Discovery of a novel megakaryopoiesis enhancer, ingenol, promoting thrombopoiesis through PI3K-Akt signaling independent of thrombopoietin.

Thrombocytopenia, a most common complication of radiotherapy and chemotherapy, is an important cause of morbidity and mortality in cancer patients. However, there are still no approved agents for the treatment of radiation- and chemotherapy-induced thrombocytopenia (RIT and CIT, respectively). In this study, a drug screening model for predicting compounds with activity in promoting megakaryocyte (MK) differentiation and platelet production was established based on machine learning (ML), and a natural product ingenol was predicted as a potential active compound. Then, in vitro experiments showed that ingenol significantly promoted MK differentiation in K562 and HEL cells. Furthermore, a RIT mice model and c-MPL knock-out (c-MPL-/-) mice constructed by CRISPR/Cas9 technology were used to assess the therapeutic action of ingenol on thrombocytopenia. The results showed that ingenol accelerated megakaryopoiesis and thrombopoiesis both in RIT mice and c-MPL-/- mice. Next, RNA-sequencing (RNA-seq) was carried out to analyze the gene expression profile induced by ingenol during MK differentiation. Finally, through experimental verifications, we demonstrated that the activation of PI3K/Akt signaling pathway was involved in ingenol-induced MK differentiation. Blocking PI3K/Akt signaling pathway abolished the promotion of ingenol on MK differentiation. Nevertheless, inhibition of TPO/c-MPL signaling pathway could not suppress ingenol-induced MK differentiation. In conclusion, our study builds a drug screening model to discover active compounds against thrombocytopenia, reveals the critical roles of ingenol in promoting MK differentiation and platelet production, and provides a promising avenue for the treatment of RIT.

The phytochemistry, pharmacokinetics, pharmacology and toxicity of Euphorbia semen.

ETHNOPHARMACOLOGICAL RELEVANCE: Euphorbia semen, the dried and ripe seed of Euphorbia lathyris Linnaeus, is widely cultivated for traditional medicine use. This semen is used to expel water, help with phlegm retention, promote blood circulation, remove blood stasis, cure tinea and scabies, and treat amenorrhea, snakebites, terminal schistosomiasis, anuria and constipation. AIM OF THE REVIEW: This review provides updated, comprehensive and categorized information on the local and traditional uses, phytochemistry, pharmacokinetics, pharmacological activities and toxicity of Euphorbia semen. Future research to deepen the recognition and utilization of Euphorbia semen is proposed. MATERIALS AND METHODS: This article conducted a literature review on information about Euphorbia semen in multiple Internet databases, including PubMed, Web of Science, Wiley, Science Direct, Elsevier, ACS publications, SciFinder, Google Scholar and China National Knowledge Internet, until March of 2018. In this manuscript, a number of books, PhD and MSc dissertations, and Chinese Pharmacopeia were also used as references. RESULTS: Approximately 240 chemical constituents have been isolated and identified from Euphorbia semen, namely, diterpenoids, coumarins, flavonoids, fatty acids, amino acids, and steroids. Pharmacokinetic study focused on investigating absorption, distribution, metabolism and excretion (ADME). The chemical constituents have extensive pharmacological effects, such as diuresis and anti-hyperuricaemia, anti-inflammation, antiviral, anticancer, antioxidant, antipigmentation, anti-platelet aggregation and anti-allergic activities, as well as hepatoprotection and neuroprotection. The toxicity of Euphorbia semen, including acute toxicity, target organ irritation and cocareinogenic effects, have been reported, and the detoxification methods are reviewed. CONCLUSION: Euphorbia semen has extensive pharmacological activity and excellent clinical value, along with intense intestinal irritation. Although plenty of chemical constituents have been isolated and identified, the exact pharmacological and toxicological mechanisms still need to be explored.