Triptolide suppresses IL-1beta-induced chemokine and stromelysin-1 gene expression in human colonic subepithelial myofibroblasts.
OBJECTIVE
To examine the inhibitive effects of triptolide on the expression of IL-8, monocyte chemotactic protein (MCP)-1, and matrix metalloproteinases (MMP)-3 in subepithelial myofibroblasts (SEMF) stimulated with IL-1beta.


METHODS
SEMF cultures were established from normal colons in patients who underwent gut resection for colorectal carcinoma. Chemokine and MMP-3 expressions were determined by ELISA and RT-PCR. The cytosolic amount of phosphorylation of I kappa B-alpha(p-I kappa B-alpha) was determined by Western blotting. The DNA binding capacity of NF-kappa B was evaluated by electrophoretic mobility shift assays.


RESULTS
IL-1beta stimulated protein and mRNA expression of IL-8, MCP-1, and MMP-3 in SEMF. Triptolide inhibited these effects of IL-1beta in a dose-dependent manner. Mechanistic studies revealed that triptolide markedly decreased IL-1beta -induced NF-kappa B DNA binding capacity and cytosolic amount of p-I kappa B-alpha. These results showed that triptolide inhibited IL-1beta -induced chemokine and MMP-3 expression in SEMF through the NF-kappa B pathway.


CONCLUSIONS
Triptolide inhibited IL-1beta -induced chemokine and MMP-3 expression in SEMF by preventing the phosphorylation of I kappa B-alpha.
Tao
Qing-Song
QS
Department of General Surgery, Jinling Hospital, Clinical School of Nanjing University, Nanjing 210002, China. jan@medmail.com.cn.
Ren
Jian-An
JA
Li
Jie-Shou
JS
D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000008:administration & dosage / Q000302:isolation & purification / Q000494:pharmacology; D015153:Blotting, Western; D002478:Cells, Cultured; D018932:Chemokine CCL2 / Q000096:biosynthesis / Q000235:genetics; D018925:Chemokines / Q000096:biosynthesis / Q000235:genetics*; D003106:Colon / Q000166:cytology; D004224:Diterpenes / Q000008:administration & dosage / Q000302:isolation & purification / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D004852:Epoxy Compounds / Q000008:administration & dosage / Q000302:isolation & purification / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D020672:I-kappa B Proteins / Q000378:metabolism; D053583:Interleukin-1beta / Q000494:pharmacology*; D016209:Interleukin-8 / Q000096:biosynthesis / Q000235:genetics; D019278:Matrix Metalloproteinase 3 / Q000096:biosynthesis / Q000235:genetics*; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000378:metabolism; D010616:Phenanthrenes / Q000008:administration & dosage / Q000302:isolation & purification / Q000494:pharmacology*; D010766:Phosphorylation / Q000187:drug effects; D010946:Plants, Medicinal / Q000737:chemistry; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D029757:Tripterygium / Q000737:chemistry
D000894:Anti-Inflammatory Agents, Non-Steroidal; D018932:Chemokine CCL2; D018925:Chemokines; D004224:Diterpenes; D004852:Epoxy Compounds; D020672:I-kappa B Proteins; D053583:Interleukin-1beta; D016209:Interleukin-8; D016328:NF-kappa B; C000606204:NFKBIA protein, human; D010616:Phenanthrenes; D012333:RNA, Messenger; D000072000:NF-KappaB Inhibitor alpha; C001899:triptolide; D019278:Matrix Metalloproteinase 3