Application Of Raman Spectroscopy In NIT-1 Cells And The Protein Tuberin Cells Induced By High Glucose

Objective:To investigate the role of rosiglitazone in different concentrations of glucose-induced protein tuberin ofβcells, and explore the effects of protein tuberin on the function and proliferation,apoptosis of pancreaticβ-cell.Methods:The NIT-1 cells were puted by 5×104 cells/hole in 24-holes culture plate, and cultured for 48 hours later were random accessed to the treatment group 5.6 group (containing glucose 5.6mmol/L),7.8 group (7.8mmol/L),11.1 group (11.1 mmol/L),16.7 Group (16.7mmol/L),22.2 Group (22.2mmol/L) and 27.6 group (27.6mmol/L), and then 24 hours later were respectively intervented by 10"5 mmol/L rosiglitazone for 48h. Single NIT-1 cell was measured by Raman spectroscopy, the expression in protein lever of p-tuberin by Western blot in each group at the time of 48h intervention.Results:(1) At the high concentration of glucose, NIT-1 cells were tested by Raman spectroscopy,The most noticeable changes were the decrease of spectral intensity at the788cm-1,1002 cm-1,1085 cm-1,1445 cm-1 and 1655 cm-1 peaks, mainly corresponding to protein and lipid, and the peaks were gradually decreased along with increasing glucose concentration(every group compared with 11.1 group P<0.05),11.1 group>16.7 group>22.5 group.(2) After the intervention of 10-5mol/1 rosiglitazone, the peaks mationed had showed no significant alterations. the peaks of none rosiglitazone intervention groups were higher than that of rosiglitazone intervention groups.(3) The expression in tuberin was lowest at 5.6mmol/L glucose concentration(every group compared with 11.1 groups P<0.05), and tuberin increased along with the glucose's level. The expression of tuberin of the 5.6mmol/L group was the lowest,less than any other group(P<0.05); The 27.6mmol/L group had showed no significant alterations with the 22.5 mmol/L group(P>0.05) while significantly higher than the 5.6mmol/L and 11.1mmol/ L,16.7mmol/L group (P<0.05).(4) After the intervention of 10-5mol/1 rosiglitazone,the expression of tuberin had the same trend that was still 5.6 Group<11.1 group<16.7 group <22.5 group<27.6 group, the 27.6mmol/L group was significantly higher than the other group (P<0.05);The expression of overall tuberin was decreased significantly than non-intervention group. At the same glucose concentration, The expression of tuberin of none rosiglitazone intervention groups were higher than that of rosiglitazone intervention group s(inter-group difference was significant P<0.05). Conclusion:High concentration of glucose could hinder beta cell proliferation, induce islet cell apoptosis, and reduce insulin secretion, raise protein tuberin. Above it showed that the islet cell apoptosis,insulin resistance caused by high concentration of glucose may be related with the change of tuberin.Rosiglitazone could improve the function and proliferation, and inhibit the apoptosis of pancreaticβcells, through directly regulating the protein tuberin. This prompted that the measures to regulate the expression of TSC2 can effectively influence the secretion,proliferation,apoptosis of pancreatic (3 cells.