In Vivo Effect of Vitamin C on Mesenchymal Stem Cell Hepatogenesis After CCl4 Induced Hepatic Fibrosis in Mice

Mai Khaled Abd El-Monaem


Abstract


T his study was done at Medical Biochemistry Department, Ain Shams University Faculty of Medicine during the period from 2014 – 2015 and included 7 groups of albino mice. The aim of this study to assess the effect of daily vitamin C supplementation on labeled bone marrow mesenchymal stem cell of CCL4 induced injured liver and evaluate its effect on stem cell ability to regenerate functioning hepatocytes through comparative measuring of liver functions (serum ALT, AST, and serum albumin levels) as well as relative quantitative assessment of gene expression to the followings (albumin & matrix metalloproteinase- 2 {MMP-2} genes) using real time PCR. The studied groups of mice were classified into 7 main groups as follow: 1. Group 1 (control): injected IP (intra-peritoneal) with normal saline a dose of 0.8 ml/kg once weekly for three successive weeks then sacrificed three weeks later after last injection. 2. Group 2 (CCL4): injected IP (intra-peritoneal) with CCL4 a dose of 0.8 ml/kg once weekly for three successive weeks then sacrificed three weeks later after last injection. 3. Group 3(A): injected IP (intra-peritoneal) with CCL4 a dose of 0.8 ml/kg once weekly for three successive weeks followed one week later by vitamin C (IP) injection at a dose of 2 gm/kg/day for three successive weeks daily then sacrificed after last injection. 4. Group 3(B): injected IP (intra-peritoneal) with CCL4 a dose of 0.8 ml/kg once weekly for three successive weeks followed 1 week later from last injection by vitamin C IP injection at a dose of 4 gm/kg/day for three successive weeks then sacrificed after last injection. 5. Group 4: injected IP (intra-peritoneal) with CCL4 a dose of 0.8 ml/kg once weekly for three successive weeks followed one week later from last injection by IV (intravenous) injection of labeled BM-MSCs with PKH-26 red fluorescent dye at a dose of (1x106) cells once at tail then sacrificed three weeks later. 6. Group 5(A): injected IP (intra-peritoneal) with CCL4 a dose of 0.8 ml/kg once weekly for three successive weeks followed one week later by both IV labeled BM-MSCs with PKH-26 red fluorescent dye at a dose of (1x106) cells once at tail & daily IP vitamin C injection at a dose of 2 gm/kg/day for three successive weeks then sacrificed after last injection. 7. Group 5(B): injected IP (intra-peritoneal) with CCL4 a dose of 0.8 ml/kg once weekly for three successive weeks followed one week later from last dose by both IV labeled BM-MSCs with PKH-26 red fluorescent dye at a dose of (1x106) cells once at tail & daily IP vitamin C injection at a dose of 4 gm/kg/day for three successive weeks then sacrificed after last injection. All groups were subjected to  Estimation of serum ALT, AST & albumin.  Tissue MDA (malondialdehyde) estimation in liver tissue.  Evaluation of both MMP-2(matrix metalloproteinase-2) gene & albumin gene expressions by relative quantification REAL TIME PCR.  Evaluation of labeled stem cells homing using fluorescent microscope for detection (group4, group5A&5B).  Histo-pathological examination of H&E stained liver tissue using light microscope.  Histo-pathological fibrosis evaluation of Masson trichromate stained liver tissue using light microscope.


Other data

Other Titles دراسة تأثير فيتامين ج على الخلايا الجذعية المستخدمة فى بناء خلايا كبد جديدة بعد إصابتها بمركب كربون رباعى الكلوريد فى فئران التجارب
Issue Date 2016
URI http://research.asu.edu.eg/handle/12345678/2718


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