Retracted: Bilobalide alleviates tumor necrosis factor‐alpha‐induced pancreatic beta‐cell MIN6 apoptosis and dysfunction through upregulation of miR‐153

Abstract

Type 1 diabetes mellitus (T1DM) is a systemic disease and one classical type of total DM. Bilobalide (BB) is constituted of EGb 761. Our purpose was identifying the role of BB in TIDM in the current study. MIN6 cells were treated by TNF-α; then, viability, apoptosis, and insulin secretion were assessed by performing Cell Counting Kit-8 assay, flow cytometry, glucose-stimulated insulin secretion assay, and western blot.The effects of BB were assessed to identify its function. Further, the above mentioned parameters were reassessed when silencing miR-153. TNF-α declined viability and insulin secretion as well as raised apoptosis and inducible nitric oxide synthase (iNOS) expression in MIN6 cells. BB alleviated the apoptosis and dysfunction induced by TNF-α. MiR-153 expression was elevated by BB when induced by TNF-α. Increase of viability and insulin secretion as well as decline of apoptosis and iNOS induced by BB treatment was alleviated by silencing miR-153. The rates of p/t-p70S6K, p/tmammalian target of rapamycin (mTOR) and p/t-adenosine monophosphateactivated protein kinase (AMPK) were raised by BB and suppressed by silencing miR-153 under TNF-α induced condition. BB raised viability and insulin secretion, declined apoptosis and iNOS expression by up-regulating miR-153. Furthermore, BB activated AMPK/mTOR pathway by up-regulating miR-153.

Suppressive role of E3 ubiquitin ligase FBW7 in type I diabetes in non-obese diabetic mice through mediation of ubiquitination of EZH2

“…To determine the cytokine levels, splenocytes were cultured in DMEM supplemented with 10% FBS (0.2 mL) at a final concentration of 1.5 × 10 6 cells/well. The supernatant was collected after 48 h and cytokines, IL-17A (M1700), IL-2 (M2000), IL-4 (M4000B), and IFN-γ (MIF00), were assessed using a quantitative ELISA kit (R&D Systems, Inc., Minneapolis, MN, USA) [43].…”

“…First, the cell suspension was placed in a 96-well plate with a concentration of 5 × 10 4 cells/well, and pre-warmed in the incubator (37°C, 5% CO 2 ). Next, CCK-8 solution (10 μL) was added to each well and the plate was incubated for 1-4 h. Finally, the absorbance at 450 nm was recorded on a microplate reader (Bio-Rad, Hercules, CA, USA) [43].…”

E3 ubiquitin ligase FBW7-mediated ubiquitination of EZH2 prevents type I diabetes in NOD mice

“…To determine the cytokine levels, splenocytes were cultured in DMEM supplemented with 10% FBS (0.2 mL) at a final concentration of 1.5 × 10 6 cells/ well. The supernatant was collected after 48 hrs and cytokines were assessed using quantitative ELISA kit (BD Bioscience, San Jose, CA, USA) [18].…”

“…GAPDH (ab9485, 1:500) was served as the internal reference. Each experiment was conducted in triplicate [18].…”

“…Next, CCK-8 (10 µL) was added to each well and the plate was incubated for 1-4 hrs. Finally, the absorbance at 450 nm was recorded on a microplate reader (Bio-Rad, Hercules, CA, USA) [18].…”