Effects Of Ganoderma Lucidum Proteoglycan On Apoptosis Of ?-cell Induced By Human Amylin

Objective:Early animal experiments confirmed that Ganoderma lucidum extract Ganoderma lucidum polysaccharide(FYGL)has hypoglycemic effect,This study further explored whether FYGL has a protective effect on human amylin-induced apoptosis of pancreatic islet ? cells.Methods:(1)Toxicity test of FYGL on normal INS-1 islet cells:Prepare different concentrations of FYGL(1ug/ml,5ug/ml,10ug/ml,50ug/ml,100ug/ml,500ug/ml,1000ug/ml,1500ug/ml,2000ug/ml),The INS-1 cell line was cultured and inoculated into a 96-well plate.After the cells were attached,the different doses of FYGL were added to intervene for 24 hours.XTT assay was used to detect the activity of each group.(2)Effect of FYGL on IAPP-induced INS-1 cell activity: The cells were cultured in 96-well plates and divided into five groups: normal cell group(CON),model group(25u M IAPP)and FYGL dose intervention groups(25u M IAPP +FYGL 10ug/mll,25 u M IAPP +FYGL 50ug/ml,25 u M IAPP +FYGL 100ug/ml)for 24 hours.XTT assay for cell viability.(3)Effect of FYGL on IAPP-induced INS-1 cell activity: The cells were cultured in 24-well plates and divided into five groups: normal cell group(CON),model group(25u M IAPP)and FYGL dose intervention groups(25u M IAPP +FYGL 10ug/mll,25 u M IAPP +FYGL 50ug/ml,25 u M IAPP +FYGL 100ug/ml)for 24 hours.TUNEL assay for cell viability.(4)Effect of FYGL on IAPP-induced apoptosis of INS-1 cells and expression of endoplasmic reticulum stress-related factors: Grouped as above,real-time PCR was used to detect apoptosis and expression of endoplasmic reticulum stress-related factors(caspase3,bcl-2,CHOP,JNK1).Results:(1)None of the doses of FYGL produced toxic effects on normal INS-1 cells.(2)XTT results showed that 25 u MIAPP significantly decreased cell viability compared with CON group(P<0.01).Compared with MOD group,FYGL increased cell viability in a dose-dependent manner.The difference was statistically significant(P<0.05).(3)TUNEL results:Compared with the con group,The apoptotic cells increased significantly in IAPP group.,After FYGL intervention,Apoptotic cells decreased significantly,and 100 ug/ml FYGL was the best.(P<0.01).(5)Real-time PCR results: Compared with the con group,the expression of caspase3,CHOP and JNK1 in the MOD group increased significantly.Compared with the con group,the expression of bcl-2 in the MOD group decreased significantly,and the difference was statistically significant(P <0.01).FYGL could inhibit the expression of caspase3,CHOP and JNK1 induced by IAPP.On the contrary,FYGL could increase the expression of bcl-2 in a dose-dependent manner,and the difference was statistically significant(P<0.01).Conclusion:FYGL inhibited h IAPP-induced apoptosis of INS1 islet cells in a dose-dependent manner.FYGL may inhibit the apoptosis of INS1 cells induced by h IAPP by affecting the expression of endoplasmic reticulum stress CHOP,JNK1 and the expression of apoptotic related factors caspase 3 and bcl-2,possibly as a potential hypoglycemic agent with protective effect on islet ? cells.