):193-198.
[13]Dominici M,Le Blanc K,Mueller I,et al.Minimal criteria for defining multipotent mesenchymal stromal cells.The International Society for Cellular Therapy position statement.Cytotherapy.2006;8(4):315-317.
[14]Bourin P,Bunnell BA,Casteilla L,et al.Stromal cells from the adipose tissue-derived stromal vascular fraction and culture expanded adipose tissue-derived stromal/stem cells:a joint statement of the International Federation for Adipose Therapeutics and Science(IFATS)and the International Society for Cellular Therapy(ISCT).Cytotherapy.2013;15(6):641-648.
[15]Liu WH,Song FQ,Ren LN,et al.The multiple functional roles of mesenchymal stem cells in participating in treating liver diseases.J Cell Mol Med.2015;19(3):511-520.
[16]Bajek A,Gurtowska N,Olkowska J,et al.Adipose-Derived Stem Cells as a Tool in Cell-Based Therapies.Arch Immunol Ther Exp(Warsz).2016;64(6):443-454.
[17]Tabatabaei Qomi R,Sheykhhasan M.Adipose-derived stromal cell in regenerative medicine:A review.World J Stem Cells.2017;9(8):107-117.
[18]Chen G,Jin Y,Shi X,et al.Adipose-derived stem cell-based treatment for acute liver failure.Stem Cell Res Ther.2015;6:40.
[19]Deng L,Kong X,Liu G,et al.Transplantation of Adipose-Derived Mesenchymal Stem Cells Efficiently Rescues Thioacetamide-Induced Acute Liver Failure in Mice.Transplant Proc.2016;48(6):2208-2215.
[20]史光军,张亚东,胡音音,等.脂肪间充质干细胞移植上调肝脏增殖细胞核抗原表达促进肝细胞的再生[J].中国组织工程研究,2017,21(17):2690-2695.
[21]Grimes CA,Jope RS.The multifaceted roles of glycogen synthase kinase 3beta in cellular signaling.Prog Neurobiol.2001;65(4):391-426.
[22]Beurel E,Grieco SF,Jope RS.Glycogen synthase kinase-3(GSK3):regulation,actions,and diseases.Pharmacol Ther.2015;148:114-131.
[23]Mishra R,Nagini S,Rana A.Expression and inactivation of glycogen synthase kinase 3 alpha/beta and their association with the expression of cyclin D1 and p53 in oral squamous cell carcinoma progression.Mol Cancer.2015;14:20.
[24]Beurel E,Michalek SM,Jope RS.Innate and adaptive immune responses regulated by glycogen synthase kinase-3(GSK3).Trends Immunol.2010;31(1):24-31.
[25]谈志丽,钟欢,朱彤,等.糖原合成激酶3β在急性肝衰竭免疫炎症反应中的作用及机制研究进展[J].免疫学杂志,2017,33(6):547-552.
[26]Chen L,Ren F,Zhang H,et al.Inhibition of glycogen synthase kinase 3βameliorates D-GalN/LPS-induced liver injury by reducing endoplasmic reticulum stress-triggered apoptosis.PLoS One.2012;7(9):e45202.
[27]Zhang H,Wang W,Fang H,et al.GSK-3βinhibition attenuates CLP-induced liver injury by reducing inflammation and hepatic cell apoptosis.Mediators Inflamm.2014;2014:629507.
[28]张向颖,任锋,魏琳琳,等.糖原合成酶激酶3β在乙型重型肝炎肝衰竭中的作用研究[J].中华肝脏病杂志,2016,24(4):265-269.
[29]Ren F,Zhang L,Zhang X,et al.Inhibition of glycogen synthase kinase 3βpromotes autophagy to protect mice from acute liver failure mediated by peroxisome proliferatoractivated receptorα.Cell Death Dis.2016;7:e2151.
[30]杨丙章,任锋,温韬,等.糖原合成酶激酶-3β在D-GalN/LPS诱导的小鼠急性肝衰竭中的作用[J].世界华人消化杂志,2012,20(36):3656-3662.
[2]刘景丰,张翔.肝衰竭的肝移植治疗进展[J].中华肝脏病杂志,2017,25(9):655-658.
[3]Squillaro T,Peluso G,Galderisi U.Clinical Trials With Mesenchymal Stem Cells:An Update.Cell Transplant.2016;25(5):829-848.
[4]Fierabracci A,Del Fattore A,Luciano R,et al.Recent advances in mesenchymal stem cell immunomodulation:the role of microvesicles.Cell Transplant.2015;24(2):133-149.
[5]Wong SP,Rowley JE,Redpath AN,et al.Pericytes,mesenchymal stem cells and their contributions to tissue repair.Pharmacol Ther.2015;151:107-120.
[6]Volarevic V,Nurkovic J,Arsenijevic N,et al.Concise review:Therapeutic potential of mesenchymal stem cells for the treatment of acute liver failure and cirrhosis.Stem Cells.2014;32(11):2818-2823.
