[Role of Akt/NF-kappa B signal transduction pathway in murine glomerular mesangial cell proliferation induced by immune complex].
OBJECTIVE
To explore whether immune complex (IC) can directly induce glomerular mesangial cells(MCs) proliferation and the role of Akt/NF-kappa B signal pathway in the proliferation.


METHODS
The mice were divided into control, stimulation and oligodeoxynucleotide(ODN) groups. In ODN group, MCs isolated from mice were transfected with Akt1 sense, mismatched or antisense ODN for 8 h, respectively, by using lipofectin, control and stimulation groups were incubated with lipofectin for 8 h. Then stimulation and ODN groups were incubated with aggregated IgG(AIgG)(a standard IC model), while the control group with monomeric IgG. MTT colorimetry was used to detect MCs proliferation. Distribution of MCs in cell cycle was analyzed by flow cytometry. Cyclin D1 mRNA and its protein expression were determined by RT-PCR and Western blot, respectively.The activity of NF-kappa B in MCs was determined by EMSA.


RESULTS
AIgG activated NF-kappa B, upregulated cyclin D1 mRNA and its protein expression, and induced majority of MCs to enter S-phase in cell cycle. Akt1 antisense ODN specifically decreased AIgG-induced NF-kappa B activation, cyclin D1 mRNA and its protein expression, and then inhibited MCs to progress to S-phase and cell proliferation. Sense ODN and mismatched ODN had no such effects.


CONCLUSIONS
IC can directly stimulate MCs proliferation through Akt/NF-kappa B signal pathway, suggesting that NF-kappa B probably be a useful molecule for targeted therapy in IC-mediated MC overproliferation.
Wang
Bi-Fei
BF
Department of Nephrology, First Affiliated Hospital, Sun Yat-sen University, Key Clinical Laboratory for Renal Diseases, Ministry of Education, Guangzhou 510080, China.
Xu
Han-Shi
HS
Li
You-Ji
YJ
Ye
Ren-Gao
RG
Kong
Qing-Yu
QY
Yu
Xue-Qing
XQ
D000818:Animals; D000936:Antigen-Antibody Complex / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D019938:Cyclin D1 / Q000096:biosynthesis* / Q000235:genetics; D005920:Glomerular Mesangium / Q000166:cytology / Q000378:metabolism*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D016328:NF-kappa B / Q000378:metabolism*; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology*; D017346:Protein Serine-Threonine Kinases / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology*; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology*; D051057:Proto-Oncogene Proteins c-akt; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D016196:S Phase; D015398:Signal Transduction; D014162:Transfection; D015854:Up-Regulation
D000936:Antigen-Antibody Complex; D016328:NF-kappa B; D020319:Oligodeoxyribonucleotides, Antisense; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D019938:Cyclin D1; D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt