Mitochondria-dependent apoptogenic activity of the aqueous root extract of Croton membranaceus against human BPH-1 cells.
Croton membranaceus aqueous root extract (CMARE) is among the widely used phytotherapeutics in Ghana for the management of benign prostatic hyperplasia (BPH) and prostate cancer. However, the mechanism of action of CMARE remains to be elucidated. This study aimed to establish whether apoptosis is involved in the antiproliferative effect of CMARE on human BPH-1 cells. We determined the effect of treatment with 0, 1, 3, and 5 mg/mL CMARE for 24, 48, and 72 h on the viability and morphology of BPH-1 cells using the MMT assay and phase-contrast microscopy, respectively. We examined the apoptosis-inducing effects of CMARE after 48 h at the cellular level using Hoescht 33258 and JC-1 dye staining and flow cytometry analysis. We performed reverse transcription polymerase chain reaction and Western blotting to confirm the apoptotic effects of CMARE at the molecular level. CMARE induced a significant dose-dependent inhibition in the proliferation of BPH-1 cells (P < 0.05) and an alteration in their morphology and a reduction their density. Furthermore, CMARE induced dose-dependent staining of the nuclear chromatin, significant DNA fragmentation with G₀/G₁ sub-diploid cells (P < 0.01), and loss of the mitochondrial membrane potential in the treated cells compared to the controls after 48 h (P < 0.01). Additionally, while CMARE induced a significant upregulation of the mRNA and protein levels of Bax, those of Bcl2 did not change significantly. Therefore, induction of mitochondria-dependent apoptosis of BPH-1 cells may be a possible mechanism of action of CMARE.
Academy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, China.
D K
DK
Afriyie
Department of Medical Laboratory Sciences, School of Allied Health Sciences, Accra, Ghana gasare@chs.edu.gh.
G A
GA
Asare
Department of Pharmacology, University of Ghana Medical School, Accra, Ghana.
K
K
Bugyei
Academy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, China.
J
J
Lin
Academy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, China.
J
J
Peng
Academy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, China.
Z
Z
Hong
D017209:Apoptosis / Q000187:drug effects*; D006690:Bisbenzimidazole; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D053144:Cell Nucleus Shape / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D029802:Croton / Q000737:chemistry*; D005434:Flow Cytometry; D006801:Humans; D008297:Male; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D010936:Plant Extracts / Q000494:pharmacology*; D018517:Plant Roots / Q000737:chemistry*; D011470:Prostatic Hyperplasia / Q000473:pathology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism
D010936:Plant Extracts; D019253:Proto-Oncogene Proteins c-bcl-2; D006690:Bisbenzimidazole