[7]Liu WH,Song FQ,Ren LN,et al.The multiple functional roles of mesenchymal stem cells in participating in treating liver diseases.J Cell Mol Med.2015;19(3):511-520.
[8]Lee CW,Chen YF,Wu HH,et al.Historical Perspectives and Advances in Mesenchymal Stem Cell Research for the Treatment of Liver Diseases.Gastroenterology.2018;154(1):46-56.
[9]Wu Z,Han M,Chen T,et al.Acute liver failure:mechanisms of immune-mediated liver injury.Liver Int.2010;30(6):782-794.
[10]Wang X,Ning Q.Immune mediated liver failure.EXCLI J.2014;13:1131-1144.
[11]李晓东.HBV相关慢加急性肝衰竭免疫病理发病机制研究进展[J].实用肝脏病杂志,2015,18(3):317-320.
[12]Bassi G,Pacelli L,Carusone R,et al.Adipose-derived stromal cells(ASCs).Transfus Apher Sci.2012;47(2):193-198.
[13]Dominici M,Le Blanc K,Mueller I,et al.Minimal criteria for defining multipotent mesenchymal stromal cells.The International Society for Cellular Therapy position statement.Cytotherapy.2006;8(4):315-317.
[14]Bourin P,Bunnell BA,Casteilla L,et al.Stromal cells from the adipose tissue-derived stromal vascular fraction and culture expanded adipose tissue-derived stromal/stem cells:a joint statement of the International Federation for Adipose Therapeutics and Science(IFATS)and the International Society for Cellular Therapy(ISCT).Cytotherapy.2013;15(6):641-648.
[15]Liu WH,Song FQ,Ren LN,et al.The multiple functional roles of mesenchymal stem cells in participating in treating liver diseases.J Cell Mol Med.2015;19(3):511-520.
[16]Bajek A,Gurtowska N,Olkowska J,et al.Adipose-Derived Stem Cells as a Tool in Cell-Based Therapies.Arch Immunol Ther Exp(Warsz).2016;64(6):443-454.
[17]Tabatabaei Qomi R,Sheykhhasan M.Adipose-derived stromal cell in regenerative medicine:A review.World J Stem Cells.2017;9(8):107-117.
[18]Chen G,Jin Y,Shi X,et al.Adipose-derived stem cell-based treatment for acute liver failure.Stem Cell Res Ther.2015;6:40.
[19]Deng L,Kong X,Liu G,et al.Transplantation of Adipose-Derived Mesenchymal Stem Cells Efficiently Rescues Thioacetamide-Induced Acute Liver Failure in Mice.Transplant Proc.2016;48(6):2208-2215.
[20]史光军,张亚东,胡音音,等.脂肪间充质干细胞移植上调肝脏增殖细胞核抗原表达促进肝细胞的再生[J].中国组织工程研究,2017,21(17):2690-2695.
[21]Grimes CA,Jope RS.The multifaceted roles of glycogen synthase kinase 3beta in cellular signaling.Prog Neurobiol.2001;65(4):391-426.
[22]Beurel E,Grieco SF,Jope RS.Glycogen synthase kinase-3(GSK3):regulation,actions,and diseases.Pharmacol Ther.2015;148:114-131.
[23]Mishra R,Nagini S,Rana A.Expression and inactivation of glycogen synthase kinase 3 alpha/beta and their association with the expression of cyclin D1 and p53 in oral squamous cell carcinoma progression.Mol Cancer.2015;14:20.
[24]Beurel E,Michalek SM,Jope RS.Innate and adaptive immune responses regulated by glycogen synthase kinase-3(GSK3).Trends Immunol.2010;31(1):24-31.
[25]谈志丽,钟欢,朱彤,等.糖原合成激酶3β在急性肝衰竭免疫炎症反应中的作用及机制研究进展[J].免疫学杂志,2017,33(6):547-552.
[26]Chen L,Ren F,Zhang H,et al.Inhibition of glycogen synthase kinase 3βameliorates D-GalN/LPS-induced liver injury by reducing endoplasmic reticulum stress-triggered apoptosis.PLoS One.2012;7(9):e45202.
[27]Zhang H,Wang W,Fang H,et al.GSK-3βinhibition attenuates CLP-induced liver injury by reducing inflammation and hepatic cell apoptosis.Mediators Inflamm.2014;2014:629507.
[28]张向颖,任锋,魏琳琳,等.糖原合成酶激酶3β在乙型重型肝炎肝衰竭中的作用研究[J].中华肝脏病杂志,2016,24(4):265-269.
[29]Ren F,Zhang L,Zhang X,et al.Inhibition of glycogen synthase kinase 3βpromotes autophagy to protect mice from acute liver failure mediated by peroxisome proliferatoractivated receptorα.Cell Death Dis.2016;7:e2151.
[30]杨丙章,任锋,温韬,等.糖原合成酶激酶-3β在D-GalN/LPS诱导的小鼠急性肝衰竭中的作用[J].世界华人消化杂志,2012,20(36):3656-3662.