Lipopolysaccharide Induces Interleukin-8 Production By Human Cervical Smooth Muscle Cells

Abstract

Objective: We examined the effect of lipopolysaccharide (LPS), a component of the outer wall of gram-negative bacteria, on expression of the neutrophil chemoattractant interleukin-8 (IL-8) and the effects of IL-8 treatment on release of matrix metabolizing enzymes in human cervical smooth muscle cells (CSMCs).

Methods: Human CSMCs were exposed to Escherichia coli LPS, and the expression of IL-8 mRNA was analyzed by Northern blotting. The Il-8 promoter activity was examined by dual luciferase assay, and the IL-8 concentration was assessed by enzyme-linked immunosorbent assay. We also treated the CSMCs with human IL-8 and examined the expression of matrix-degrading enzymes.

Results: E coli LPS (100 ng/mL) increased the expression of IL-8 mRNA 12.8-fold after 3 hours. This up-regulation was maintained for up to 24 hours. Lipopolysaccharide treatment produced a fivefold increase in IL-8 promoter activity in CSMCs transfected with an Il-8 promoter-reporter construct. IL-8 concentrations in conditioned medium of CSMC cultures treated with E coli LPS increased approximately 18-fold compared with the control culture. Northern blot analysis and zymography revealed that exogenous human IL-8 had no significant effect on the expression of matrix metalloproteinase (MMP)-1, -3, and tissue inhibitor of metalloproteinase (TIMP)-1 mRNAs, and on the secretion MMP-2 and -9 in CSMCs.

Conclusion: We conclude that CSMCs respond to LPS with increased expression of IL-8 mRNA and secreted IL-8, and that expression of matrix metabolizing enzymes in CSMCs is not directly affected by IL-8. IL-8 produced by CSMCs in reponse to gram-negative infection may promote neutrophil invasion, and release of neutrophil matrix-degrading enzymes may participate in the matrix remodeling associated with parturition.

Keywords

Bacterial vaginosis cervical ripening human cervical smooth muscle cells interleukin-8 lipopolysaccharide

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References

1. Liggins GC . Cervical ripening as an inflammatory reaction. In: Ellwood DA , Anderson ABM , eds. The cervix in pregnancy and labour. Edinburgh, UK: Churchill Livingstone, 1981:1-9.

2. Thomson AJ , Telfer JF , Young A , Campbell S , Stewart CJR , Cameron IT , Greer IA , Norman JE . Leukocytes infiltrate the myometrium during human parturition: Further evidence that labor is an inflammatory process. Hum Reprod 1999;14:229-236.

3. Winkler M , Fischer DC , Hlubek M , van de Leur E , Haubeck HD , Rath W . Interleukin-1beta and interleukin-8 concentrations in the lower uterine segment during parturition at term. Obstet Gynecol 1998;91:945-949.

4. Sennström M , Ekman G , Westergren-Thorsson G , et al. Human cervical ripening, an inflammatory process mediated by cytokines. Mol Hum Reprod 2000;6:375-381.

5. Watari M , Watari H , DiSanto ME , Chacko S , Shi GP , Strauss JF III . Pro-inflammatory cytokines induce expression of matrixmetabolizing enzyme in human cervical smooth muscle cells. Am J Pathol 1999;154:1755-1762.

6. Junqueira LC , Zugaib M , Montes GS , Toledo OMS , Krisztan RM , Shigihara KM . Morphologic and histochemical evidence for the opccurrence of the collagenolysis and for the role of neutrophilie polymorphonuclear leukocytes during cervical dilatation. Am J Obstet Gynecol 1980;138:273-281.

7. Rajabi M , Dean DD , Beydoun SN , Woessner JF Jr . Elevated tissue levels of collagenase during dilatation of the uterine cervixin human parturition. Am J Obstet Gynecol 1980;138:273-281.

8. Uldbjerg N , Ekman G , Malmstrom A , Olsson K , Ulmsten U . Ripening of the human uterine cervix related changes in collagen, glycosaminoglycans, and collagenolytic activity. Am J Obstet Gynecol 1983;147:662-666.

