Insulin inhibits the JNK mediated cell death via upregulation of AKT expression in Schwann cells grown in hyperglycemia

doi:10.1007/s11515-018-1492-4

Front. Biol.

2018, Vol. 13

Issue (2) : 137-144 https://doi.org/10.1007/s11515-018-1492-4

RESEARCH ARTICLE

Insulin inhibits the JNK mediated cell death via upregulation of AKT expression in Schwann cells grown in hyperglycemia

Mallahalli S. Manu, Kuruvanthe S. Rachana, Gopal M. Advirao(

)

Department of Biochemistry, Davangere University, Davangere, Karnataka, India

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Abstract

BACKGROUND: Schwann cells (SCs) are the glial cells of the peripheral nervous system, which forms a thick insulating structure around the axons. Hyperglycemia is known physiologic conditions in both type I and type II diabetes which causes diabetic neuropathy. But the SC possesses insulin receptors even though glucose uptake is independent of insulin. Since the insulin level is highly altered in diabetes, it is of greater importance to evaluate their role in the Schwann cell survival and death.

METHODS: Schwann cells were isolated from neonatal pups and grown with and without insulin in hyperglycemic medium to mimic diabetic condition for 24 and 48 h. We studied the cell viability using 3 (4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT) and mitochondrial membrane potential (MMP) assay at different time interval on SCs. We also studied the protein and gene expression of Protein Kinase B (AKT) and Jun N-terminal kinase (JNK), which are greatly involved in cell survival and cell death respectively.

RESULTS: The result shows that, high glucose levels for 48 h decrease the SC viability. Hyperglycemic condition induces the SC death by increasing the JNK expression which in turn reduces the MMP of glial cells. However, insulin administration for SCs grown in high glucose condition can reduce the JNK expression by activating AKT signaling pathway.

CONCLUSION: These observations demonstrate that the proper insulin balance is required for Schwann cells survival in hyperglycemic condition. Therefore, altered insulin signaling can be one of the reasons for demyelination of peripheral neurons in diabetic neuropathy.

Keywords insulin schwann cells apoptosis JNK AKT diabetic peripheral neuropathy

Corresponding Author(s): Gopal M. Advirao

Online First Date: 09 May 2018 Issue Date: 28 May 2018

Cite this article:

Mallahalli S. Manu,Kuruvanthe S. Rachana,Gopal M. Advirao. Insulin inhibits the JNK mediated cell death via upregulation of AKT expression in Schwann cells grown in hyperglycemia[J]. Front. Biol., 2018, 13(2): 137-144.

URL:

https://academic.hep.com.cn/fib/EN/10.1007/s11515-018-1492-4
https://academic.hep.com.cn/fib/EN/Y2018/V13/I2/137

Fig.1 (A and B) Viability of SCs in presence and absence of 10nM of insulin at different glucose concentration for 24 h and 48 h were determined by MTT assay.

Fig.2 Effect of high glucose in presence and absence of insulin on mitochondrial membrane potential of SCs at different time intervals was measured using 10 mM JC-1 ﬂuorescence dye. All the experiments were done in triplicates.

Fig.3 SCs were grown in normal condition (C), high glucose conditions (H-G) and high glucose with 10nM of insulin (H-G+ I) for 24 h and 48 h. (A) Western blot analysis showing relative fold change in pJNK proteins expression. (B) Fold changes were presented as bar chart. (C) Real time studies showing relative mRNA expression of JNK. RPL19 is used as internal control. * indicates p<0.05.

Fig.4 Immunofluorescence expression of pJNK protein in SCs grown for 24 h and 48 h in normal medium [(control) A and B], high glucose medium [(40 mM glucose) C and D] and insulin added high glucose medium [(40 mM glucose+ 10 nM insulin) E and F]. Scale bar: 50 µm.

Fig.5 SCs were grown in normal condition (C), high glucose conditions (H-G) and high glucose with 10nM of insulin (H-G+ I) for 24 h and 48 h. (A) Western blot analysis showing relative fold change in pAKT proteins expression. (B) Fold changes were presented as bar chart. (C) Real time studies showing relative mRNA expression of AKT. RPL19 is used as internal control. * indicates p<0.05. Scale bar: 50 µm.

Fig.6 Immunofluorescence expression of pAKT protein in SCs grown for 24 h and 48 h in normal medium [(control) A and B], high glucose medium [(40 mM glucose) C and D] and insulin added high glucose medium [(40 mM glucose+ 10 nM insulin) E and F]. Scale bar: 50 µm.

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