[Effect of tissue factor pathway inhibitor on apoptosis of rat mesangial cells and Fas and bcl-2 expression].
OBJECTIVE
To study the biological impact and mechanism of recombinant tissue factor pathway inhibitor (rTFPI) on apoptosis of rat kidney mesangial cells (MsC).


METHODS
TFPI expression in human glomerular minor lesion (GML), mesangial proliferative glomerulonephritis (MPGN) and cultured rat MsC was detected using immunohistochemistry and immunofluorescence, respectively. Rat MsC were incubated with rTFPI and its variant peptides. Morphological changes of apoptosis were investigated by Hoechst 33258 and the apoptotic rate was assessed by flow cytometry. DNA fragmentation and effect of rTFPI on expression of caspase-3, Fas and bcl-2 were studied using gel electrophoresis and Western blot respectively.


RESULTS
The expression of TFPI in MPGN was higher than that in GML. TFPI was expressed in cultured rat mesangial cells. Apoptosis of MsC was induced by rTFPI, especially by its C-termianl, in a dose- and time-dependent manner. Apoptosis ratios of MsC treated with rTFPI were 2.1, 3.0 and 4.9 times more than control, respectively. Expression of gene caspase-3 and Fas was up-regulated in a dose-dependent manner wherease bcl-2 expression did not show any changes.


CONCLUSIONS
rTFPI induces apoptosis in cultured rat mesangial cells by its C-terminal possibly via Fas/FasL pathway.
Lin
Yi-feng
YF
Department of Pathology, Shanghai Medical College, Fudan University, Shanghai 200032, China.
Ma
Duan
D
Liang
Wang
W
Jiang
Tao
T
Zhao
Zhong-hua
ZH
Zhang
Nong
N
D000818:Animals; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D053148:Caspase 3 / Q000378:metabolism*; D002478:Cells, Cultured / Q000187:drug effects; D053938:DNA Fragmentation; D005434:Flow Cytometry; D006801:Humans; D008074:Lipoproteins / Q000378:metabolism / Q000494:pharmacology*; D050527:Mesangial Cells / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D010455:Peptides / Q000494:pharmacology; D051381:Rats; D051025:bcl-Associated Death Protein / Q000378:metabolism* / Q000494:pharmacology
C494821:Bad protein, rat; D008074:Lipoproteins; D010455:Peptides; D051025:bcl-Associated Death Protein; C051928:lipoprotein-associated coagulation inhibitor; D053148:Caspase 3