人成体肝源性干细胞对酒精性脂肪肝小鼠模型的防治作用
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摘要
目的探讨人肝源性干细胞腹腔移植对小鼠酒精性脂肪肝的防治作用。方法将30只雄性C57BL/6小鼠随机分为空白对照组(N组)、模型对照组(M组)以及肝干细胞移植组(S组)。N组以基础饲料喂养,M组和S组以Lieber-DeCarli酒精液体饲料喂养,S组每周进行2次腹腔肝干细胞移植。干预6周后,测定各组小鼠体质量、肝脏指数,检测血清ALT、AST、TBil、TC、TG、HDL-C、LDL-C,检测肝脏中TG和游离脂肪酸(NEFA)水平,并做肝脏病理学检查和肝组织油红O染色。计量资料多组间比较采用单因素方差分析,进一步两两比较采用SNK-q检验。结果 3组间比较,小鼠血清ALT、AST、TG、TC、HDL-C水平和肝组织TG、NEFA水平差异均具有统计学意义(F值分别为66. 94、7. 15、8. 02、18. 64、3. 86、23. 14、30. 49,P值均<0. 05)。与N组对比,M组小鼠的血清中ALT、AST、TG和肝组织中TG、NEFA明显升高,差异均有统计学意义(P值均<0. 05)。S组小鼠的血清ALT、AST、TG和肝组织TG、NEFA都明显低于M组,差异均有统计学意义(P值均<0. 05);肝脏HE染色和油红O染色均显示S组小鼠的肝脏脂肪变性程度较M组明显减轻。结论腹腔肝源性干细胞移植对小鼠酒精性脂肪肝具有显著的防治作用。
Objective To investigate the effect of intraperitoneal transplantation of human liver-derived stem cells in the prevention and treatment of alcoholic fatty liver disease in mice. Methods A total of 30 male C57 BL/6 mice were randomly divided into blank control group( group N),model control group( group M),and stem cell transplantation group( group S). The mice in group N were fed a normal diet,and those in the other two groups were fed Lieber-DeCarli alcohol liquid diet; at the same time,the mice in group S were given intraperitoneal transplantation of human liver-derived stem cells twice a week. After six weeks of intervention,body weight and liver index were measured,and the serum levels of alanine aminotransferase( ALT),aspartate aminotransferase( AST),total bilirubin( TBil),total cholesterol( TC),triglyceride( TG),high-density lipoprotein cholesterol( HDL-C),and low-density lipoprotein cholesterol( LDL-C) were also measured. The levels of TG and non-esterified fatty acid( NEFA) in the liver were measured,and liver pathological examination and oil red O staining of the liver were performed. One-way ANOVA was used for comparison of continuous data between multiple groups,and the SNK-q test was used for further comparison between two groups. Results There were significant differences in the serum levels of ALT,AST,TG,TC,and HDL-C and the content of TG and NEFA in the liver between the three groups( F = 66. 94,7. 15,8. 02,18. 64,3. 86,23. 14 and 30. 49,all P < 0. 05),Compared with group N,group M showed significant increases in levels of ALT,AST,and TG in serum and levels of TG and NEFA in liver tissue( all P < 0. 05). Group S had significantly lower levels of ALT,AST,and TG in serum and levels of TG and NEFA in liver tissue than group M( all P < 0. 05). Liver HE staining and oil red O staining showed that group S had a significantly lower degree of liver steatosis than group M. Conclusion Intraperitoneal transplantation of human liver-derived stem cells has a marked effect in the prevention and treatment of alcoholic fatty liver disease in mice.
