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  • Title: Relaxin stimulates interstitial collagenase activity in cultured uterine cervical cells from nonpregnant and pregnant but not immature guinea pigs; estradiol-17 beta restores relaxin's effect in immature cervical cells.
    Author: Mushayandebvu TI, Rajabi MR.
    Journal: Biol Reprod; 1995 Nov; 53(5):1030-7. PubMed ID: 8527505.
    Abstract:
    Relaxin has been implicated in dilatation of the cervix at parturition. Dilatation of the guinea pig cervix at parturition is mediated by an estrogen-induced degradation of type I collagen by interstitial collagenase (matrix metalloproteinase I, MMPI). This study was designed to test the hypothesis that human recombinant relaxin (hrR) stimulates MMPI activity in cultured guinea pig cervical cells. Primary cervical monolayer cultures from immature, adult nonpregnant, and 50-day-pregnant Hartley guinea pigs were exposed to hrR (1-1000 ng/ml) daily for 3 days in serum-free Dulbecco's Modified Eagle's Medium (DMEM). The effect of priming immature cells with estradiol-17 beta (E2, 10(-6) M) daily for 3 days prior to treatment with hrR was also determined. Tissue inhibitors of metalloproteinases were inactivated by reduction, alkylation, and dialysis. MMPI activity at 96 h was assayed via a highly sensitive and specific assay that utilizes [3H]telopeptide-free type I collagen as a substrate. Aminophenylmercuric acetate (0.5 mM) was used to activate latent MMPI. Phenanthroline-1,10 (1 mM), a known inhibitor of metalloproteinases, was used as a blank control. Phorbol-12-myristate-13-acetate (PMA, 10(-8) M), a known stimulator of MMPI biosynthesis, was used as a positive control. The hrR in serum-free DMEM had no significant effect on cell number in nonpregnant, 50-day-pregnant, E2-primed, or nonprimed immature animals. It stimulated MMPI activity in a dose-dependent manner with a maximum response at 10 ng/ml in nonpregnant (2-fold), 50-day-pregnant (3-fold), and E2-primed immature (3-fold) animals.(ABSTRACT TRUNCATED AT 250 WORDS)
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