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Effects of insulin-like growth factor-1 on endoplasmic reticulum stress and autophagy in rat gastric smooth muscle cells cultured at different glucose concentrations in vitro.
Mol Cell Biochem. 2019 Jan; 451(1-2):11-20.MC

Abstract

The purpose of the study was to observe changes in endoplasmic reticulum stress (ERS)- and autophagy-related proteins in gastric smooth muscle tissues of diabetic rats with gastroparesis, investigate the effect of insulin-like growth factor 1 (IGF-1) on ERS and autophagy in rat gastric smooth muscle cells cultured under different glucose concentrations, and explore the influence of IGF-1 on development of diabetic gastroparesis (DGP). After establishing a rat model of DGP, rats were divided into normal control (NC) and 6-week diabetic model (DM6W) groups. Expression of ERS-related and autophagy-related proteins was detected by western blot analysis and immunofluorescence assay in rat gastric smooth muscle tissue and in vitro-cultured rat gastric smooth muscle cells exposed to different glucose concentrations and treatment with IGF-1 for 24 or 48 h. Changes in glucose-regulated-protein-78 (GRP78), growth arrest and DNA damage-inducible gene 153 (CHOP), and microtubule-associated protein 1A/1B light chain 3B (LC3) expression levels were detected by western blot analysis, and GRP78 and LC3 expression were examined by confocal laser-scanning microscopy. In vivo expression levels of GRP78, CHOP, and LC3 were significantly higher in the DM6W group compared with the NC group (p < 0.001). Twenty-four hours after cells were cultured at different glucose concentrations in vitro, expression of GRP78, CHOP, and LC3II/I was significantly higher in the high glucose-treated group compared with the normal glucose group (p < 0.05). After IGF-1 intervention, CHOP and GRP78 expression were significantly higher in the normal glucose + IGF-1 group compared with the normal glucose group (p < 0.01), while no significant difference was found between high glucose and high glucose + IGF-1 groups. LC3II/I expression was significantly lower in the normal glucose + IGF-1 group compared with the normal glucose group, and was significantly lower in the high glucose and high glucose + IGF-1 groups (p < 0.05). After 48 h of culture, CHOP expression was significantly higher and LC3II/I expression was significantly lower in the high glucose group compared with the normal glucose group (p < 0.05), but no significant change in GRP78 expression was observed between these two groups. After IGF-1 intervention, there was no difference in CHOP or GRP78 expression between normal glucose + IGF-1 and normal glucose groups. However, CHOP and GRP78 expression were significantly lower in the high glucose + IGF-1 group compared with the high glucose group (p < 0.05). There was no significant difference in LC3II/I expression between normal glucose + IGF-1 and normal glucose groups, or high glucose + IGF-1 and high glucose groups. Results of confocal laser-scanning microscopy showed significantly lower expression of LC3II/I in the high glucose + IGF-1 group compared with the high glucose group (p < 0.05). ERS and autophagy were involved in the occurrence of DGP. IGF-1 exerted an inhibitory effect on ERS in rat gastric smooth muscle cells cultured under high glucose conditions, and this inhibitory effect increased with time. IGF-1 inhibited the level of autophagy in rat gastric smooth muscle cells cultured under high glucose conditions at early stages, which may be achieved through inhibition of ERS.

Authors+Show Affiliations

Department of Histology and Embryology, Yanbian University College of Medicine, 977 Gongyuan Road, Yanji, 133000, Jilin, China.Department of Histology and Embryology, Yanbian University College of Medicine, 977 Gongyuan Road, Yanji, 133000, Jilin, China.Department of Histology and Embryology, Yanbian University College of Medicine, 977 Gongyuan Road, Yanji, 133000, Jilin, China.Department of Histology and Embryology, Yanbian University College of Medicine, 977 Gongyuan Road, Yanji, 133000, Jilin, China. jinzheng@ybu.edu.cn.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