Objective To investigate the effect of intraperitoneal transplantation of human liver-derived stem cells in the prevention and treatment of alcoholic fatty liver disease in mice. Methods A total of 30 male C57 BL/6 mice were randomly divided into blank control group( group N),model control group( group M),and stem cell transplantation group( group S). The mice in group N were fed a normal diet,and those in the other two groups were fed Lieber-DeCarli alcohol liquid diet; at the same time,the mice in group S were given intraperitoneal transplantation of human liver-derived stem cells twice a week. After six weeks of intervention,body weight and liver index were measured,and the serum levels of alanine aminotransferase( ALT),aspartate aminotransferase( AST),total bilirubin( TBil),total cholesterol( TC),triglyceride( TG),high-density lipoprotein cholesterol( HDL-C),and low-density lipoprotein cholesterol( LDL-C) were also measured. The levels of TG and non-esterified fatty acid( NEFA) in the liver were measured,and liver pathological examination and oil red O staining of the liver were performed. One-way ANOVA was used for comparison of continuous data between multiple groups,and the SNK-q test was used for further comparison between two groups. Results There were significant differences in the serum levels of ALT,AST,TG,TC,and HDL-C and the content of TG and NEFA in the liver between the three groups( F = 66. 94,7. 15,8. 02,18. 64,3. 86,23. 14 and 30. 49,all P < 0. 05),Compared with group N,group M showed significant increases in levels of ALT,AST,and TG in serum and levels of TG and NEFA in liver tissue( all P < 0. 05). Group S had significantly lower levels of ALT,AST,and TG in serum and levels of TG and NEFA in liver tissue than group M( all P < 0. 05). Liver HE staining and oil red O staining showed that group S had a significantly lower degree of liver steatosis than group M. Conclusion Intraperitoneal transplantation of human liver-derived stem cells has a marked effect in the prevention and treatment of alcoholic fatty liver disease in mice.
引文
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[13] LOMBARD R,BUZZETTI E,ROCCARINA D,et al. Non-in-vasive assessment of liver fibrosis in patients with alcoholic liv-er disease[J]. World J Gastroenterol,2015,21(39):11044-11052.
[14] MANDAL A,GRUPP A,SCHWEDERSKI B,et al. Noninno-cently behaving bridging anions of the widely distributed an-tioxidant ellagic acid in diruthenium complexes[J]. InorgChem,2015,54(20):10049-10057.
[15] LIN XX,CHEN D,ZHAO Q,et al. Genetic research on the in-cidence of alcoholic liver disease[J]. Chin J Clin PharmacolTher,2017,22(11):1309-1314.(in Chinese)林秀贤,陈丹,赵青,等.影响酒精性肝病发病的遗传学研究进展[J].中国临床药理学与治疗学,2017,22(11):1309-1314.
[16] SZABO G. Gut-liver axis in alcoholic liver disease[J]. Gas-troenterology,2015,148(1):30-36.
[17] DI CAMPLI C,PISCAGLI AC,PIERELLI L,et al. A human um-bilical cord stem cell rescue therapy in a murine model of toxicliver injury[J]. Dig Liver Dis,2004,36(9):603-613.
[18] YANG S,KOTEISH A,LIN H,et al. Oval cells compensate fordamage and replicative senescence of mature hepatocytes inmice with fatty liver disease[J]. Hepatology,2004,39(2):403-411.
[19] MARGINI C,VUKOTIC R,BRODOSI L,et al. Bone marrowderived stem cells for the treatment of end-stage liver dis-ease[J]. World J Gastroenterol,2014,20(27):9098-9105.
[20] CHEN Q,KHOURY M,LIMMON G,et al. Human fetal hepaticprogenitor cells are distinct from,but closely related to,hema-topoietic stem/progenitor cells[J]. Stem Cells,2013,31(6):1160-1169.
[21] PICHARD V,FERRY N. Origin of small hepatocyte-like pro-genitor in retrorsine-treated rats[J]. J Hepatol,2008,48(2):368-369.
[22] GERBAL-CHALOIN S,FUNAKOSHI N,CAILLAUD A,et al.Human induced pluripotent stem cells in hepatology:Beyondthe proof of concept[J]. Am J Pathol,2014,184(2):332-347.
[23] ENOSAWA S,HORIKAWA R,YAMAMOTO A,et al. Hepato-cyte transplantation using a living donor reduced graft in a ba-by with ornithine transcarbamylase deficiency:A novel sourceof hepatocytes[J]. Liver Tranpl,2014,20(3):391-393.
[2] European Association for the Study of Liver. EASL clinical practi-cal guidelines:Management of alcoholic liver disease[J]. J Hep-atol,2012,57(2):399-420.
[3] WARREN KR,MURRAY MM. Alcoholic liver disease and pan-creatitis:Global health problems being addressed by the USNational Institute on Alcohol Abuse and Alcoholism[J]. J Gas-troenterol Hepatol,2013,28(Suppl 1):4-6.
[4] O'SHEA RS,DASARATHY S,MCCULLOUGH AJ. Alcoholicliver disease[J]. Hepatology,2010,51(1):307-328.
[5] MCCULLOUGH AJ,O'SHEA RS,DASARATHY S. Diagnosisand management of alcoholic liver disease[J]. J Dig Dis,2011,12(4):257-262.
