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| The effect of high glucose on autophagy of cultured murine myotubes |
| LUO Shengnan1, SU Zhen2, HUANG Peipei3, CHEN Ruyi4, ZUO Yidan5, HU Feifei6 |
| Department of Nephrology, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325015, China. |
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| Cite this article: |
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LUO Shengnan,SU Zhen,HUANG Peipei, et al. The effect of high glucose on autophagy of cultured murine myotubes[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2019, 49(5): 313-320.
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Abstract Objective: To observe the effect of high glucose on the expression of autophagy in myotubes, and to investigate the expression of autophagy in myotubes cultured with high glucose for different time periods. Methods: Murine C2C12 myoblasts were induced into myotubes using differentiation medium, and myotubes were cultured with normal (5.5 mmol/L) and high dose (17 mmol/L or 30 mmol/L) of glucose, and cells were collected at 6, 12, 24, 36, 48, 60, and 72 h to detect autophagy-related proteins and mRNA expression. Additionally, murine myoblasts were transfected with tandem fluorescently labeled LC3 (mRFP-GFP-LC3) lentiviral vector. The red and green fluorescent spots in the cells were observed by laser confocal microscopy to monitor autophagic flux. Results: The autophagic flux in myotubes exposed to high glucose was divided into two phases. When myotubes incubated for 24 h with high glucose, the protein expression of LC3 and Beclin-1 was lower than that of the normal group (P<0.05); When C2C12 myotubes were cultured with 30 mmol/L of glucose for 48, 60 and 72 h, respectively, the levels of LC3 and Beclin-1 mRNA were significantly increased (P<0.05). In addition, localization and quantitative analysis of GFP and RFP fluorescent puncta in myotubes also demonstrated a significant increase in the synthesis of autophagosomes and autolysosomes after 48, 60 and 72 h of 30 mmol/L glucose culture, respectively. Conclusion: High glucose plays an important effect on autophagy in myotubes. Long-term hyperglycemia may eventually induce excessive autophagy.
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Received: 20 December 2018
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