Induction of rat liver parenchymal cell apoptosis by hepatic myofibroblasts via transforming growth factor beta |
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| Affiliation: | 1. Hunan Province Key Laboratory of Typical Environmental Pollution and Health Hazards, School of Public Health, Hengyang Medical School, University of South China, Hengyang 421001, PR China;2. Key Discipline Laboratory for National Defense for Biotechnology in Uranium Mining and Hydrometallurgy, University of South China, Hengyang 421001, PR China;1. Laboratório de Genética, Instituto Butantan, São Paulo, São Paulo, Brazil;2. Programa de Pós-graduação Interunidades em Biotecnologia, Instituto de Ciências Biomédicas (ICB), Universidade de São Paulo (USP), São Paulo, São Paulo, Brazil;3. Laboratório de Biologia Molecular, Genética e Mutagênese, Faculdade de Ciências e Letras de Assis, Universidade Estadual Paulista “Júlio de Mesquita Filho” (UNESP), Assis, São Paulo, Brazil;4. Programa de Pós-graduação em Morfologia Funcional e Estrutural, Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP), São Paulo, São Paulo, Brazil;1. Universidade da Coruña, Gerontology and Geriatrics Research Group, Instituto de Investigación Biomédica de A Coruña (INIBIC), Complexo Hospitalario Universitario de A Coruña (CHUAC), Servizo Galego de Saúde (SERGAS), A Coruña, Spain;2. Universidade da Coruña, Grupo NanoToxGen, CICA – Centro Interdisciplinar de Química e Bioloxía, Departamento de Biología, A Coruña, Spain;3. Instituto de Investigación Biomédica de A Coruña (INIBIC), A Coruña, Spain;4. Universidade da Coruña, Grupo DICOMOSA, CICA – Centro Interdisciplinar de Química e Bioloxía, Departamento de Psicología, A Coruña, Spain;5. Department of Environmental Health, Portuguese National Institute of Health, Porto, Portugal;6. EPIUnit—Instituto de Saúde Pública, Universidade do Porto, Porto, Portugal;7. Laboratório para a Investigação Integrativa e Translacional em Saúde Populacional (ITR), Porto, Portugal;1. Universidad Continental, Lima, Peru;2. Oficina de Epidemiología, Hospital Regional Lambayeque, Lambayeque, Peru;3. Grupo de Investigación ACEMED-UPTC, Universidad Pedagógica y Tecnológica de Colombia, Tunja, Colombia;4. Facultad de Ciencias Médicas, Universidad Nacional de Caaguazú, Coronel Oviedo, Paraguay;5. IRCCS Fondazione Don Carlo Gnocchi, Milan, Italy;6. The Medical School, The University of Sheffield, Sheffield, United Kingdom;7. Department of Research Analytics, Saveetha Dental College and Hospitals, Saveetha Institute of Medical and Technical Sciences (SIMATS), Chennai, India;8. Department of Pharmacy and Pharmacology, SRM College of Pharmacy, SRM Institute of Science and Technology, Kattankulathur, Chennai, India;9. Universidad de Huánuco, Huanuco, Peru;1. Department of Occupational and Environment Health, College of Public Health, Zhengzhou University, 450001 Henan, China;2. Prevention and Infection Control Section, Xi''an Union Hospital, 710199 Xi''an, Shaanxi Province, China;3. Department of Ultrasound, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou, 450015 Henan, China;4. Department of Child and Adolescence Health, College of Public Health, Zhengzhou University, 450001 Henan, China;1. School of Public Health Jilin University, Changchun, Jilin 130021, PR China;2. School of Public Health, Capital Medical University, Beijing, PR China;3. Beijing Key Laboratory of Environmental Toxicology, Capital Medical University, Beijing, PR China |
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| Abstract: | The induction of apoptosis of rat liver parenchymal cells (PC) by transforming growth factor beta (TGF-beta)-expressing transforming fat-storing cells (FSC), i.e., myofibroblasts (MFB), was studied under culture conditions and compared with the apoptotic effect of human recombinant TGF-beta1. MFB were obtained by subculture of FSC. The TGF-beta concentration in the conditioned medium of myofibroblast (MFBcM) determined with the Mink cell proliferation inhibition assay was <0.25 ng/mL/24 in the native medium, but 1.9 ng/mL24 h after transient acidification. MFBcM added in various dilutions and for different times to PC monolayers induced progressive cell detachment from the plastic support and increase of lactate dehydrogenase (LDH) activity in the medium. The reduction of mitochondrial dehydrogenase activity in PC (XTT or WST-1 test) was an early sign of MFBcM-induced functional impairment of PC. Short term exposure of PC with MFBcM for 3 hours was sufficient to induce the deleterious effects on PC, but neither native (nonactivated) MFBcM nor conditioned medium of untransformed FSC (FSCcM), in which TGF-beta was not detectable, were able to impair function and viability of PC. Activated MFBcM increased strongly (up to 21-fold) the concentration of oligonucleosomal DNA fragments both in the adherent and detached fraction of PC. Internucleosomal DNA fragments (DNA ladder) were demonstrated by electrophoresis of extracted DNA on agarose gels and by in situ end-labeling of DNA breaks (TUNEL reaction) only in MFBcM-exposed PC. MFBcM-treated PC exhibited intense fluorescence after staining with DNA-binding dye Hoechst 33342 and an increased number of cells with fragmented nuclei. All these criteria point to MFBcM-generated apoptosis of cultured PC, which were found to be very similar to those induced by human recombinant TGF-beta1. The exclusive role of active TGF-beta in MFBcM as mediator of the apoptotic effects of MFB was proven by preincubation of the conditioned medium with human recombinant latency-associated peptide, which reversed completely MFBcM induced reduction of the XTT-test and the MFBcM-generated increase of oligonucleosomal DNA fragments. Partial reversibility was reached by preincubation of the medium with recombinant soluble type II TGF-beta receptor. The data let us conclude that transformed FSC, i.e., MFB in damaged liver, could participate in the mechanisms of PC apoptosis by paracrine loops involving TGF-beta. (Hepatology 1996 Mar;23(3):571-81) |
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