30008033

Citation

Fang, Xue-Sen, et al. "Effects of Insulin-like Growth Factor-1 On Endoplasmic Reticulum Stress and Autophagy in Rat Gastric Smooth Muscle Cells Cultured at Different Glucose Concentrations in Vitro." Molecular and Cellular Biochemistry, vol. 451, no. 1-2, 2019, pp. 11-20.
Fang XS, Zhang MH, Guo JY, et al. Effects of insulin-like growth factor-1 on endoplasmic reticulum stress and autophagy in rat gastric smooth muscle cells cultured at different glucose concentrations in vitro. Mol Cell Biochem. 2019;451(1-2):11-20.
Fang, X. S., Zhang, M. H., Guo, J. Y., & Jin, Z. (2019). Effects of insulin-like growth factor-1 on endoplasmic reticulum stress and autophagy in rat gastric smooth muscle cells cultured at different glucose concentrations in vitro. Molecular and Cellular Biochemistry, 451(1-2), 11-20. https://doi.org/10.1007/s11010-018-3388-7
Fang XS, et al. Effects of Insulin-like Growth Factor-1 On Endoplasmic Reticulum Stress and Autophagy in Rat Gastric Smooth Muscle Cells Cultured at Different Glucose Concentrations in Vitro. Mol Cell Biochem. 2019;451(1-2):11-20. PubMed PMID: 30008033.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Effects of insulin-like growth factor-1 on endoplasmic reticulum stress and autophagy in rat gastric smooth muscle cells cultured at different glucose concentrations in vitro. AU - Fang,Xue-Sen, AU - Zhang,Mo-Han, AU - Guo,Jun-Yu, AU - Jin,Zheng, Y1 - 2018/07/14/ PY - 2017/12/11/received PY - 2018/06/16/accepted PY - 2018/7/17/pubmed PY - 2019/3/7/medline PY - 2018/7/16/entrez KW - Autophagy KW - CHOP protein KW - Diabetic gastroparesis KW - Endoplasmic reticulum stress KW - GRP78 protein KW - Gastric smooth muscle cells KW - Insulin-like growth factor 1 KW - LC3 protein SP - 11 EP - 20 JF - Molecular and cellular biochemistry JO - Mol Cell Biochem VL - 451 IS - 1-2 N2 - The purpose of the study was to observe changes in endoplasmic reticulum stress (ERS)- and autophagy-related proteins in gastric smooth muscle tissues of diabetic rats with gastroparesis, investigate the effect of insulin-like growth factor 1 (IGF-1) on ERS and autophagy in rat gastric smooth muscle cells cultured under different glucose concentrations, and explore the influence of IGF-1 on development of diabetic gastroparesis (DGP). After establishing a rat model of DGP, rats were divided into normal control (NC) and 6-week diabetic model (DM6W) groups. Expression of ERS-related and autophagy-related proteins was detected by western blot analysis and immunofluorescence assay in rat gastric smooth muscle tissue and in vitro-cultured rat gastric smooth muscle cells exposed to different glucose concentrations and treatment with IGF-1 for 24 or 48 h. Changes in glucose-regulated-protein-78 (GRP78), growth arrest and DNA damage-inducible gene 153 (CHOP), and microtubule-associated protein 1A/1B light chain 3B (LC3) expression levels were detected by western blot analysis, and GRP78 and LC3 expression were examined by confocal laser-scanning microscopy. In vivo expression levels of GRP78, CHOP, and LC3 were significantly higher in the DM6W group compared with the NC group (p < 0.001). Twenty-four hours after cells were cultured at different glucose concentrations in vitro, expression of GRP78, CHOP, and LC3II/I was significantly higher in the high glucose-treated group compared with the normal glucose group (p < 0.05). After IGF-1 intervention, CHOP and GRP78 expression were significantly higher in the normal glucose + IGF-1 group compared with the normal glucose group (p < 0.01), while no significant difference was found between high glucose and high glucose + IGF-1 groups. LC3II/I expression was significantly lower in the normal glucose + IGF-1 group compared with the normal glucose group, and was significantly lower in the high glucose and high glucose + IGF-1 groups (p < 0.05). After 48 h of culture, CHOP expression was significantly higher and LC3II/I expression was significantly lower in the high glucose group compared with the normal glucose group (p < 0.05), but no significant change in GRP78 expression was observed between these two groups. After IGF-1 intervention, there was no difference in CHOP or GRP78 expression between normal glucose + IGF-1 and normal glucose groups. However, CHOP and GRP78 expression were significantly lower in the high glucose + IGF-1 group compared with the high glucose group (p < 0.05). There was no significant difference in LC3II/I expression between normal glucose + IGF-1 and normal glucose groups, or high glucose + IGF-1 and high glucose groups. Results of confocal laser-scanning microscopy showed significantly lower expression of LC3II/I in the high glucose + IGF-1 group compared with the high glucose group (p < 0.05). ERS and autophagy were involved in the occurrence of DGP. IGF-1 exerted an inhibitory effect on ERS in rat gastric smooth muscle cells cultured under high glucose conditions, and this inhibitory effect increased with time. IGF-1 inhibited the level of autophagy in rat gastric smooth muscle cells cultured under high glucose conditions at early stages, which may be achieved through inhibition of ERS. SN - 1573-4919 UR - https://www.unboundmedicine.com/medline/citation/30008033/Effects_of_insulin_like_growth_factor_1_on_endoplasmic_reticulum_stress_and_autophagy_in_rat_gastric_smooth_muscle_cells_cultured_at_different_glucose_concentrations_in_vitro_ L2 - https://doi.org/10.1007/s11010-018-3388-7 DB - PRIME DP - Unbound Medicine ER -