ABSTRACT
<p><b>OBJECTIVE</b>To explore effect of interleukin 10 (IL-10) gene-modified bone marrow-derived liver stem cells (BDLSCs) transplantation on hepatic inflammatory response and liver regeneration in rats with liver fibrosis.</p><p><b>METHODS</b>50 female Wistar rats were randomly divided into 4 groups: (1) control group: 10 rats were subcutaneously injected with olive oil for 8 weeks; (2) fibrosis groups: 16 rats were subcutaneously injected with carbon tetrachloride (CCl4) for 8 weeks to induce liver fibrosis; (3) BDLSC group: 12 rats were subcutaneously injected with CCl4 for 8 weeks, and were transplanted with 2 x 10(5) BDLSC at week 4; (4) BDLSC/IL-10 group: 12 rats were subcutaneously injected with CCl4 for 8 weeks, and were transplanted with 2 x 10(5) IL-10 gene-modified BDLSC at week 4. IL-10 and tumor necrosis factor alpha (TNFa) in liver tissues were detected by ELISA. HE stained liver tissues were observed under light microscope. The expression of hepatocyte growth factor (HGF) was quantified by real-time RT-PCR, and the expression of proliferating cell nuclear antigen (PCNA) was determined by immunohistochemistry.</p><p><b>RESULTS</b>The ratio of IL-10/TNFa in fibrosis group (0.05+/-0.01) was lower than that in control group (0.26+/-0.04) (P < 0.01). Transplantation of untreated BDLSCs did not improve the ratio (P > 0.05), however, transplantation of IL-10 modified BDLSCs improved the ratio significantly (P < 0.01). Severe inflammatory response and fibrosis were observed in fibrosis group. Inflammatory response was alleviated to some extent in the BDLSC group, and the histopathology of BDLSC/IL-10 group was not significantly different from that of the control group. Compared to the control group, the expression of HGF mRNA and PCNA protein was increased in the fibrosis group (P < 0.01). The expression of HGF and PCNA was further increased by BDLSCs or IL-10 modified BDLSCs transplantation. Compared to BDLSCs, IL-10 gene-modified BDLSCs were more potent to induce the expression of HGF and PCNA.</p><p><b>CONCLUSION</b>Transplantation of IL-10 gene-modified BDLSCs can alleviate hepatic inflammatory response and promote liver regeneration in hepatic fibrosis rats.</p>
Subject(s)
Animals , Female , Rats , Adenoviridae , Genetics , Bone Marrow Cells , Cell Biology , Cell Proliferation , Disease Models, Animal , Genetic Therapy , Methods , Hepatocyte Growth Factor , Genetics , Metabolism , Immunohistochemistry , Interleukin-10 , Genetics , Liver , Metabolism , Pathology , Liver Cirrhosis , Genetics , Pathology , Therapeutics , Liver Regeneration , RNA, Messenger , Genetics , Metabolism , Rats, Wistar , Reverse Transcriptase Polymerase Chain Reaction , Stem Cell Transplantation , Methods , Transduction, Genetic , Tumor Necrosis Factor-alpha , MetabolismABSTRACT
Objective To study the effects of N-acetyl-seryl-aspartyl-lysyl-proline(AcSDKP) on proliferation and synthesis and secretion of extracellular matrix(ECM) in rat active hepatic stellate cells(HSCs). Methods Primary cultures of HSCs when actived were subjected to AcSDKP(0.01, 0.1, 1, 10 and 100 nmol/L, respectively) treatment, and control group was established. The expression of collagen Ⅰ(ColⅠ) and collagen Ⅲ(Col Ⅲ) mRNA were analyzed with RT-PCR. The proliferation of HSCs was detected by MTT. Hyaluronic acid (HA) and laminin(LN) in the supernatant were determined by enzyme-linked immunosorbent assay. Results Compared with control group, the proliferation of active HSCs was significantly inhibited by 1 and 10 nmol/L AcSDKP. 1, 10 nmol/L AcSDKP and 0.1 to 100 nmol/L AcSDKP significantly inhibited the expression of HSCs ColⅠ mRNA and Col Ⅲ mRNA, respectively. Expression of HA and LN in the supernatant were significantly inhibited by 0.1, 1, 10 nmol/L and 0.1, 1 nmol/L AcSDKP, respectively. ConclusionAcSDKP can inhibit synthesis and secretion of ECM in active HSCs in a dose-dependent manner, with a maximum inhibition effect at 1 nmol/L AcSDKP. The mechanism may involve the inhibition of the proliferation of HSCs,which leads to the decrease of HSCs that synthesize and secrete ECM.
ABSTRACT
Objective To explore the in vitro isolation of ?2m-/Thy-1+ bone marrow-derived liver stem cells(BDLSCs) which bear double features of stem and liver cells from bone marrow stem cells(BMSCs)as so to provide suitable donor cells for the treatment of liver diseases by cellular transplant. Methods ?2m-/Thy-1+ BDLSCs were isolated by magnetic bead cell sorting(MACS) method from cholestatic rats in vitro,and cell purity was detected using flow cytometry.Liver associated phenotype markers were characterized by RT-PCR and immunofluorescence staining. Results BDLSCs isolated by MACS were purified and viable,and possessed hepatocyte-like features at gene and protein levels. Conclusion ?2m-/Thy-1+ BDLSCs are special subsets of BMSCs which may have promising potentials in the stem cell-based treatment of liver diseases.