Effects Of EP On The Expression Of MMP-9、Col Ⅳin Cultured Renal Mesangial Cells

Objective: To observe the effects of Ethyl pyruvate (EP) to the expression of matrixmetalloproteinase-9(MMP-9) and type Ⅳ collagen (Col Ⅳ) in rat mesangialcell line (HBZY-1). And investigate the possible mechanism.Methods: HBZY-1were incubated with medium containing different concentration ofglucose in vitro for24hours or with high-glucose20mmol/L for differenttimes, to detect the expression of MMP-9、 Col ⅣmRNA byReactive-transcription polymerase chain reaction (RT-PCR); Afterpre-treated with different doses of EP、PDTC (the specific inhibitor ofNF-κB) and different times of EP, the expression of MMP-9、Col ⅣmRNAand proteins were measured by RT-PCR and western-blotting;Immunofluorescence assay was used to estimate the expression of NF-κBp65.Results:(1)When the glucose density between5.6～30mM or the the20mM highglucose at different times, the mRNA expression of MMP-9wasdown-regulated (P<0.05)，Col Ⅳ mRNA was up-regulated (P<0.05). Both ofthem were in a dose-and time-dependent manner.(2) The high-glucoseinduced down-regulation of MMP-9and up-regulation Col Ⅳwere largelyblocked by EP and PDTC with concentration dependent. The expression ofMMP-9、Col Ⅳbetween EP5mmol/L and10mmol/L was similar, suggestedthat5mmol/L may be the appropriate concentration.(3) The signal of NF-κBwas translocated from the cytoplasm to the nuclei induced by high glucose,EP and PDTC restrained the activation of NF-κB and blocked the nucleartranslocation of NF-κB obviously. Conclusions:(1) The expression of MMP-9、Col Ⅳregulated by high glucose conditionin mesangial cells, may contribute to the development and progression ofdiabetic nephropathy.(2) EP、PDTC could regulate the expression ofMMP-9and Col Ⅳ in some dose-and-time dependent manner, suggestedthat EP and PDTC against diabetic nephropathy may be partially through itsanti-inflammatory and anti-fibrosis effect.(3) The role of EP and PDTCabove may be through the inhibition of NF-κB signal pathway.