This study aimed to investigate the role of tanshinone IIA (TSO IIA) in astrocytic swelling caused by ischemia–reperfusion-like injury in an in vitro model and the molecular mechanisms underlying this effect. Primary brain astrocytes were cultured under conditions of glucose and oxygen deprivation and reoxygenation (OGD/R). The study explored the effects of TSO IIA treatment on cell swelling and injury and the protein levels of aquaporin 4 (AQP4) in the plasma membrane. It then examined the involvement of the high-mobility group box protein 1 (HMGB1)/receptors for advanced-glycation end products (RAGE)/nuclear factor-kappa B (NF-κB)/interleukin-6 (IL-6) pro-inflammatory axis in TSO IIA-mediated protection. The treatment with TSO IIA alleviated OGD/R-induced astrocytic swelling and the overclustering of AQP4 protein in the plasma membrane. In addition, TSO IIA significantly reduced the overexpression of HMGB1 and the high levels of the NF-κB protein in the nucleus and of the IL-6 protein in the cytoplasm and extracellular media induced by OGD/R. The combination of TSO IIA and recombinant HMGB1 reversed these effects. The inhibition of the RAGE, the receptor of HMGB1, induced results similar to those of TSO IIA. In addition, exogenous IL-6 reversed TSO IIA-mediated effect on AQP4 overclustering and cell swelling. TSO IIA significantly reduced astrocyte swelling after OGD/R injury in vitro, via blocking the activation of the HMGB1/RAGE/NF-κB/IL-6 pro-inflammatory axis and thereby decreasing the expression of AQP4 in the plasma membrane.

Here, we investigated the role of tanshinone IIA (TSO IIA) in astrocytic swelling caused by ischemia and reperfusion-like injury in an in vitro model and the molecular mechanisms underlying this effect. Primary brain astrocytes were cultured in conditions of glucose and oxygen deprivation and reoxygenation (OGD/R). We investigated the effects of TSO IIA treatment on cell swelling and injury, and the expression levels of AQP4 protein in the plasma membrane. We then studied the involvement of the HMGB1/RAGE/NF-κB/IL-6 pro-inflammatory axis in TSO IIA-mediated protection. Treatment with TSO IIA alleviated OGD/R-induced astrocytic swelling and the over-clustering of AQP4 protein in the plasma membrane. In addition, TSO-IIA significantly reduced over-expression of HMGB1, the high levels of NF-κB protein in the nucleus, and of IL-6 protein in cytoplasm and extracellular media induced by OGD/R. Combination of TSO IIA and recombinant HMGB1 reversed these effects. Inhibition of RAGE, the receptor of HMGB1, induced similar results to those of TSO IIA. In addition, exogenous IL-6 reversed TSO IIA-mediated effect on AQP4 over-clustering and cell swelling.TSO IIA significantly reduced astrocyte swelling after OGD/R injury in vitro, via blocking activation of the HMGB1/RAGE/NF-κB/IL-6 pro-inflammatory axis and thereby decreasing the expression of AQP4 in plasma membrane.

Background: Tanshione IIA (TSO IIA) has emerged as a powerful anti-inflammatory compound with multiple therapeutic properties. Here, we investigated the role of TSO IIA in astrocytic swelling caused by ischemia and reperfusion-like injury in an in vitro model and the molecular mechanisms underlying this effect. Methods: Primary brain astrocytes were cultured in vitro in conditions of glucose and oxygen deprivation and reoxygenation (OGD/R). We investigated the effects of TSO IIA treatment on cell swelling and injury, and the expression levels of AQP4 protein in the plasma membrane. We then studied the involvement of the HMGB1/RAGE/NF-κB/IL-6 pro-inflammatory axis in TSO IIA-mediated protection. Results: Treatment with TSO IIA alleviated OGD/R-induced astrocytic swelling, changes in cell morphology and ultrastructure, the release of LDH, and the over-clustering of AQP4 protein in the plasma membrane. In addition, TSO-IIA significantly reduced over-expression of HMGB1 protein in the cytoplasm and surrounding medium, the high levels of NF-κB protein in the nucleus, and of IL-6 protein in cytoplasm and extracellular media induced by OGD/R. Combination of TSO IIA and recombinant HMGB1 (rHMGB1) reversed these effects. Inhibition of RAGE, the receptor of HMGB1, via administration of FPS-ZM1 (a RAGE antagonist) induced similar results to those of TSO IIA. In addition, exogenous IL-6 reversed TSO IIA-mediated effect on AQP4 protein clustering in the plasma membrane, cell swelling and LDH activity.Conclusions: TSO IIA significantly reduced astrocyte swelling after OGD/R injury in vitro, via blocking activation of the HMGB1/RAGE/NF-κB/IL-6 pro-inflammatory axis and thereby decreasing the expression of AQP4 in plasma membrane.

Background: Tanshione IIA (TSO IIA) has emerged as a powerful anti-inflammatory compound with multiple therapeutic properties. Here, we investigated the role of TSO IIA in astrocytic swelling caused by ischemia and reperfusion-like injury in an in vitro model and the molecular mechanisms underlying this effect. Methods: Primary brain astrocytes were cultured in vitro in conditions of glucose and oxygen deprivation and reoxygenation (OGD/R). We investigated the effects of TSO IIA treatment on cell swelling and injury, and the expression levels of AQP4 protein in the plasma membrane. We then studied the involvement of the HMGB1/RAGE/NF-κB/IL-6 pro-inflammatory axis in TSO IIA-mediated protection. Results: Treatment with TSO IIA alleviated OGD/R-induced astrocytic swelling, changes in cell morphology and ultrastructure, the release of LDH, and the over-clustering of AQP4 protein in the plasma membrane. In addition, TSO-IIA significantly reduced over-expression of HMGB1 protein in the cytoplasm and surrounding medium, the high levels of NF-κB protein in the nucleus, and of IL-6 protein in cytoplasm and extracellular media induced by OGD/R. Combination of TSO IIA and recombinant HMGB1 (rHMGB1) reversed these effects. Inhibition of RAGE, the receptor of HMGB1, via administration of FPS-ZM1 (a RAGE antagonist) induced similar results to those of TSO IIA. The combination of HMGB1 and FPS-ZM1 did not alter the effects mediated by FPS-ZM1 alone. In addition, exogenous IL-6 reversed TSO IIA-mediated effect on AQP4 protein clustering in the plasma membrane, cell swelling and LDH activity.Conclusions: TSO IIA significantly reduced astrocyte swelling after OGD/R injury in vitro, via blocking activation of the HMGB1/RAGE/NF-κB/IL-6 pro-inflammatory axis and thereby decreasing the expression of AQP4 in plasma membrane.