Effects of secreted protein acidic and rich in cysteine peptide on change of ultramicrostructure and extracellular matrix secretion of human mesangial cells cultured in vitro
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    Abstract:

    Objective: To investigate the effects of secreted protein acidic and rich in cysteine (SPARC) peptide on ultramicrostructure and extracellular matrix secretion of human mesangial cells cultured in vitro. Methods: Human mesangial cells (HMC) were incubated at the presence of SPARC peptide (50μg/ml) for 48 h and 96 h separately; HMC cultured without SPARC peptide was taken as control. The cell ultramicrostructure was observed by transmission electron microscope. Real-time fluorescent quantitative PCR (RT-PCR) was used to detect mRNA levels of collagen typeⅠ (ColⅠ ), collagen type Ⅳ (ColⅣ), fibronectin (FN), and laminin (LN) 96 hours after culture, and the results were compared between the 2 groups. The secreted levels of ColⅠ , ColⅣ, FN, and LN protein were measured and compared by enzyme-linked immunosorbent assay (ELISA) in the 2 groups. Results: There was no obvious change in HMC in the control group. After cultured for 48 hours, a few HMC in experimental group showed initial appearance of apoptosis; after cultured for 96 h, a great number of HMC had a typical apoptotic appearance and there were typical apoptotic bodies under electron microscope. RT-PCR showed that, compared with the control group, the experimental group had significantly decreased levels of ColⅠ , ColⅣ and FN mRNA (P〈0.05, P〈0. 01) ; ELISA results showed that the secretions of ColⅠ , ColⅣ, and FN protein in the experimental group were greatly lower than those in the control garoup (P〈0.05, P〈0.01). Conclusion: SPARC peptide can effectively induce apoptosis of human mesangial cells and down regulate the secretion of extracellular matrix in vitro

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History
  • Received:October 19,2006
  • Revised:December 18,2006
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  • Online: January 20,2007
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