Developmental changes in the inhibition of cultured rat uterine cell proliferation by opioid peptides

Abstract

Opioid peptides are negative regulators of cell proliferation in several organs including the uterus. In the present study, the ontogeny of the direct inhibitory action of opioid peptides on the proliferation of cultured rat uterine cells was investigated. Uteri of 7, 14, 21, 28, 35 and 60-day-old rats were removed in a sterile way. Tissue blocks were dispersed by limited digestions with trypsin and collagenase. Cells were cultured in enriched Dulbecco's modified Eagle's medium (DMEM). Treatments were present during the entire culture period. Cell densities of the monolayers were determined by counting the cells following trypsinization and trypan blue exclusion. Rat uterine mixed cell cultures grew to confluence within 10 days. The average population doubling time gradually increased with the age of animals. Epidermal growth factor (EGF) increased cell densities of cultures from all age groups. The oestradiol (E2)-responsiveness appeared at 21 days of age. The effect of [D-Met2-Pro5]-enkephalinamide (ENK) was biphasic. ENK and [Met5]-enkephalin (OGF) decreased cell densities of both unstimulated and EGF-stimulated cultures from 7-day-old rats to the same extent. ENK failed to act in 14-day-old animals. From 21 days of age on, the E2- or EGF-stimulated proliferation was inhibited only by ENK and DAMGO, while 30 nm DPDPE, Dynorhin-A, OGF, [Leu5]-enkephalin, beta-endorphin, and morphiceptin were ineffective. The half-inhibitory concentration of ENK was 0.3 nm. The effects of ENK were prevented by concomitant treatment with naloxone. Our novel data demonstrate two different phases of the inhibitory action of opioid peptides on rat uterine cell proliferation during ontogeny with an insensitive interval in between.

Delta opioid peptide DADLE and naltrexone cause cell cycle arrest and differentiation in a CNS neural progenitor cell line

“…The antiproliferative effect seen with DADLE and naltrexone is certainly nonopioid in nature. Opioids have previously been shown to exert proliferative or antiproliferative effect via an opioid or nonopioid action (Ilyinsky et al, 1987; Kornyei et al, 2003; Martin-Kleiner, 2002; Persson et al, 2003a,b; Zagon and McLaughlin, 2003; McLaughlin et al, 2003). Our results showing the unidirectional action of DADLE and naltrexone on AF5 cells strengthen the notion of the nonopioid nature of certain actions of opioid peptides in cellular systems.…”

The OGF–OGFr Axis Utilizes the p16INK4aand p21WAF1/CIP1Pathways to Restrict Normal Cell Proliferation

“…Because OGF depresses DNA synthesis and subsequent cell/tissue growth in a wide variety of normal and developing cells in humans and animals, including ectodermal, mesodermal, and endodermal derivatives Hauser and Stiene-Martin, 1991;Isayama et al, 1991;Zagon and McLaughlin, 1991;Zagon et al, 1994Zagon et al, , 1995bZagon et al, , 1996aZagon et al, ,b, 1997Zagon et al, , 1999bMcLaughlin, 1996;Vertes et al, 1996;McLaughlin and Wu, 1998;Blebea et al, 2000Blebea et al, , 2002Wilson et al, 2000;Kornyei et al, 2003), the question arises as to the mechanism of peptide action on the cell cycle in these cells. The present investigation examined the specific target(s) in the cell cycle for the OGF-OGFr axis in cells derived from four normal human tissues: umbilical vein endothelial This article was published online ahead of print in MBC in Press (http://www.molbiolcell.org/cgi/doi/10.1091/mbc.E08 -07-0681) on October 15, 2008. cells (HUVECs), epidermal keratinocytes (NHEKs), dermal fibroblasts (NHDFs), and mesenchymal stem cells (hMSCs).…”

“…OGF is a constitutively expressed native opioid that is autocrine produced and secreted to inhibit the growth of both normal cells McLaughlin, 1987, 1991;Hauser et al, 1990;Stiene-Martin and Hauser, 1990;Hauser and Stiene-Martin, 1991;Isayama et al, 1991;Villiger and Lotz, 1992;Zagon et al, 1994Zagon et al, , 1995aZagon et al, ,b, 1996aZagon et al, , 1997aZagon et al, ,b, 1998Zagon et al, , 1999bZagon et al, , 2000bMcLaughlin, 1996;Vertes et al, 1996;McLaughlin and Wu, 1998;Blebea et al, 2000;Wilson et al, 2000;Kornyei et al, 2003;Robertson and Andrew, 2003;Malendowicz et al, 2005) and cancer cells (Zagon et al, 1996c;Cheng et al, 2007Cheng et al, , 2008. The action of OGF is tonic, stereospecific, reversible, noncytotoxic, and nonapoptotic inducing, is not associated with differentiative, migratory, invasive, or adhesive processes, is independent of serum, anchorage-independent and occurs at physiologically relevant concentrations (Zagon et al, , 2007a, 2004.…”

Influence of the Preparation «dalargin» on the Condition of Rat’s Postpartum Uterus

“…Большое количество морфологических исследований, касающихся послеродовых изменений в матке, посвящено структурным преобразованиям эндометрия [4,5] и, в меньшей степени, инволютивным процессам в миометрии [2,7]. Имеющиеся в литературе отдельные морфологические и биохимические исследования процессов, происходящих в миометрии послеродовой матки, не формируют целостного представления о роли опиоидных пептидов в процессах его физиологической послеродовой инволюции [6,9]. В связи с этим, изучение влияния опиоидных пептидов на процессы дифференцировки и пролиферации клеток эндометрия и миометрия матки является актуальной проблемой современной медицинской науки.…”