The Protective Effect Of Trichostatin A And5-Azacitidine On Pancreatic β-cells

Objective To investigate the protective effect of histone deacetylase inhibitor Trichostatin A and DNA methyltransferases inhibitor5-Azacitidine on cytokine-induced toxicity in pancreatic β-cells.Methods Apoptosis was induced in vitro by interleukin-1β(IL-1β) and interferon-γ(IFN-γ).After treatment with Trichostatin A(TSA) at0.05umol/L,0.1umol/L respectively or5-Azacitidine (5-AzaC) at0.63umol/L,1.25umol/L respectively or Trichostatin A at0.1umol/L combined with5-Azactidine at1.25umol/L, the viability of RIN-m5f cells was detected by MTT experiment. Apoptotic rate of RTN-m5f cells was determined by Annexin V-FITC/PI flow cytometry. Insulin secretion measured by ELISA.Results After treatment of different concentrations of TSA/5-AzaC, the viability of RIN-m5f cells increased to79.2%,82.9%,89.1%respectively. The apoptotic rate of RIN-m5f cells decreased to8.2%,10.5%,7.9%respectively. There were significant differences in viability and apoptotic rate between the TSA/5-AzaC treatment group and IL-1β/IFN-y group (33.9%,16.6%, P<0.05). After treatment with TSA/5-AzaC, glucose-stimulated insulin secretion (GSIS) of RIN-m5f cells recovered in different degrees [(5.53±0.16) ng/ml,(5.70±0.18) ng/ml,(5.90±0.10) ng/ml, P<0.05]. There were significant differences in GSIS between the TSA/5-AzaC treatment group and IL-1β/IFN-γ group [(3.21±0.17) ng/ml, P<0.05].Conclusion TSA and5-AzaC might protect pancreatic β-cells directly via inhibiting cytokine-induced apoptosis and recovering insulin secretion. Objective To investigate the protective effect of histone deacetylase inhibitor Trichostatin A and DNA methyltransferases inhibitor5-Azacitidine on high glucose-induced toxicity in pancreatic (3-cells.Methods Apoptosis was induced in vitro by high glucose (33.3mmol/L). After treatment with Trichostatin A(TSA) at0.05umol/L,0.1umol/L respectively or5-Azacitidine (5-AzaC) at0.63umol/L,1.25umol/L respectively or Trichostatin A at0.1umol/L combined with5-Azacitidine at1.25umol/L. The viability of RIN-m5f cells was detected by MTT experiment. Apoptotic rate of RIN-m5f cells was determined by Annexin V-FITC/PI flow cytometry. Insulin secretion measured by ELISA.Results After treatment of different concentrations of TSA/5-AzaC, the viability of RIN-m5f cells increased to79.2%,81.8%,87.5%respectively. The apoptotic rate of RIN-m5f cells decreased to10.4%,9.9%,8.7%respectively.There were significant differences in viability and apoptotic rate between the TSA/5-AzaC treatment group and high glucose group (63.2%,15.8%, P<0.05). After treatment with TSA/5-AzaC, glucose-stimulated insulin secretion (GSIS) of RIN-m5f cells recovered in different degrees [(5.59±0.21) ng/ml,(5.80±0.16) ng/ml,(6.05±0.11) ng/ml, P<0.05]. There were significant differences in GSIS between the TSA/5-AzaC treatment group and high glucose group [(3.37±0.12) ng/ml, P<0.05].Conclusion TSA and5-AzaC might protect pancreatic β-cells directly via inhibiting high glucose-induced apoptosis and recovering insulin secretion.