?? 0.01 and ? 0.05 compared with the MOD group. To evaluate the metabolic abilities in diabetic rats, OGTT was performed. exposing that this IRS-2/Akt/GSK-3signaling axis was involved in the protective effects of CHS. These results demonstrate that CHS guarded (GSK-3acts as an important unfavorable regulator and plays a central role in the PI3K/Akt pathway [22]. Knockout of GSK-3in and guarded the brain from oxidative stress [24]. In pancreatic and acquired from the New Drug Research and Development Center, Fourth Armed service Medical University or college. Dulbecco’s altered Eagle’s medium (DMEM), trypsin, penicillin-streptomycin (penstrep), and fetal bovine serum (FBS) were purchased from GIBCO (Life Technologies, Grand Island, NY). Cell counting kit 8 (CCK-8) assay was purchased from 7sea Biotech (Shanghai, China), Fluorescein Annexin V-FITC/PI double labeling kit was purchased from your Nanjing Jiancheng Bioengineering Institute (Nanjing, China), insulin radioimmunoassay (RIA) kit was purchased from your Institute of Atomic Energy (Beijing, China), and streptozotocin (STZ), LiCl, LY294002, 2,7-dichlorofluorescein diacetate (DCFH-DA), and dihydroethidium (DHE) were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), serum levels Arzoxifene HCl of free fatty acid (FFA), superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione peroxidase (GSH-Px) assay packages were purchased from Jiancheng Bioengineering Institute (Nanjing, China). Palmitate was obtained from Shanghai Macklin Biotechnology (Shanghai, China). Fatty acid-free bovine serum albumin (BSA) was obtained from Shanghai Yeasen Biotechnology (Shanghai, China). Rabbit anti-Nrf2 (#12721, 1?:?1000, Cell Signaling Technology), rabbit anti-IRS-2 (#3089, 1?:?1000, Cell Signaling Technology), rabbit anti-Akt (#9272, 1?:?1000, Cell Signaling Technology), rabbit anti-P-Akt (#4060, 1?:?2000, Cell Signaling Technology), rabbit anti-GSK-3(#12456, 1?:?1000, Cell Signaling Technology), rabbit anti-P-GSK-3(Ser9) (#5558, 1?:?1000, Cell Signaling Technology), rabbit anti-PDX-1 (#5679, 1?:?1000, Cell Signaling Technology), rabbit anti-SOD1 (#2770, 1?:?1000, Cell Signaling Technology), rabbit anti-= 1 : 10), and 25?= 1 : 10). Then, the homogenate was centrifuged at 12000?g for 15?min (at 4C), and the supernatant was isolated and preserved at -80C for the determination of MDA, SOD, and GSH-Px. The pancreatic content of the lipid peroxidation products, MDA, was measured by color changes in thiobarbituric acid reactive substances (TBARS) at 532?nm, and the concentration of TBARS was calculated by referring to the standard curve. SOD activity was assayed at 450?nm on the basis of its ability to inhibit the production of formazan dye resulting from the reaction of WST-1 and superoxide anion. The activity of GSH-Px was measured at 412?nm on the basis of the rate of oxidation of reduced glutathione to oxidized glutathione by H2O2 under the catalysis of GSH-Px. 4. 0.05 was considered to be statistically significant. 5. Results 5.1. Effects of CHS on Glucose Tolerance and Lipid Parameters in Diabetic Rats Physiological parameters were collected to evaluate the changes of the rats in the control and experimental groups. The results show in Physique 1(a) a progressive increase in body weight in the control rats, while the body weight was significantly decreased in the MOD group when compared with the control rats ( 0.05). Treatments Arzoxifene HCl with CHS at the dose of 90?mg/kg and 180?mg/kg significantly increased the body excess weight which was compared with that of MOD group ( 0.05). In MOD rats, the FBG levels (Physique 1(b)) and FIN levels (Physique 1(c)) were increased compared with those of control rats ( 0.05) and decreased by CHS administration significantly ( 0.05). The calculated HOMA-IR index showed a significant increase in the DM Arzoxifene HCl group which compared with the control group ( 0.05). The HOMA-IR index was significantly decreased after CHS treatments (Physique 1(d)). Open in a separate window Physique 1 Effect of CHS on plasma glucose and lipid levels. T2DM model was induced by Slc2a3 HF and STZ; 45, 90, and 180?mg/kg CHS were given through intragastric administration one time every day for 4 consecutive weeks. (a) Body weight, (b) fasting blood glucose (FBG) levels, (c) fasting insulin (FIN) levels, (d) HOMA-IR, (e) glucose tolerance test (OGTT), (f) AUC of OGTT, (g) plasma TC, (h) TG, (i) HDL-c, (j) LDL-c, and (k) FFA were measured accordingly. The columns and errors bars are offered as means SEM. ## 0.01 and # 0.05 compared with the CON group. ?? 0.01 and ? 0.05 compared with the MOD group. To evaluate the metabolic abilities in diabetic rats, OGTT was performed. As the results show in Physique 1(e), the basal plasma glucose level in the HFD-treated group (MOD) was significantly higher than that in Arzoxifene HCl the CON group; after 2?h of glucose administration, the doses of 45, 90, and 180?mg/kg.