Di-san junyi daxue xuebao (May 2021)

Effect of oncostatin M on proliferation and secretion of extracellular matrix in human dermal fibroblasts in high glucose condition

  • LIU Lulu,
  • FU Xueming,
  • XUE Biyu,
  • XUE Bin

DOI
https://doi.org/10.16016/j.1000-5404.202011253
Journal volume & issue
Vol. 43, no. 10
pp. 955 – 963

Abstract

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Objective To investigate the effects of oncostatin M (OSM) on the proliferation and extracellular matrix secretion of human dermal fibroblasts (HDFs) in high glucose environment and its possible mechanism. Methods ① Establishment of hyperglycemia model: HDFs were treated with 5, 10, 15, 20 and 25 mmol/L glucose, respectively. MTT assay was used to detect the proliferation activity of cells cultured in high glucose for 24, 36 or 48 h. Western blotting was used to detect the protein levels of ERK, p-ERK and CyclinD1 in the HDFs treated with 5 or 20 mmol/L glucose for 24, 36 and 48 h. ② Optimal concentration of OSM on hyperglycemia model: the cells were treated with 20 mmol/L glucose (control group), and 20 mmol/L glucose+25, 50, 100, 200 ng/mL OSM, respectively. MTT assay was used to detect the proliferation of above cells cultured for 24, 36 and 48 h. The protein expression of CollagenⅠ, CollagenⅢ and fibronectin was detected by Western blotting. ③ Mechanism of OSM on hyperglycemia model: the cells were divided into control group, high glucose group, high glucose+OSM group, and high glucose+OSM+ inhibitor PD98059 group. Cell cycle was determined by flow cytometry. The protein levels of ERK, p-ERK, CyclinD1, CollagenⅠ, CollagenⅢ, fibronectin and other related factors were detected by Western blotting. q-PCR was used to measure the levels of ERK1/2, CyclinD1, CollagenⅠ, CollagenⅢ and fibronectin in each group. Results Treatment of 20 mmol/L glucose resulted in significantly decreased proliferation activity (P < 0.001), increased proportion of G1 cells (P < 0.01) and decreased proportion of S-phase cells (P < 0.01), and reduced expression of ERK, p-ERK, CyclinD1, CollagenⅠ, CollagenⅢ and fibronectin (P < 0.01, P < 0.05) when compared with the control cells. While treatment of 100 ng/mL OSM for 48 h enhanced the proliferation activity (P < 0.001), decreased the proportion of G1 cells (P < 0.05) but elevated the proportion of S-phase cells (P < 0.05), and raised protein levels of ERK, p-ERK, CyclinD1, CollagenⅠ, CollagenⅢ and fibronectin cells (all P < 0.01). q-PCR indicated similar results as Western blotting, and statistical differences were seen in the results of mRNA levels (all P < 0.05). When ERK pathway specific small molecule inhibitor PD98059 blocked ERK signaling pathway, the above effects were significantly reduced (P < 0.05). Conclusion OSM may reverse the inhibitory effect of high glucose on the proliferation and extracellular matrix secretion through ERK signaling pathway in HDFs.

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