Inhibition of coagulation activation and inflammation by a novel Factor Xa inhibitor synthesized from the earthworm Eisenia andrei.
We have cloned an earthworm-derived Factor Xa (FXa) inhibitor, with an excellent inhibitory specificity from the midgut of the Eisenia andrei. We designate this inhibitor eisenstasin. An eisenstasin-derived small peptide (ESP) was synthesized and we examined whether ESP played an essential role in FXa inhibition. Compared to antistasin-derived small peptides (ASP) originating from leech, ESP primarily exhibited a high level of FXa inhibition in chromogenic peptide substrate assays and revealed an approximately 2-fold greater inhibition of FXa cleavage of a target protein than ASP. This suggests that ESP could be an effective anti-coagulant that targets FXa during the propagation step of coagulation. ESP also inhibited proteinase-activated receptor 2-mediated FXa activation, which may trigger endothelial inflammation. Endothelial nitric oxide (NO) was significantly reduced by ESP (p<0.0001), indicating that protease-activated receptor-2 (PAR-2) was effectively inactivated. We also found that ESP reduced the expressions of pro-inflammatory cytokines (IL-1alpha, IL-1beta, IL-8, IL-16, MCP-1, MIP-1alpha and MIP-1beta) by cultured cells treated with both ESP and FXa. Our results provide the first evidence that ESP might interrupt coagulation cascades by inhibiting FXa, and thereby may effectively control the bidirectional alternation between coagulation and inflammation.
Joo
Seong Soo
SS
Research Institute of Veterinary Medicine, Chungbuk National Universit, Cheongju, Republic of Korea.
Won
Tae Joon
TJ
Kim
Jong Sung
JS
Yoo
Yeong Min
YM
Tak
Eun Sik
ES
Park
So-Young
SY
Park
Hee Yong
HY
Hwang
Kwang Woo
KW
Park
Soon Cheol
SC
Lee
Do Ik
DI
D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000138:chemical synthesis / Q000494:pharmacology*; D000925:Anticoagulants / Q000138:chemical synthesis / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000494:pharmacology; D015153:Blotting, Western; D002460:Cell Line; D016207:Cytokines / Q000096:biosynthesis; D004354:Drug Screening Assays, Antitumor; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D015951:Factor Xa / Q000494:pharmacology; D065427:Factor Xa Inhibitors; D006801:Humans; D007447:Invertebrate Hormones / Q000138:chemical synthesis / Q000494:pharmacology*; D009569:Nitric Oxide / Q000378:metabolism; D009835:Oligochaeta / Q000378:metabolism*; D010766:Phosphorylation; D044463:Receptor, PAR-1 / Q000096:biosynthesis / Q000235:genetics
D000894:Anti-Inflammatory Agents, Non-Steroidal; D000925:Anticoagulants; D000970:Antineoplastic Agents; D016207:Cytokines; D065427:Factor Xa Inhibitors; D007447:Invertebrate Hormones; D044463:Receptor, PAR-1; C539019:eisenstasin; D009569:Nitric Oxide; D048049:Extracellular Signal-Regulated MAP Kinases; D015951:Factor Xa