Expression of growth factors and secretory leukocyte protease inhibitor(SLPI) in RAW264.7 cells after lipopolysaccharide(LPS) stimulation
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Secretory leukocyte protease inhibitor (SLPI) was known that bacterial lipopolysaccharide (LPS)-induced product of macrophage. Macrophages play an important role in the development of inflammatory responses by secreting an array of cytokines and chemokines in a tissue microenvironment. To identify the function and relationship between potent growth factors and SLPI after LPS stimulation, we conducted reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of SLPI and growth factors such as VEGF, PDGF, bFGF after 100ng LPS stimulation on the RAW264.7 cells. The result of RT-PCR was showed SLPI mRNA expression was increased from 60 min to 48h in RAW 264.7 cells after incubation with LPS. VEGF and PDGF mRNA was expressed highly at initial stage by LPS stimulation. The expression of bFGF and type I collagen mRNA showed very weakly by LPS treatment. SLPI protein level was increased likely the mRNA levels in lysates by LPS stimulation. Additionally, phase contrast and scanning electron microscopic observation demonstrated that the LPS induce the change of morphology of the RAW264.7 cells. From these results, it suggests that secreted SLPI by LPS treatment may associate with VEGF and PDGF expression in RAW264.7 cells.
Abstract
Secretory leukocyte protease inhibitor (SLPI) was known that bacterial lipopolysaccharide (LPS)-induced product of macrophage. Macrophages play an important role in the development of inflammatory responses by secreting an array of cytokines and chemokines in a tissue microenvironment. To identify the function and relationship between potent growth factors and SLPI after LPS stimulation, we conducted reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of SLPI and growth factors such as VEGF, PDGF, bFGF after 100ng LPS stimulation on the RAW264.7 cells. The result of RT-PCR was showed SLPI mRNA expression was increased from 60 min to 48h in RAW 264.7 cells after incubation with LPS. VEGF and PDGF mRNA was expressed highly at initial stage by LPS stimulation. The expression of bFGF and type I collagen mRNA showed very weakly by LPS treatment. SLPI protein level was increased likely the mRNA levels in lysates by LPS stimulation. Additionally, phase contrast and scanning electron microscopic observation demonstrated that the LPS induce the change of morphology of the RAW264.7 cells. From these results, it suggests that secreted SLPI by LPS treatment may associate with VEGF and PDGF expression in RAW264.7 cells.
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