Wang X.,Key Laboratory of Xinan Medicine |
Wang X.,Anhui University of Traditional Chinese Medicine |
Zhu G.,Key Laboratory of Xinan Medicine |
Zhu G.,Anhui University of Traditional Chinese Medicine
Zhongguo Zhongyao Zazhi | Year: 2012
Objective: To study the protective effect and mechanism of salvianolic acid B (Sal B) on glutamate-induced excitotoxicity. Method: Glutamate-induced PC12 cell injury model was established to detect the cell survival rate by MTT, the leakage rate of lactic dehydrogenases using LDH, and the cell apoptosis by using AO/EB double staining for fluorescence microscope and PI single staining flow cytometry which was also used to detect the content of intracellular reactive oxygen species. The expression of Caspase-3 protein was also detected by the Western blotting method. Result: Sal B is proved to inhibit glutamate-induced PC12 cells from injury and prevent them from releasing LDH within the range from 50 μmol·L-1 to 200 μmol·L-1. Meanwhile, Sal B has an effect on significantly reducing the expression of inhibit glutamate-induced active Caspase-3 protein, inhibiting accumulated glutamate-induced ROS and decreasing PC12 cell apoptosis rate within the range from 50 μmol·L-1 to 200 μmol·L-1. Conclusion: The study proves that Sal B prevented against glutamate-induced cell injury via inhibiting ROS formation and Caspase-3 pathway-dependent apoptosis in PC12 cells.
Liu Y.-R.,Key Laboratory of Chinese Medicine Research and Development |
Chen J.-J.,Key Laboratory of Xinan Medicine |
Dai M.,Anhui University of Traditional Chinese Medicine
Acta Pharmacologica Sinica | Year: 2014
Aim: Paeonol (2'-hydroxy-4'-methoxyacetophenone) from Cortex moutan root is a potential therapeutic agent for atherosclerosis. This study sought to investigate the mechanisms underlying anti-inflammatory effects of paeonol in rat vascular endothelial cells (VECs) in vitro.Methods:VECs were isolated from rat thoracic aortas. The cells were pretreated with paeonol for 24 h, and then stimulated with ox-LDL for another 24 h. The expression of microRNA-21 (miR-21) and PTEN in VECs was analyzed using qRT-PCR. The expression of PTEN protein was detected by Western blotting. TNF-α release by VECs was measured by ELISA. Results: Ox-LDL treatment inhibited VEC growth in dose- and time-dependent manners (the value of IC 50 was about 20 mg/L at 24 h). Furthermore, ox-LDL (20 mg/L) significantly increased miR-21 expression and inhibited the expression of PTEN, one of downstream target genes of miR-21 in VECs. In addition, ox-LDL (20 mg/L) significantly increased the release of TNF-α from VECs. Pretreatment with paeonol increased the survival rate of ox-LDL-treated VECs in dose- and time-dependent manners. Moreover, paeonol (120 μmol/L) prevented ox-LDL-induced increases in miR-21 expression and TNF-α release, and ox-LDL-induced inhibition in PTEN expression. A dual-luciferase reporter assay showed that miR-21 bound directly to PTEN's 3'-UTR, thus inhibiting PTEN expression. In ox-LDL treated VECs, transfection with a miR-21 mimic significantly increased miR-21 expression and inhibited PTEN expression, and attenuated the protective effects of paeonol pretreatment, whereas transfection with an miR-21 inhibitor significantly decreased miR-21 expression and increased PTEN expression, thus enhanced the protective effects of paeonol pretreatment. Conclusion: miR-21 is an important target of paeonol for its protective effects against ox-LDL-induced VEC injury, which may play critical roles in development of atherosclerosis. © 2014 CPS and SIMM.
Yang Y.,Anhui University |
Yang Y.,Key Laboratory of Xinan Medicine |
Wang F.,Anhui University |
Yin D.,Anhui University |
And 5 more authors.
Colloids and Surfaces B: Biointerfaces | Year: 2015
Tissue engineering scaffolds (TES) can carry numerous biomacromolecules and cells, and they have been widely used in diabetic skin wound healing with positive effects. However, the bioactive retention of biomacromolecules and cells during fabrication and storage is still a factor restricting their use. Moreover, impaired blood supply in/around poorly healing diabetic skin wounds has not been considered. In the present study, a bioactive natural substance of Astragalus polysaccharide (APS), which has stable and confirmed effects on endothelial protection, was embedded into fibrous TES by electrospinning. The administration of APS-loaded TES on the skin wound in a diabetic rat model led to a dose-dependent promotion in skin blood flow around wounds and an increase in endoglin expression and microvessel density in regenerated skin tissues. Furthermore, the higher loading of APS in TES led to faster collagen synthesis, appendage and epidermal differentiation, and wound closure. In summary, the combination of APS with TES is a potentially novel therapeutic strategy for diabetic skin wound healing, as it not only mimics the ultrastructure of extracellular matrixes but also restores skin microcirculation. © 2015 Elsevier B.V..
Yang G.,Key Laboratory of Xinan Medicine |
Yang G.,Anhui University |
Yang G.,Anhui Academy of Chinese Medicine |
Qian C.,Key Laboratory of Xinan Medicine |
And 17 more authors.
Cellular and Molecular Neurobiology | Year: 2016
Tetramethylpyrazine (TMP, also known as Ligustrazine), which is isolated from Chinese Herb Medicine Ligustium wollichii Franchat (Chuan Xiong), has been widely used in China for the treatment of ischemic stroke by Chinese herbalists. Brain microvascular endothelial cells (BMECs) are the integral parts of the blood–brain barrier (BBB), protecting BMECs against oxygen-glucose deprivation (OGD) which is important for the treatment of ischemic stroke. Here, we investigated the protective mechanisms of TMP, focusing on OGD-injured BMECs and the Rho/Rho-kinase (Rho-associated kinases, ROCK) signaling pathway. The model of OGD-injured BMECs was established in this study. BMECs were identified by von Willebrand factor III staining and exposed to fasudil, or TMP at different concentrations (14.3, 28.6, 57.3 µM) for 2 h before 24 h of OGD injury. The effect of each treatment was examined by cell viability assays, measurement of intracellular reactive oxygen species (ROS), and transendothelial electric resistance and western blot analysis (caspase-3, endothelial nitric oxide synthase (eNOS), RhoA, Rac1). Our results show that TMP significantly attenuated apoptosis and the permeability of BMECs induced by OGD. In addition, TMP could notably down-regulate the characteristic proteins in Rho/ROCK signaling pathway such as RhoA and Rac1, which triggered abnormal changes of eNOS and ROS, respectively. Altogether, our results show that TMP has a strong protective effect against OGD-induced BMECs injury and suggest that the mechanism might be related to the inhibition of the Rho/ROCK signaling pathway. © 2016 Springer Science+Business Media New York