Inactivated AMPK-α2 promotes the progression of diabetic brain damage by Cdk5 phosphorylation at Thr485 site.

Changes in brain energy metabolism in diabetes mellitus, including increased insulin resistance and mitochondrial dysfunction, are critically involved in diabetes-related neurodegeneration, and associate with early cognitive impairment as well. The aim of this study is to detect the specific phosphorylated-Thr485- AMP-activated protein kinase (AMPK-α2), regulated by cyclin-dependent kinase 5 (Cdk5) paly the inhibitory functional role of AMPK-α2, Which is maybe the link to the accelerated diabetic brain damage progression. Here, we used GK rats, the type 2 diabetic animal model for in vivo studies and performed In vitro kinase assay, high glucose treatment, -phosphorylated mutation and protein expression in both HEK-293T and HT-22 cell lines. In vitro, the results show that murine wild-type AMPK-α2 was phosphorylated by Cdk5 at a (S/T)PX(K/H/R) phosphorylation consensus sequence, which was associated with decreased AMPK-α2 activity. Surprisingly, mutation of Thr485 to alanine in AMPK-α2 results in the abolished Cdk5 effects, demonstrating that Thr485-phosphorylation is critical to AMPK-α2 inhibition by Cdk5. In addition, these alterations in AMPK-α2-phosphorylation and -activity induced by Cdk5 is specific at Thr485. Furthermore, in GK rats, the increased phosphorylated- Thr 485 of AMPK-α2 results in the decreased AMPK-α2 activity, which is correlated with the apoptosis of neurons in hippocamps. After high glucose treatment, the decreased survival showed in AMPK-α2T485A HT-22 cells compared to AMPK-α2WT. The down-regulated of p-CREB, SNAP25, synaptophysin as well as synapsin-1were shown in both GK rats and HT-22 cell line. Meanwhile, pre-treated with either the specific Cdk5-inhibitor (roscovitine) or the antidiabetic AMPK-α2-inhibitor (metformin) could restore the alterations in neuronal protein expression. Our results suggest that Cdk5-mediated phosphorylated- Thr485 in AMPK-α2 may be involved in the pathogenesis of diabetic brain damage.

Correlation Between Diabetic Cognitive Impairment and Diabetic Retinopathy in Patients With T2DM by 1H-MRS.

Objective: To explore the correlation between diabetic cognitive impairment (DCI) and diabetic retinopathy (DR) through examining the cognitive function and the metabolism of the cerebrum in Type 2 diabetes mellitus (T2DM) by 1H-MRS. Methods: Fifty-three patients with T2DM were enrolled for this study. According to the fundus examination, the patients were divided into the DR group (n = 26) and the T2DM without DR group (T2DM group, n = 27). Thirty healthy adults were selected as a control group (HC group, n = 30). Cognitive function was measured by Montreal Cognitive Assessment (MoCA). The peak areas of N-acetylaspartate (NAA), Cho-line (Cho), Creatine (Cr), and Myo-inositol (mI) as well as their ratios were detected by proton magnetic resonance spectroscopy (1H-MRS). The difference analysis between the three groups was performed by one-way ANOVA. When p < 0.05, LSD-t was applied. A partial correlation analysis (with age as a covariate) was used to analyze the correlation between metabolites in the DR group and MoCA scores. Among all T2DM patients, Chi-square test age, gender, education level, BMI, SBP, DBP, FPG, HbA1c, TC, TG, HDL-C, LDL-C, DR, and DCI correlation were measured. Differences were statistically significant while P < 0.05. Results: 1. The scores of MoCA in the DR group or in the T2DM group were significantly less than those in the HC group (F = 3.54, P < 0.05), and the scores of MoCA in the DR group were significantly less than those in the other groups (F = 3.61, P < 0.05). 2. There were significant differences for NAA in the bilateral hippocampus in DR patients, T2DM patients, and healthy controls (P < 0.05). 3. The NAA/Cr was significantly positively correlated with the score of MoCA in DR patients' left hippocampus (r = 0.781, P < 0.01). 4. Chi-square analysis found that there was a correlation between DR and DCI (x 2 = 4.6, df = 1, p = 0.032, plt: 0.05). There was no correlation between other influencing factors and DCI (P > 0.05). Conclusion: DCI is closely correlated with the DR in patients with T2DM. Hippocampal brain metabolism may have some changes in two sides of NAA in patients with DR, 1H-MRS may provide effective imaging strategies and methods for the early diagnosis of brain damage and quantitative assessment cognitive function in T2DM.