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[Induction of peripheral blood mononuclear cells to hepatocyte-like cells and preliminary study of cell response to injury under the effect of acetaminophen].
Liu, P; Zhang, B; Zeng, Q; Chen, S W; Ge, C; Wang, W H; Wang, C Z; Yue, W; Wan, J.
Afiliación
  • Liu P; Chinese LPA Medical School, Beijing 100853, China Department of Gastroenterology, the Second Medical Centre, Chinese PLA General Hospital, National Clinical Research Center for Geriatic Diseases, Beijing 100853, China.
  • Zhang B; Stem Cell and Regenerative Medicine Lab, Institute of Health Service and Transfusion Medicine, Beijing 100850, China.
  • Zeng Q; Stem Cell and Regenerative Medicine Lab, Institute of Health Service and Transfusion Medicine, Beijing 100850, China.
  • Chen SW; Department of Gastroenterology, the Second Medical Centre, Chinese PLA General Hospital, National Clinical Research Center for Geriatic Diseases, Beijing 100853, China.
  • Ge C; Beijing University of Technoloby, Beijing 100124, China.
  • Wang WH; Department of Gastroenterology, the Second Medical Centre, Chinese PLA General Hospital, National Clinical Research Center for Geriatic Diseases, Beijing 100853, China.
  • Wang CZ; Department of Gastroenterology, the Second Medical Centre, Chinese PLA General Hospital, National Clinical Research Center for Geriatic Diseases, Beijing 100853, China.
  • Yue W; Stem Cell and Regenerative Medicine Lab, Institute of Health Service and Transfusion Medicine, Beijing 100850, China.
  • Wan J; Department of Gastroenterology, the Second Medical Centre, Chinese PLA General Hospital, National Clinical Research Center for Geriatic Diseases, Beijing 100853, China.
Zhonghua Gan Zang Bing Za Zhi ; 30(1): 87-93, 2022 Jan 20.
Article en Zh | MEDLINE | ID: mdl-35152675
Objective: To establish a method for the induction of peripheral blood mononuclear cells to hepatocyte-like cells, and preliminarily investigate cell response to injury under the effect of acetaminophen (APAP). Methods: The surface marker CD45 of peripheral blood mononuclear cells wase detected cells by using flow cytometry and immunofluorescence methods. The cellular morphology of induced hepatocyte-like cells was observed under an inverted microscope. Real-time fluorescent quantitative PCR (RT-PCR) was used to detect the expression level of hepatocyte-specific genes, such as cytochrome (CY) P1A2, CYP3A4, CYP2C9, albumin (ALB), alpha-fetoprotein (AFP), and hepatocyte nuclear factor (HNF)4α mRNA. Immunofluorescence method was used to detect intracellular hepatocyte markers AFP, HNF4α, and ALB expression at the protein level. Biochemical analyzer was used to detect hepatocyte-specific secretory functions of AFP, ALB, and urea. Luciferase chemiluminescence method was used to detect the activity of key drug metabolizing enzyme CYP3A4. Colorimetric assay was used to detect the effect of the drug acetaminophen on hepatocyte-like cells, and alanine aminotransferase (ALT) was used as an indicator of liver cell injury. The statistical differences between the data were compared with t-test and rank-sum test. Results: The positive expression rate of CD45 cell surface markers isolated from peripheral blood mononuclear cells was about 98%, and hepatocyte-like cell morphology changes appeared on 15th day of induction. Compared with isolated mononuclear cells, CYP1A2, CYP3A4, CYP2C9, ALB, AFP and HNF4α mRNA was markedly elevated. The expression level of AFP, ALB and HNF4α protein were equally increased, and the secretory function of AFP, ALB and urea were enhanced. Compared with primary hepatocytes, CYP1A2, CYP2C9, AFP, HNF4α mRNA, and CYP3A4 mRNA did not decrease. The expression levels of AFP, ALB, and HNF4α proteins in the cells did not decrease, and the secretory function of AFP, ALB, and urea did not decrease. In addition, the CYP3A4 enzyme activity produced by hepatocyte-like cells was similar to that of primary hepatocytes. Compared with hepatocyte-like cells incubated without APAP, hepatocyte-like cells incubated with APAP had higher ALT level. Under the effect of APAP, the ALT level of hepatocyte-like cells was higher than isolated mononuclear cells. Conclusion: Peripheral blood mononuclear cells can be induced into hepatocyte-like cells with partial characteristics of hepatocytes, including the activity of CYP3A4, a key enzyme of hepatocyte drug metabolism. Additionally, preliminarily ALT secretory features reflect the hepatocytes injury under the effect of acetaminophen.
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Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Leucocitos Mononucleares / Acetaminofén Idioma: Zh Revista: Zhonghua Gan Zang Bing Za Zhi Asunto de la revista: GASTROENTEROLOGIA Año: 2022 Tipo del documento: Article País de afiliación: China

Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Leucocitos Mononucleares / Acetaminofén Idioma: Zh Revista: Zhonghua Gan Zang Bing Za Zhi Asunto de la revista: GASTROENTEROLOGIA Año: 2022 Tipo del documento: Article País de afiliación: China