title: Salvianolic acid B maintained stem cell pluripotency and increased proliferation rate by activating Jak2-Stat3 combined with EGFR-Erk1/2 pathways.
abstract: Embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) are considered the most powerful in terms of differentiating into three-germ-layer cells. However, maintaining self-renewing ESCs and iPSCs in vitro requires leukemia-induced factor (LIF), an expensive reagent. Here we describe a less expensive compound that may serve as a LIF substitute-salvianolic acid B (Sal B), a Salvia miltiorrhiza extract. We found that Sal B is capable of upregulating Oct4 and Sox2, two genes considered important for the maintenance of ESC pluripotency. Our MTT data indicate that instead of triggering cell death, Sal B induced cell proliferation, especially at optimum concentrations of 0.01 nM and 0.1 nM. Other results indicate that compared to non-LIF controls, Sal B-treated ESCs expressed higher levels of several stem cell markers while still maintaining differentiation into three-germ-layer cells after six passages. Further, we found that Sal B triggers the Jak2-Stat3 and EGFR-ERK1/2 signaling pathways. Following Sal B treatment, (a) levels of phosphorylated (p)-Jak2, p-Stat3, p-EGFR, and p-ERK proteins all increased; (b) these increases were suppressed by AG490 (a Jak2 inhibitor) and ZD1839 (an EGFR inhibitor); and (c) cytokines associated with the Jak2-Stat3 signaling pathway were upregulated. Our findings suggest that Sal B can be used as a LIF replacement for maintaining ESC pluripotency while increasing cell proliferation.
authors.affiliation: Graduate Institute of Immunology, China Medical University, Taichung, Taiwan.
authors.forename: Chia Hui
authors.initials: CH
authors.lastname: Liu
authors.forename: Woei-Cherng
authors.initials: WC
authors.lastname: Shyu
authors.forename: Ru-Huei
authors.initials: RH
authors.lastname: Fu
authors.forename: Shyh-Jer
authors.initials: SJ
authors.lastname: Huang
authors.forename: Cheng-Hsuan
authors.initials: CH
authors.lastname: Chang
authors.forename: Yu-Chuen
authors.initials: YC
authors.lastname: Huang
authors.forename: Shih-Yin
authors.initials: SY
authors.lastname: Chen
authors.forename: Shinn-Zong
authors.initials: SZ
authors.lastname: Lin
authors.forename: Shih-Ping
authors.initials: SP
authors.lastname: Liu
mesh_terms: D000818:Animals; D001572:Benzofurans / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D066246:ErbB Receptors / Q000378:metabolism*; D053614:Janus Kinase 2 / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D055748:SOXB1 Transcription Factors / Q000378:metabolism; D050796:STAT3 Transcription Factor / Q000378:metabolism; D027561:Salvia miltiorrhiza / Q000737:chemistry / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D053630:Depsides
chemical_list: D001572:Benzofurans; D066246:ErbB Receptors; D053614:Janus Kinase 2; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D050814:Octamer Transcription Factor-3; D055748:SOXB1 Transcription Factors; D050796:STAT3 Transcription Factor; C076944:salvianolic acid B; D053630:Depsides; C541485:EGFR protein, mouse