Mesenchymal stem cells (MSCs) have shown great potential for the treatment of liver injuries, and the therapeutic efficacy greatly depends on their homing to the site of injury. In the present study, we detected significant upregulation of hepatocyte growth factor (HGF) in the serum and liver in mice with acute or chronic liver injury. In vitro study revealed that upregulation of miR-9-5p or miR-221-3p promoted the migration of human MSCs (hMSCs) toward HGF. Moreover, overexpression of miR-9-5p or miR-221-3p promoted hMSC homing to the injured liver and resulted in significantly higher engraftment upon peripheral infusion. hMSCs reduced hepatic necrosis and inflammatory infiltration but showed little effect on extracellular matrix (ECM) deposition. By contrast, hMSCs overexpressing miR-9-5p or miR-221-3p resulted in not only less centrilobular necrosis and venous congestion but also a significant reduction of ECM deposition, leading to obvious improvement of hepatocyte morphology and alleviation of fibrosis around central vein and portal triads. Further studies showed that hMSCs inhibited the activation of hepatic stellate cells (HSCs) but could not decrease the expression of TIMP-1 upon acute injury and the expression of MCP-1 and TIMP-1 upon chronic injury, while hMSCs overexpressing miR-9-5p or miR-221-3p led to further inactivation of HSCs and downregulation of all three fibrogenic and proinflammatory factors TGF-β, MCP-1, and TIMP-1 upon both acute and chronic injuries. Overexpression of miR-9-5p or miR-221-3p significantly downregulated the expression of α-SMA and Col-1α1 in activated human hepatic stellate cell line LX-2, suggesting that miR-9-5p and miR-221-3p may partially contribute to the alleviation of liver injury by preventing HSC activation and collagen expression, shedding light on improving the therapeutic efficacy of hMSCs via microRNA modification.

As an economically valuable red seaweed, Neopyropia yezoensis (Rhodophyta) is cultivated in intertidal areas, and its growth and development are greatly influenced by environmental factors such as temperature. Although much effort has been devoted to delineating the influence, the underlying cellular and molecular mechanisms remain elusive. In this study, the gametophyte blades and protoplasts were cultured at different temperatures (13 °C, 17 °C, 21 °C, 25 °C). Only blades cultured at 13 °C maintained a normal growth state (the relative growth rate of thalli was positive, and the content of phycobiliprotein and pigments changed little); the survival and division rates of protoplasts were high at 13 °C, but greatly decreased with the increase in temperature, suggesting that 13 °C is suitable for the growth of N. yezoensis. In our efforts to delineate the underlying mechanism, a partial coding sequence (CDS) of Cyclin B and the complete CDS of cyclin-dependent-kinase B (CDKB) in N. yezoensis were cloned. Since Cyclin B controls G2/M phase transition by activating CDK and regulates the progression of cell division, we then analyzed how Cyclin B expression in the gametophyte blades might change with temperatures by qPCR and Western blotting. The results showed that the expression of Cyclin B first increased and then decreased after transfer from 13 °C to higher temperatures, and the downregulation of Cyclin B was more obvious with the increase in temperature. The phosphorylation of extracellular signal-regulated kinase (ERK) decreased with the increase in temperature, suggesting inactivation of ERK at higher temperatures; inhibition of ERK by FR180204 significantly decreased the survival and division rates of protoplasts cultured at 13 °C. These results suggest that downregulation of Cyclin B and inactivation of ERK might be involved in negatively regulating the survival and division of protoplasts and the growth of gametophyte blades of N. yezoensis at high temperatures.