[6] FAN JG. Epidemiology of alcoholic and nonalcoholic fatty liverdisease in China[J]. J Gastroenterol Hepatol,2013,28(Sup-pl 1):11-17.
[7] AKIYAMA K,CHEN C,WANG D,et al. Mesenchymal-stem-cell-induced immunoregulation involves FAS-ligand-/FAS-mediated T cell apoptosis[J]. Cell Stem Cell,2012,10(5):544-555.
[8] KUO TK,HUNG SP,CHUANG CH,et al. Stem cell therapyfor liver disease:Parameters governing the success of usingbone marrow mesenchymal stem cells[J]. Gastroenterology,2008,134(7):2111-2121.
[9] ZHANG Z,WANG FS. Stem cell therapies for liver failure andcirrhosis[J]. J Hepatol,2013,59(1):183-185.
[10] SPAHR L,LAMBERT JF,RUBBIA-BRANDT L,et al. Granu-locyte-colony stimulating factor induces proliferation of hepat-ic progenitors in alcoholic steatohepatitis:A randomized trial[J]. Hepatology,2008,48(1):221-229
[11] AURICH H,SGODDA M,KALTWASSER P,et al. Hepatocytedifferentiation of mesenchymal stem cells from human adiposetissue in vitro promotes hepatic integration in vivo[J]. Gut,2009,58(4):570-581.
[12] BI YZ,FAN Z,CHEN DF,et al. Protective effect of intraperito-neal transplantation of human fiver-derived stem cells at dif-ferent times against concanavalin A-induced acute liver injuryin mice[J]. Chin J Hepatol,2017,25(3):205-210.(inChinese)毕研贞,樊增,陈东风,等.人肝源性干细胞腹腔移植不同时间对Con A诱导小鼠急性肝损伤保护作用[J].中华肝脏病杂志,2017,25(3):205-210.
[13] LOMBARD R,BUZZETTI E,ROCCARINA D,et al. Non-in-vasive assessment of liver fibrosis in patients with alcoholic liv-er disease[J]. World J Gastroenterol,2015,21(39):11044-11052.
[14] MANDAL A,GRUPP A,SCHWEDERSKI B,et al. Noninno-cently behaving bridging anions of the widely distributed an-tioxidant ellagic acid in diruthenium complexes[J]. InorgChem,2015,54(20):10049-10057.
[15] LIN XX,CHEN D,ZHAO Q,et al. Genetic research on the in-cidence of alcoholic liver disease[J]. Chin J Clin PharmacolTher,2017,22(11):1309-1314.(in Chinese)林秀贤,陈丹,赵青,等.影响酒精性肝病发病的遗传学研究进展[J].中国临床药理学与治疗学,2017,22(11):1309-1314.
[16] SZABO G. Gut-liver axis in alcoholic liver disease[J]. Gas-troenterology,2015,148(1):30-36.
[17] DI CAMPLI C,PISCAGLI AC,PIERELLI L,et al. A human um-bilical cord stem cell rescue therapy in a murine model of toxicliver injury[J]. Dig Liver Dis,2004,36(9):603-613.
[18] YANG S,KOTEISH A,LIN H,et al. Oval cells compensate fordamage and replicative senescence of mature hepatocytes inmice with fatty liver disease[J]. Hepatology,2004,39(2):403-411.
[19] MARGINI C,VUKOTIC R,BRODOSI L,et al. Bone marrowderived stem cells for the treatment of end-stage liver dis-ease[J]. World J Gastroenterol,2014,20(27):9098-9105.
[20] CHEN Q,KHOURY M,LIMMON G,et al. Human fetal hepaticprogenitor cells are distinct from,but closely related to,hema-topoietic stem/progenitor cells[J]. Stem Cells,2013,31(6):1160-1169.
[21] PICHARD V,FERRY N. Origin of small hepatocyte-like pro-genitor in retrorsine-treated rats[J]. J Hepatol,2008,48(2):368-369.
[22] GERBAL-CHALOIN S,FUNAKOSHI N,CAILLAUD A,et al.Human induced pluripotent stem cells in hepatology:Beyondthe proof of concept[J]. Am J Pathol,2014,184(2):332-347.
[23] ENOSAWA S,HORIKAWA R,YAMAMOTO A,et al. Hepato-cyte transplantation using a living donor reduced graft in a ba-by with ornithine transcarbamylase deficiency:A novel sourceof hepatocytes[J]. Liver Tranpl,2014,20(3):391-393.