Estrogen-regulated expression of SK3 channel in rat colonic smooth muscle contraction

Life Sci. 2020 Dec 15:263:118549. doi: 10.1016/j.lfs.2020.118549. Epub 2020 Oct 9.

Abstract

Aims: Estrogen can induce inhibition of colonic smooth muscle contraction in male and female mice, which may lead to constipation; however, the mechanisms of inhibition are poorly understood. Hence, this study investigated the effect of estrogen on rat colonic smooth muscle contraction and role of small-conductance Ca2+-activated K+ 3 (SK3) and transcription factors (Sp1 and Sp3) in the underlying mechanisms.

Main methods: The experiment included 24 female Sprague-Dawley (SD) rats divided into 4 groups. The rats were oophorectomized surgically, and a silicone tube containing blank solvent, 0.3 mg/mL estrogen (E2), equal-concentration of estrogen and estrogen receptor antagonist (EI), and bovine serum albumin-E2 (BSA-E2) was implanted. The rats were sacrificed on day 14. The molecular insights were confirmed using real-time quantitative reverse transcription PCR (qRT-PCR) and western blot analyses to determine the effect of estrogenic stimulation on gene and protein expression analyses, respectively.

Key findings: The E2 group showed significantly greater SK3 expression (P < .005) compared with other groups and significantly lowers smooth muscle cell (SMC) contractility (P < .005). Estrogen stimulation and SK3 overexpression resulted in a significant decrease (P < .05) in Ca2+ mobilization in the E2 group versus the control group. Further, the E2 group showed significantly higher Sp1 mRNA (P < .05) but lower Sp3 mRNA expression (P < .05) and protein expression (P < .001) compared with other groups.

Significance: E2 may promote SK3 expression by its genomic effect and inhibit colonic contraction by affecting SK3 expression via an interaction between Sp1 and Sp3.

Keywords: Ca(2+); Colon; Estrogen; Myocyte; Small-conductance Ca(2+)-activated K(+) channels; Smooth muscle.

MeSH terms

  • Animals
  • Calcium / metabolism*
  • Cells, Cultured
  • Colon / cytology
  • Colon / drug effects
  • Colon / metabolism*
  • Estrogens / pharmacology*
  • Female
  • Gene Expression Regulation / drug effects*
  • Muscle Contraction*
  • Muscle, Smooth / cytology
  • Muscle, Smooth / drug effects
  • Muscle, Smooth / metabolism*
  • Rats
  • Rats, Sprague-Dawley
  • Small-Conductance Calcium-Activated Potassium Channels / genetics
  • Small-Conductance Calcium-Activated Potassium Channels / metabolism*
  • Sp1 Transcription Factor / genetics
  • Sp1 Transcription Factor / metabolism
  • Sp3 Transcription Factor / genetics
  • Sp3 Transcription Factor / metabolism

Substances

  • Estrogens
  • Kcnn3 protein, rat
  • Small-Conductance Calcium-Activated Potassium Channels
  • Sp1 Transcription Factor
  • Sp3 protein, rat
  • Sp3 Transcription Factor
  • Calcium