[Effect of nuclear factor-kappaB on signal transduction of passively sensitized human airway smooth muscle cells proliferation].
OBJECTIVE
To investigate whether nuclear factor-kappaB (NF-kappaB) contributes to passively sensitized human airway smooth muscle cell (HASMCs) proliferation, and whether it is the downstream factor of activated protein kinase C (PKC).


METHODS
HASMCs in culture were passively sensitized with 10% serum from asthmatic patients, with non-asthmatic human serum treated HASMCs as the control. NF-kappaB specific inhibitor pyrrolidine dithiocarbamate (PDTC) and PKC agonist phorbol 12-myristate 13-acetate (PMA) were used to intervene HASMCs exposed to asthmatic serum and non-asthmatic control serum. The proliferation of HASMCs was examined by cell cycle analysis, MTT colorimetric assay and proliferating cell nuclear antigen (PCNA) immunofluorescence staining respectively. NF-kappaB activity was detected by NF-kappaBp65 immunofluorescence staining and electrophoretic mobility shift assay (EMSA) respectively.


RESULTS
(1) The percentage of S phase, A value, the positive expression rate of PCNA, the positive expression rate of NF-kappaBp65 and EMSA value in HASMCs passively sensitized with asthmatic serum were (21.78 +/- 2.79)%, 0.466 +/- 0.058, (67.5 +/- 8.5)%, (12.6 +/- 2.2)% and 32 781 +/- 9499 respectively. They were significantly increased compared with those of the control serum group (P < 0.05). After previously treated with PDTC, the above figures were decreased to (16.37 +/- 3.05)%, 0.389 +/- 0.035, (53.4 +/- 5.1)%, (4.9 +/- 1.3)% and 3934 +/- 937 respectively (P < 0.05). (2) After HASMCs were treated with both PMA and asthmatic serum, the percentage of S phase, A value, the positive expression rate of PCNA, the positive expression rate of NF-kappaBp65 and EMSA value were (25.52 +/- 3.38)%, 0.572 +/- 0.054, (81.2 +/- 10.2)%, (26.5 +/- 5.0)% and 71 654 +/- 12 293 respectively. After previously treated with PDTC, the above figures were (16.42 +/- 2.72)%, 0.386 +/- 0.031, (54.2 +/- 5.3)%, (5.9 +/- 1.4)% and 4808 +/- 1084 respectively. The difference was significant (P < 0.05).


CONCLUSIONS
NF-kappaB may contribute to the proliferation of HASMCs passively sensitized with human asthmatic serum, which involves the PKC/NF-kappaB signal pathway.
Xu
Shu-yun
SY
Department of Respiratory Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Xu
Yong-jian
YJ
Zhang
Zhen-xiang
ZX
Ni
Wang
W
Chen
Shi-xin
SX
D000328:Adult; D001249:Asthma / Q000473:pathology*; D049109:Cell Proliferation; D002478:Cells, Cultured; D005260:Female; D006801:Humans; D007116:Immunization, Passive; D008297:Male; D008875:Middle Aged; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism / Q000502:physiology*; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D011493:Protein Kinase C / Q000502:physiology; D015398:Signal Transduction / Q000502:physiology*
D016328:NF-kappa B; D018809:Proliferating Cell Nuclear Antigen; D011493:Protein Kinase C