9. Leppert PC , Yu SY , Keller S , Cerreta J , Mand I . Decreased elastic fibers and desmosine content in competent cervix. Am J Obstet Gynecol 1987;157:1134-1139.

10.

10. Rotten D , Gavignet C , Colin MC , Robert AM , Godeau G . Evollution of the elastic fiber network of the human uterine cervix before, during and after pregnancy. A quantitative evaluation by automated image analysis. Clin Physiol Biochem 1988;6:285-292.

11.

11. Baggliolini M , Walz A , Kunkel SL , Neutrophil attractant/activating peptide-1/interleukin-8: A novel cytokine that attracts neutrophils. J Clin Invest 1989;84:1045-1049.

12.

12. Williems J , Joniau M , Cinque S , Damme JV . Human granulocyte chemotactic peptide (IL-8) as a specific neutrophil degranulator: Comparison with other cytokines. Immunology 1989;67:540-542.

13.

13. Osmer RGW , Blaser J , Kuhn W , Tschesche H . Interleukin-8 synthesis and the onset of labor. Obstet Gynecol 1995;86:223-229.

14.

14. Romero R , Ceska M , Avila C , Mazor M , Behnke E , Lindley I . Neutrophil attractant/activating peptide-1/interleukin-8 in term and preterm patrurition. Am J Obstet Gynecol 1999;165:813-820.

15.

15. Winkler M , Fischer DC , Ruck P , et al. Parturition at term: Parallel increases in interleukin-8 and proteinase concentrations and neutrophil count in the lower uterine segment. Hum Reprod 1999;14:1096-1100.

16.

16. Hitti J , Hiller LS , Agnew KJ , Krohn MA , Reisner DP , Eschenbach DA . Vaginal indicators of amniotic fluid infection in preterm labor. Obstet Gynecol 2001;97:211-219.

17.

17. Barcley CG , Brennand JE , Kelly RW , Calder AA . Interleukin-8 production by the human cervix. Am J Obstet Gynecol 1993;169:625-632.

18.

18. Kelly RW , Leask R , Calder AA . Choriodecidua production of interleukin-8 and mechanism of parturition. Lancet 1992;339:776-777.

19.

19. Chwalisz K , Benson M , Scholz P , Daum JM , Beier H , Hegele-Hartung C . Cervical ripening with the cytokines interleukin-8, interleukin-1 beta and tumor necrosis factor alpha in guinea-pigs. Hum Reprod 1994;9:2173-2181.

20.

20. ElMaradny E , Kanayama N , Halim A , Machara K , Sumitomo K , Terao T . Interleukin-8 induces cervical ripening in rabbits. Am J Obstet Gynecol 1994;171:77-83.

21.

21. Luca M , Huang S , Gershenwald JE , Singh RK , Reich R , Bar-Eli M . Expression of interleukin-8 by human melanoma cells upregulates MMP-2 activity and increases tumor growth and metastasis. Am J Pathol 1997;151:1105-1113.

22.

22. Xu L , Fidler IJ . Interlukin 8: An autocrine growth factor for human ovarian cancer. Oncol Res 2000;12:97-106.

23.

23. Massod R , Cai J , Tulpule A , et al. Interleukin-8 is an autocrine growth factor and surrogate marker for Kaposi's sarcoma. Clin Cancer Res 2001;8:2693-2702.

24.

24. Wang Z , Castresana MR , Newman WH . NF-kappaB is required for TNF-alpha-directed smooth muscle cell migration. FEBS Lett 2001;23:360-364.

25.

25. Hatthachote P , Gillespie JI . Complex interactions between sex steroids and cytokines in the human pregnant myometrium: Evidence for the autocrine signaling system at term. Endocrinology 1999;140:2533-2540.

26.

26. Raetz CRH , Ulevitch RJ , Wright SD , Sibley CH , Ding A , Nathan CF . Gram-negative endotoxin: An extraordinary lipid with profound effects on eukaryotic signal transduction. FASEB J 1991;5:2652-2660.

27.

27. Raabe T , Bukrinsky M , Alexander Currie R . Relative contribution of transcription and translation to the induction of tumor necrosis factor-alpha by lipopolysaccharide. J Biol Chem 1998;273:974-980.

28.

