Key genes and proteins involved in CTCM-reducing microvascular endothelial cell permeability induced by SLT-IIv using gene chips and DIGE.
An Affymetrix mouse genome array and differential in-gel electrophoresis (DIGE) techniques were used to investigate the pharmacological mechanisms of a mixture of herbs, designated CTCM, a compound of traditional Chinese medicine, for the treatment of increased permeability in mouse intestinal microvascular endothelial cells (MIMECs) induced by the Shiga-like toxin type II variant (SLT-IIv). MIMECs were challenged with 10microg/ml SLT-IIv for 12h and then treated with CTCM at a concentration of 200microg/ml for 12h. Total RNA and proteins from each treatment group were extracted from cultured MIMECs for analysis by the Affymetrix GeneChip Mouse Genome 430 2.0 microarray and DIGE. The results obtained demonstrated that there were one genes downregulated and one genes upregulated, one protein downregulated and four proteins upregulated in the SLT-IIv group compared to the control group. In the CTCM group, four genes were upregulated, three genes were downregulated, a single protein was downregulated and a single protein was upregulated when compared to the control group. When the CTCM-treated group was compared to the SLT-IIv group, expression of one gene was found to be increased, and all other genes were decreased, with five proteins downregulated. Analysis of the data suggested that CTCM specifically and effectively reduced microvascular endothelial cell permeability to SLT-IIv in the treatment of pig edema disease. In the CTCM-treated group, hspa9 expression was increased in both gene chip and DIGE analysis, so it may be a key protein in reducing cell permeability and utilized in medical treatments.
Yi
Pengfei
P
College of Animal Science and Veterinary Medicine, Jilin University, Changchun, Jilin 130062, PR China. yipengfei1982@yahoo.com.cn
Guo
Yang
Y
Wang
Xin
X
Mu
Xiang
X
Wei
Xubin
X
D000818:Animals; D002199:Capillary Permeability / Q000187:drug effects* / Q000276:immunology; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D015180:Electrophoresis, Gel, Two-Dimensional; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D004927:Escherichia coli Infections / Q000188:drug therapy / Q000276:immunology / Q000382:microbiology / Q000662:veterinary*; D005767:Gastrointestinal Diseases / Q000188:drug therapy / Q000276:immunology / Q000382:microbiology / Q000662:veterinary*; D005786:Gene Expression Regulation / Q000187:drug effects; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D012313:RNA / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D022641:Shiga Toxin 2 / Q000235:genetics / Q000633:toxicity*; D013552:Swine; D013553:Swine Diseases / Q000188:drug therapy / Q000235:genetics* / Q000276:immunology / Q000382:microbiology
D004365:Drugs, Chinese Herbal; D022641:Shiga Toxin 2; C405162:Shiga-Like Toxin IIv; D012313:RNA