Role of acid sphingomyelinase in silica-induced pulmonary fibrosis model in vitro

Feng Jin, Sheng Wei Jiang, Ming Zeng, Xing Xuan He, Edward H. Schuchman, An Wang, Lei Luo, Fu Wang, Fang Xiao, Lan Guan, Xin Min Liu, Cai Gao Zhong

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5 Scopus citations

Abstract

OBJECTIVE: To observe the level and change rule of acid sphingomyelinase (ASM) in silica-induced fibrosis in mouse embryonic fibroblast (NIH3T3), and to explore the functional role of ASM in silica-induced pulmonary fibrosis in vitro. METHODS: Pulmonary alveolar macrophages (PAM) were treated with SiO 2 200 mg·L-1 for 12 h, and the medium supernatant was collected to stimulate NIH3T3 for 6, 12, 18, 24, 36, and 48 h. ELISA assay was applied to determine the contents of interleukin-8 (IL-8), tumor necrosis factor (TNF-α) and transforming growth factor-β1 (TGF-β1) in the supernatant collected from the culture medium of PAM. ASM activity of NIH3T3 was assayed using high-performance liquid chromatographic (HPLC). The protein expression level of type III collagen was determined by western blotting. RESULTS: The levels of IL-8, TNF-α and TGF-β1 in PAM exposed to SiO2200 mg·L-1 were obviously increased (P <0.01). ASM activity of NIH3T3 was increased at the time point of 24, 36 and 48 h after stimulated with the supernantant(P<0.05), and the peak was appeared at 36 h. In the group treated with desipramine 1.25 μmol·L-1, ASM activity was decreased with the prolonged time. Western blotting results revealed that the expression of type III collagen was up-regulated at the time point of 24, 36 and 48 h after stimulation (P<0.05), and the peak appeared at 36 h. And the expression of type III collagen was down-regulated with the prolonged time in the group treated with desipramine 1.25 μmol·L-1. CONCLUSION: Inhibition of ASM activity may have a certain effect on improving pulmonary fibrosis.

Original languageEnglish
Pages (from-to)205-209
Number of pages5
JournalChinese Journal of Pharmacology and Toxicology
Volume28
Issue number2
DOIs
StatePublished - Apr 2014

Keywords

  • Acid sphingomyelinase
  • Macrophage
  • Pulmonary fibrosis
  • Silicon
  • Type III collagen

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