Figures & data
Figure 1 AS-IV improves STZ-induced INS-1 cell viability. (A) The chemical structure of astragaloside IV. (B and C) Cell viability was detected by Cell Counting Kit-8 assay. The normal INS-1 cells without STZ and AS-IV treatment were served as control. ***P<0.001 vs control; #P<0.05, ##P<0.01 vs STZ+ AS-IV.
![Figure 1 AS-IV improves STZ-induced INS-1 cell viability. (A) The chemical structure of astragaloside IV. (B and C) Cell viability was detected by Cell Counting Kit-8 assay. The normal INS-1 cells without STZ and AS-IV treatment were served as control. ***P<0.001 vs control; #P<0.05, ##P<0.01 vs STZ+ AS-IV.](/cms/asset/82aec9b4-99fa-4d7e-92cf-c8e92d08de3e/dmso_a_347650_f0001_c.jpg)
Figure 2 AS-IV inhibits STZ-induced apoptosis of INS-1 cells. (A) Cell apoptosis was evaluated using a TUNEL assay and (B) quantified. (C) Expression levels of apoptosis-related proteins, Bcl-2, Bax, cleaved caspase 3 and caspase 3 were determined using Western blotting and semi-quantified. β-actin was served as loading control. The normal INS-1 cells without STZ and AS-IV treatment were served as control. ***P<0.001 vs control; #P<0.05, ##P<0.01, ###P<0.001 vs STZ+ AS-IV.
![Figure 2 AS-IV inhibits STZ-induced apoptosis of INS-1 cells. (A) Cell apoptosis was evaluated using a TUNEL assay and (B) quantified. (C) Expression levels of apoptosis-related proteins, Bcl-2, Bax, cleaved caspase 3 and caspase 3 were determined using Western blotting and semi-quantified. β-actin was served as loading control. The normal INS-1 cells without STZ and AS-IV treatment were served as control. ***P<0.001 vs control; #P<0.05, ##P<0.01, ###P<0.001 vs STZ+ AS-IV.](/cms/asset/0d0d9c1b-6e66-4c7a-b2bb-1f9024bf7d7f/dmso_a_347650_f0002_c.jpg)
Figure 3 AS-IV reduces STZ-induced oxidative stress in INS-1 cells. (A) Lipid peroxidation, (B) superoxide dismutase activity and (C) GSH/GSSG Ratio were determined using corresponding kit. The normal INS-1 cells without STZ and AS-IV treatment were served as control. ***P<0.001 vs control; #P<0.05, ##P<0.01, ###P<0.001 vs STZ+ AS-IV.
![Figure 3 AS-IV reduces STZ-induced oxidative stress in INS-1 cells. (A) Lipid peroxidation, (B) superoxide dismutase activity and (C) GSH/GSSG Ratio were determined using corresponding kit. The normal INS-1 cells without STZ and AS-IV treatment were served as control. ***P<0.001 vs control; #P<0.05, ##P<0.01, ###P<0.001 vs STZ+ AS-IV.](/cms/asset/ea095b9e-6d85-4eaa-9b98-c2f0b9bb4aa8/dmso_a_347650_f0003_c.jpg)
Figure 4 AS-IV improves STZ-induced insulin secretion in INS-1 cells. (A) The levels of insulin secretion were detected using the corresponding ELISA kit. Relative mRNA levels of (B) insulin 1 and (C) insulin 2 were determined via RT-qPCR. The normal INS-1 cells without STZ and AS-IV treatment were served as control. ***P<0.001 vs control; #P<0.05, ##P<0.01, ###P<0.001 vs STZ+ AS-IV.
![Figure 4 AS-IV improves STZ-induced insulin secretion in INS-1 cells. (A) The levels of insulin secretion were detected using the corresponding ELISA kit. Relative mRNA levels of (B) insulin 1 and (C) insulin 2 were determined via RT-qPCR. The normal INS-1 cells without STZ and AS-IV treatment were served as control. ***P<0.001 vs control; #P<0.05, ##P<0.01, ###P<0.001 vs STZ+ AS-IV.](/cms/asset/fe8beb07-dc07-4a7a-9513-cf6d9802be6d/dmso_a_347650_f0004_c.jpg)
Figure 5 AS-IV regulates SIRT1/p53 and Akt/GSK3β/Nrf2 signaling pathways. Expression levels of pathway-related proteins, SIRT1, p53, p-Akt, Akt, p-GSK3β, GSK3β and Nrf2, were determined using Western blotting and semi-quantified. β-actin was served as loading control. The normal INS-1 cells without STZ and AS-IV treatment were served as Control. ***P<0.001 vs control; ##P<0.01, ###P<0.001 vs STZ+ AS-IV.
![Figure 5 AS-IV regulates SIRT1/p53 and Akt/GSK3β/Nrf2 signaling pathways. Expression levels of pathway-related proteins, SIRT1, p53, p-Akt, Akt, p-GSK3β, GSK3β and Nrf2, were determined using Western blotting and semi-quantified. β-actin was served as loading control. The normal INS-1 cells without STZ and AS-IV treatment were served as Control. ***P<0.001 vs control; ##P<0.01, ###P<0.001 vs STZ+ AS-IV.](/cms/asset/097fa104-d27b-4882-8750-ea9104a2eb70/dmso_a_347650_f0005_c.jpg)
Figure 6 EX527 (SIRT1 inhibitor) or MK2206 (Akt inhibitor) reversed the protective effect of AS-IV on STZ-induced INS-1 cells. (A) Cell apoptosis was evaluated using a TUNEL assay and (B) quantified. (C) Cell viability was detected by Cell Counting Kit-8 assay. (D) Expression levels of apoptosis-related proteins, Bcl-2, Bax, cleaved caspase 3 and caspase 3 were determined using Western blotting and semi-quantified. β-actin was served as loading control. (E) Lipid peroxidation, superoxide dismutase activity and GSH/GSSG ratio were determined using corresponding kit. (F) The levels of insulin secretion were detected using the corresponding ELISA kit, and the relative mRNA levels of (G) insulin 1 and (H) insulin 2 were determined via RT-qPCR. The normal INS-1 cells without STZ, AS-IV, EX527 and MK2206 treatment were served as Control. ***P<0.001 vs Control; #P<0.05, ##P<0.01, ###P<0.001 vs STZ+ AS-IV+.
![Figure 6 EX527 (SIRT1 inhibitor) or MK2206 (Akt inhibitor) reversed the protective effect of AS-IV on STZ-induced INS-1 cells. (A) Cell apoptosis was evaluated using a TUNEL assay and (B) quantified. (C) Cell viability was detected by Cell Counting Kit-8 assay. (D) Expression levels of apoptosis-related proteins, Bcl-2, Bax, cleaved caspase 3 and caspase 3 were determined using Western blotting and semi-quantified. β-actin was served as loading control. (E) Lipid peroxidation, superoxide dismutase activity and GSH/GSSG ratio were determined using corresponding kit. (F) The levels of insulin secretion were detected using the corresponding ELISA kit, and the relative mRNA levels of (G) insulin 1 and (H) insulin 2 were determined via RT-qPCR. The normal INS-1 cells without STZ, AS-IV, EX527 and MK2206 treatment were served as Control. ***P<0.001 vs Control; #P<0.05, ##P<0.01, ###P<0.001 vs STZ+ AS-IV+.](/cms/asset/80ced3f2-69b2-4db8-8b86-34d9d66ae613/dmso_a_347650_f0006_c.jpg)