Virodhamine, an endocannabinoid, induces megakaryocyte differentiation by regulating MAPK activity and function of mitochondria

Abstract

Endocannabinoids are well-known regulators of neurotransmission by activating the cannabinoid (CB) receptors. Endocannabinoids are being used extensively for the treatment of various neurological disorders such as Alzheimer's and Parkinson's diseases. Although endocannabinoids are well studied in cell survival, proliferation, and differentiation in various neurological disorders and several cancers, the functional role in the regulation of blood cell development is less examined. In the present study, virodhamine, which is an agonist of CB receptor-2, was used to examine its effect on megakaryocytic development from a megakaryoblastic cell. We observed that virodhamine increases cell adherence, cell size, and cytoplasmic protrusions. Interestingly, we have also observed large nucleus and increased expression of megakaryocytic marker (CD61), which are the typical hallmarks of megakaryocytic differentiation. Furthermore, the increased expression of CB2 receptor was noticed in virodhamine-induced megakaryocytic cells. The effect of virodhamine on megakaryocytic differentiation could be mediated through CB2 receptor. Therefore, we have studied virodhamine induced molecular regulation of megakaryocytic differentiation; mitogen-activated protein kinase (MAPK) activity, mitochondrial function, and reactive oxygen species (ROS) production were majorly affected. The altered mitochondrial functions and ROS production is the crucial event associated with megakaryocytic differentiation and maturation. In the present study, we report that virodhamine induces megakaryocytic differentiation by triggering MAPK signaling and ROS production either through MAPK effects on ROS-generating enzymes or by the target vanilloid receptor 1-mediated regulation of mitochondrial function.

Low molecular weight heparin treatment reduced apoptosis and oxidative cytotoxicity in the thrombocytes of patients with recurrent pregnancy loss and thrombophilia: Involvements of TRPM2 and TRPV1 channels

“…In addition to the stimulation of [Ca 2+ ] i increase dependent-caspase-3 and caspase-9 pathways of apoptosis, [Ca 2+ ] i signal via the stimulation of TRPM2 and TRPV1 was found to alter the MiMP that might be critical for the induction of apoptosis. 12,[24][25][26]34,35 Consistently, the results in the current study revealed that RPL increase MiMP levels, which was found to be attributable to the changes in the [Ca 2+ ] i concentration as evidenced by the preventive actions of ACA and CPZ, antagonists of TRPM2 and TRPV1. Our recent results including others have indicated that increased cytROS as well as [Ca 2+ ] i are the potent inducers of MiMP level that triggers the mitochondrial pathway of apoptosis by upregulating the thrombocyte active caspase-3 and caspase-9.…”

“…The inhibition of RPL‐induced increase of caspase‐3, and caspase ‐9 but the decrease of cell viability by the treatments of ACA and CPZ as well as the LMWH treatment suggested the important role of [Ca 2+ ] i , TRPM2, and TRPV1 in the induction of apoptosis. In addition to the stimulation of [Ca 2+ ] i increase dependent‐caspase‐3 and caspase‐9 pathways of apoptosis, [Ca 2+ ] i signal via the stimulation of TRPM2 and TRPV1 was found to alter the MiMP that might be critical for the induction of apoptosis 12,24–26,34,35 . Consistently, the results in the current study revealed that RPL increase MiMP levels, which was found to be attributable to the changes in the [Ca 2+ ] i concentration as evidenced by the preventive actions of ACA and CPZ, antagonists of TRPM2 and TRPV1.…”

“…High levels of TRPM2 and TRPV1 activation were found in the megakaryocytes 21,22 . Increased oxidative toxicity causes apoptosis in megakaryocytes and leucocytes by activating calcium channels such TRPM2, TRPV1, and voltage‐gated calcium channels (VGCC) through a feedback mechanism; elevated cytROS can worsen the stimulation of apoptosis via the activation of the channels 23–25 . However, the negative apoptotic and oxidative effects in the cells of abortion patients were reduced by the therapy with antioxidants, which inhibited the TRPM2 and TRPV1 26–28 .…”

“…21,22 Increased oxidative toxicity causes apoptosis in megakaryocytes and leucocytes by activating calcium channels such TRPM2, TRPV1, and voltagegated calcium channels (VGCC) through a feedback mechanism; elevated cytROS can worsen the stimulation of apoptosis via the activation of the channels. [23][24][25] However, the negative apoptotic and oxidative effects in the cells of abortion patients were reduced by the therapy with antioxidants, which inhibited the TRPM2 and TRPV1. [26][27][28] Patients with reproductive abnormalities are treated with LMWH, and this medication has an antioxidant effect by increasing the levels of GSH and GSH-Px in the patients' blood.…”

Emerging Significance and Therapeutic Potential of Extracellular vesicles

“…Of course, EVs also contain nucleic acids, including mRNA, miRNA, DNA, and mitochondrial DNA (mtDNA). These nucleic acids are mainly involved in inflammation and as diagnostic biomarkers in tumors 25 , 26 . In recent years, scientists have paid more attention to EVs due to their key roles in biological systems.…”