Effect of Glyphaea brevis twigs extract on cell viability, apoptosis induction and mitochondrial membrane potential in TM3 Leydig cells

Andrologia. 2019 Aug;51(7):e13312. doi: 10.1111/and.13312. Epub 2019 May 15.

Abstract

Glyphaea brevis twigs (Spreng) Monach. (GBT) are used by local herb healers to manage male sexual fertility disorders. The aim of this study was to examine the effects of G. brevis twigs on TM3 Leydig cells. GBT was extracted using methanol solvent, and Leydig cells were exposed to the respective concentrations of GBT extract (0.1, 1, 10, 100 and 1,000 μg/ml) for 24 and 72 hr respectively. Parameters evaluated include cell morphology, viability (MTT assay), mitochondrial membrane potential (TMRE dye), apoptosis (Annexin V Alexa Fluor 488 binding) and RT-qPCR analyses of the mRNA expression. Results revealed that GBT had no cytotoxic effect on cell viability and the cell morphology. GBT also revealed a considerable elevation (p < 0.05) in fluorescence intensity, accompanied by intact mitochondria in TM3 Leydig cells. Furthermore, GBT resulted in the reduction of necrotic and apoptotic cells. The mRNA StAR was upregulated markedly with the effect prominent at 100 μg/ml. This study showed that GBT might be useful for managing male infertility ailments.

Keywords: Glyphaea brevis twigs; TM3 Leydig cells; apoptosis; mRNA.

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Cell Line
  • Cell Survival / drug effects
  • Infertility, Male / drug therapy
  • Leydig Cells / drug effects*
  • Leydig Cells / metabolism
  • Male
  • Malvaceae / chemistry*
  • Membrane Potential, Mitochondrial / drug effects
  • Methanol / chemistry
  • Methanol / isolation & purification
  • Mice
  • Mitochondria / drug effects
  • Phosphoproteins / metabolism*
  • Plant Components, Aerial / chemistry
  • Plant Extracts / isolation & purification
  • Plant Extracts / pharmacology*
  • Plant Extracts / therapeutic use
  • Up-Regulation

Substances

  • Phosphoproteins
  • Plant Extracts
  • steroidogenic acute regulatory protein
  • Methanol