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2018 ; 17
(4
): 5642-5651
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High glucose induces podocyte epithelial?to?mesenchymal transition by
demethylation?mediated enhancement of MMP9 expression
#MMPMID29436620
Ling L
; Chen L
; Zhang C
; Gui S
; Zhao H
; Li Z
Mol Med Rep
2018[Apr]; 17
(4
): 5642-5651
PMID29436620
show ga
Abnormal expression of matrix metalloproteinase 9 (MMP9) is correlated with
podocyte epithelial-to---mesenchymal transition (EMT) in diabetic nephropathy
(DN). However, the mechanisms underlying this process are not well defined.
Site?specific demethylation may sustain high expression levels of target genes.
In the present study, in order to investigate the association between DNA
demethylation of MMP9 promoter and podocyte EMT in DN, human podocytes were
cultured in high?glucose (HG) medium and a rat model of DN was established by
intraperitoneal injection of streptozotocin (STZ) to determine whether
site?specific demethylation of the MMP9 promoter was involved in regulating
podocyte EMT in DN. The MTT assay was used to assess the effects of HG culture on
the growth of podocytes, and the demethylation status of the MMP9 promoter was
assessed by bisulfite sequencing polymerase chain reaction. mRNA and protein
expression levels of MMP9, ??smooth muscle actin (??SMA), podocalyxin and
fibronectin?1 in podocytes were assessed by reverse transcription?quantitative
PCR (RT?qPCR) and western blot analyses. The results demonstrated that HG
treatment up regulated the expression of MMP9, ??SMA and fibronectin?1, but down
regulated the expression of podocalyxin in podocytes. The MMP9 promoter region
was revealed to contain a variety of demethylated CpG sites, and HG treatment
reduced the rate of MMP9 promotermethylation, which, in turn, enhanced its
promoter activity. In summary, these data suggested that demethylation of the
MMP9 promoter may serve an important role in podocyte EMT in DN. The
demethylation status of the MMP9 promoter maybe used as an important prognostic
marker of DN in clinic.
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