Effect of Mesenchymal Stem Cells With/Without Silymarin Against Liver Fibrosis in In Vitro Cell Line HepG2 with Study on Nuclear Factor-Kappa β, Caspase3, Apoptosis, Necrosis, Proliferation,and Collagen
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We hypothesize Mesenchymal Stem Cells could Improve Liver Fibrosis. This research is an experimental pre and post test in vitro. Hepatocyte stem cells were taken from the umbilical cord of newborns, processed in the Prodia stem cell laboratory (Prostem). Hep.G2 cell line culture cells from ATCC, USA. Cell lines were divided into 4 groups: Cb (negative control), C1 (positive control / silymarin), C2 (Mesenchymal stem cells / MSC), C3 (MSC+Sylimarin), each group made 3 plates, consisting of 7 wells. Examination was carried out on NF-kβ, Caspase-3, Apoptosis, Necrosis, Proliferation, and cytopathologically seen the growth of collagen. Cytopathological test of cell line hepG2 showed fibroblast growth. In the isolation of UC-MSC, the phenotype results were CD 73+, CD90+, CD105+ and Lin- and condotioned medium BDNF, SDF1, FGF, VEGF, EGF, NGF, IL10, IL1β, IFN-α, MCP1, IL6, IL12p70, IL17A, IL18, IL23 NF-kβ test used Anova test, NF-kβ inhibition at 24 and 48 hours compared to control, significant p<0.05. The difference between the treatment variables at 24 hours was not significant but the difference was significant at 48 hours. Caspase-3 test at 24 hours used Anova test. At 48 hours Kruskal Wallis test. Caspase inhibition compared to control, not significant, between treatment variables significantly different at 48 hours p<0.008 Apoptosis test at 24 hours used the Kruscal Wallis test, while at 48 hours the ANOVA test was used. Apoptotic inhibition compared to control was significant, at 24 hours with p<0.002 at 48 hours with p<0.001. Significant differences in treatment variables were obtained at the 24th hour with p<0.002. While the 48th hour is not significant. Necrosis test at 24 hours used the Kruscal Wallis test and at 48 hours the ANOVA test was used. Necrosis inhibition compared to control, C2 significance, at 24 hours or 48 hours, for C1 and C3 was not significant. The sylimarin test is not significant The proliferation test at 24 and 48 hours used the Kruscal Wallis test. C2 activated proliferation compared to control, significant, p=0.338 at 24 hours and p=0.337 at 48 hours. C1 p=0.002 and C3 p=0.002 at 24 and 48 hours, not significant. The sylimarin test is not significant Our conclusion are disclosed as follow : There is collagen growth. Mesenchymal stem cells inhibited NF-kβ, necrosis, apoptosis, proliferation activation significantly, caspase-3 inhibition was not significant, while sylimarin for NF-kβ inhibitory effect, apoptosis, was significant, but the effect of inhibition of necrosis, caspase and proliferation activation, was not significant.
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This research is an experimental pre and post test in vitro. Hepatocyte stem cells were taken from the umbilical cord of newborns, processed in the Prodia stem cell laboratory (Prostem). Hep.G2 cell line culture cells from ATCC, USA. Cell lines were divided into 4 groups: Cb (negative control), C1 (positive control / silymarin), C2 (Mesenchymal stem cells / MSC), C3 (MSC+Sylimarin), each group made 3 plates, consisting of 7 wells. Examination was carried out on NF-kβ, Caspase-3, Apoptosis, Necrosis, Proliferation, and cytopathologically seen the growth of collagen