28. Seiler-tuyns A , Dufour N , Spertini F . Human tumor necrosis factor-A gene 3′ untranslated region confers inducible toxin responsiveness to homologous promoter in monocytic THP-1 cells. J Biol Chem 1999;274:21714-21718.

29.

29. Brown Z , Gerritsen ME , Carley WW , Strieter RM , Kunkel SL , Westwick J . Chemokine gene expression and secretion by cytokine-activated human microvascular endothelial cells. Differential regulation of monocyte chemoattractant protein-1 and interleukin-8 in response to interferon-γ. Am J Pathol 1994;145:913-921.

30.

30. Kaga N , Katsuki Y , Obata M , Shibutani Y . Repeated administration of low-dose lipopolysaccharide induces preterm delivery in mice: A model for human preterm parturition and for assessment of the therapeutic ability of drugs against preterm delivery. Am J Obstet Gynecol 1996;174:754-759.

31.

31. Watari M , Watari H , Nachampkin I , Strauss JF III . Lipopolysac-charide induces the expression of genes encoding pro-inflammatory cytokines and elastin-degrading enzyme, cathepsin S, in human cervical smooth muscle cells. J Soc Gynecol Investig 2000;7:190-198.

32.

32. Winkler M , Rath W , Fischer DC , van de Leur E , Haubeck HD . Regulation of interleukin-8 synthesis in human lower uterine segment fibroblasts by cytokines and growth factors. Obstet Gynecol 2000;95:584-588.

33.

33. Onodera S , Kaneda K , Mizue Y , Koyama Y , Fujinaga M , Nishihira J . Macrophage migration inhibitory factor up-regulates expression of matrix metalloproteinases in synovial fibroblasts of rheumatoid arthritis. J Biol Chem 2000;275-444-450.

34.

34. Pugin J , Widmer MC , Kossodo S , Liang CM , Preas HL II , Suffredini AF . Human neutrophils secrete gelatinase B in vitro and in vivo in response to endotoxin and proinflammatory mediators. Am J Respir Cell Mol Biol 1999;20:458-464.

35.

35. Helmer H , Tretzmuller U , Brunbauer M , Kaider A , Husslein P , Knöfler M . Production of oxytocin receptor and cytokines in primary uterine smooth muscle cells cultivated under inflammatory conditions. J Soc Gynecol Investig 2002;9:15-21.

36.

36. Moser B , Barella L , Mattei S , et al. Expression of transcripts for two interleukin-8 receptors in human phagocytes, lymphocytes and melanoma cells. Biochem J 1993;294:285-292.

37.

37. Belperio JA , Keane MP , Arenberg DA , et al. CXC chemokines in angiogenesis. J Leukocyte Biol 2000;68:1-8.

38.

38. Angus SR , Segel SY , Hsu CD , et al. Amniotic fluid matrix metalloproteinase-8 indicates intra-amniotic infection. Am J Obster Gynecol 2001;185-1232-1238.

39.

39. Yue TL , Wang X , Sung CP , Olson B , McKenna PJ , Gu JL , Feuerstein GZ . Interleukin-8. A mitogen and chemoattractant for vascular smooth muscle cells. Circ Res 1994;75:1-7.

40.

40. Iwabe T , Harada T , Tsudo T , Tanikawa M , Onohara Y , Terakawa N . Pathogenetic significance of increased levels of interleukin-8 in the peritoneal fluid of patients with endometriosis. Fertil Steril 1998;69:924-930.

41.

41. Lin G , Pearson AE , Scamurra Rw , et al. Regulation of interleukin-8 expression in porcine alveolar macrophages by bacterial lipopolysaccharide. J Biol Chem 1994;206:77-85.

42.

42. Elliott CL , Allport VC , Loudon JAZ , Wu Gd , Bennett PR . Nuclear factor-kappa B is essential for up-regulation of interleukin-8 expression in human amnion and cervical epithelial cells. Mol Hum Reprod 2001;7:787-790.

43.

43. Gesser B , Deleuran B , Lund M et al. Interleukin-8 induces its own production in CD4+ T lymphocytes: A process regulated by IL-10. Biochem Biophys Res Commun 1995;210:660-669.