[Study on effect of anisodamine on expressions of tissue factor and plasminogen activator-1 inhibitor in vascular endothelial cells and its mechanisms].
OBJECTIVE
To explore the mechanism of anisodamine in treating infectious shock through studying effect of anisodamine on endotoxin lipopolysaccharide (LPS) induced expression of tissue factor (TF) and plasminogen activator inhibitor type 1 (PAI-1) in vascular endothelial cells (EC).


METHODS
Human umbilical vein endothelial cells (HUVEC) were cultured by trypsin digestion method. PAI-1 was measured in the conditioned medium of HUVEC by a specific enzyme-linked immunosorbent assay (ELISA), whereas TF activity was measured in the lysates of these cells by using a single step clotting assay. Specific mRNA expressions were determined by Northern blotting. In order to evaluate a possible contribution of the nuclear factor-kappa B (NF-kappa B) pathway on the transductive effects observed, electrophoretic mobility shift assays (EMSA) were performed using nuclear extracts from HUVEC and NF-kappa B binding oligonucleotides.


RESULTS
LPS could significantly strengthen the expression of HUVEC PAI-1 protein and TF activity and its mRNA, this effect of LPS could be markedly weakened after adding Anisodamine dose-dependently. Anisodamine could also completely block the LPS induced NF-kappa B DNA binding activity in nuclear extracts from HUVEC.


CONCLUSIONS
The possible mechanism of anisodamine in treating infectious shock may be through antagonizing LPS induced HUVEC TF and PAI-1 expression, and the antagonism might be, at least partially, transduced by path of NF-kappa B.
Ruan
Qiu-rong
QR
Department of Pathology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030. ruanqiurong@sina.com
Song
Jian-xin
JX
Deng
Zhong-duan
ZD
D002478:Cells, Cultured; D017077:Culture Media, Conditioned; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D016328:NF-kappa B / Q000378:metabolism; D017395:Plasminogen Activator Inhibitor 1 / Q000096:biosynthesis* / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D012991:Solanaceous Alkaloids / Q000494:pharmacology*; D013925:Thromboplastin / Q000096:biosynthesis* / Q000235:genetics; D014471:Umbilical Veins / Q000166:cytology
D017077:Culture Media, Conditioned; D004365:Drugs, Chinese Herbal; D016328:NF-kappa B; D017395:Plasminogen Activator Inhibitor 1; D012333:RNA, Messenger; D012991:Solanaceous Alkaloids; C003922:anisodamine; D013925:Thromboplastin