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Strategic Study of CAE >> 2004, Volume 6, Issue 7

Studies of the Guiyuan Fang and Autologous Bone Marrow Stem Cells Transplantation on Hepatic Injury and Fibrosis

1. College of Pharmaceutical Science, Zhejiang University, Hangzhou 310031, China

2. China Academy of Traditional Chinese Medicine, Xiyuan Hospital, Beijing 100091, China

Funding project:国家自然科学基金资助项目(30271663) Received: 2004-03-05 Revised: 2004-04-15 Available online: 2004-07-20

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Abstract

Objective To study the anti-fibrotic and anti-injurious activities of TCM and autologous bone marrow stem cells transplantation. Methods Acute localized liver injury model was induced in rabbits by injecting ethanol locally, and the liver fibrosis models were reproduced by complex factors (high lipid, low protein, alcohol, CCl4). Liver function was assessed by measurement ALT, AST, TP, ALB, and GST. The necrotic areas were measured by pathological analytic system, and the concentrations of serum of PC III , HA in serum were measured by radioimmunoassay. Histopathological changes and degrees of fibrosis, hydroxyproline content in the hepatic tissues were assessed also. /mPA, CK-18, and BrdU - labeled transplanted cells were determined by immunohistochemical staining. Results In the acute liver injury experiment, the reduction of serum GST levels was remarkable in all treated groups, compared with the model group (P<0.05 ). The necrotic regions of the treated groups decrease significantly, compared to model group ( P<0.01). In the inflammatory regions of transplanted rabbits many Brdu-labeled cells were easily observed, most transplanted cells were predominantly in clusters of 1~3 cells each. Part of the cells exhibit the morphology of hepatocyte or bile duct epithelial cells. During liver fibrous experiment, the serum ALT and AST values were reduced significantly in the treated rats (P<0.01), serum TP, ALB, A/G levels were increased in all animals receiving treatment (P<0.01 or 0.05), the serum PC ID , HA levels were also reduced remarkably (P<0.01). Determination of tissue hydroxyproline content confirmed the histological studies, showing a net reduction in total collagen levels. The normal liver tissues showed very weak /xPA expression, and Guiyuan Fang caused a marked increase of /uPA expression in the livers in a dose-dependent manner. The fiPA expression in the autologous BMSCs transplantation group and Guiyuan Fang plus autologous BMSCs transplantation group were the strongest. Double immunohistochemical staining showed the BrdU-labeled cells were fully integrated in the liver parenchyma, along with expression of cytokeratin-18,the liver epithelial specific marker. Some BMSCs have differentiated into hepatocytes or endothelial cells. Conclusion The bone marrow stem cells survive, proliferate, and differentiate in the fibrotic and injurious environment. TCM synergize with bone marrow stem cells to improve liver function, to protect hepatocytes from toxicant, to promote protein synthesis, and to provide a good environment for hepatocyte function. These findings establish the basis for further analysis of BMSCs transplantation in acute and chronic liver diseases.

References

[ 1 ] 李 颖, 彭 仁.阿魏酸钠和当归醇沉物对免疫性肝损伤的干预作用[J].中草药, 2000, 31 (4) :274~276 link1

[ 2 ] 姚希贤, 李校天, 李迎武, 等.丹参等活血化瘀中药对门脉高压动力学影响的临床与实验研究[J].中华消化杂志, 1998, 18 (1) :24~27 link1

[ 3 ] JamallIS , FinelliVN , QueHeeSS .Asimplemethodtodeterminenanogramlevelsof4 hydroxyprolineinbiologicaltissues[J].AnalyticalBiochemistry, 1981, 112 (1) :70~75

[ 4 ] 陈 奇.中药药理研究方法学[M ].北京:人民卫生出版社, 1993.631~632 link1

[ 5 ] MagaudJP , SargentI, JaneClarkeP , etal.DoubleImmunocytochemicallabelingofcellandtissuesampleswithmonoclonalantibromodeoxyuridine[J ].JHistochemCytochem, 1989, 37 (10) :1517~1527

[ 6 ] BruntEM .Gradingandstagingthehistopathologicallesionsofchronichepatitis:TheKnodellhistologyactivityindexandbeyond[J].Hepatology, 2000, 31 (1) :241~246

[ 7 ] 曾民德, 王泰龄, 王宝恩.肝纤维化诊断及疗效评估共识[J].肝脏, 2002, 7 (2) :附3~4 link1

[ 8 ] 王宝恩, 王惠吉, 朱家璇, 等.复方丹参不同剂型治疗肝纤维化实验研究与临床观察[J].中华肝脏病杂志, 1993, 1:69~72

[ 9 ] JiangY , JahagirdarBN , ReinhardtRL , etal.Pluripotencyofmesenchymalstemcellsderivedfromadultmarrow[J].Nature, 2002, 418 (6893) :41~49

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