Ji Zhu1, Yingling Xu2, Guangyan Ren2, Xiao Hu2, Cui Wang2, Zhen Yang2, Zhuoyu Li2, Wei Mao3, Dezhao Lu4. 1. Clinical Laboratory, The Third Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, China. 2. College of Life Science, Zhejiang Chinese Medical University, Hangzhou, China. 3. Cardiovascular department,The First Affiliated Hospital of Zhejiang Chinese Medicine University, Hangzhou 310006, China. 4. College of Life Science, Zhejiang Chinese Medical University, Hangzhou, China. Electronic address: ludezhao@126.com.
Abstract
BACKGROUND: Tanshinone IIA Sodium sulfonate (STS) is clinically used for treating cardiovascular diseases in Traditional Chinese Medicine due to its antioxidation and anti-inflammation activities. Intracellular chloride channel 1 (CLIC1) participates in the regulation of oxidative stress and inflammation. This study investigates whether CLIC1 mediates the cardioprotective effects of STS. METHODS: STS were used to treat atherosclerosis (AS) induced by feeding Apolipoprotein E-deficient (ApoE-/-) mice with a high-fat, cholesterol-rich diet. In addition, normal and CLIC1-/- human umbilical vein endothelial cells were treated with STS after exposure to H2O2 for 12h. The oxidative status was determined by analyzing reactive oxygen species(ROS) and malondialdehyde (MDA) levels. ELISA, qRT-PCR and Western blot were used to determine the levels of TNF-α, IL-6, ICAM-1 and VCAM-1. CLIC1 cellular localization was examined by immunofluorescence. Chloride ion concentration was detected with chloride ion quenchers (MQAE). RESULTS: STS treatment decreased atherosclerotic lesion area by 3.5 times (P = 0.001) in vivo. Meanwhile, STS reduced MDA production (13.6%, P = 0.008), increased SOD activity (113.6%, P = 0.008), decreased TNF-α (38.6%, P = 0.008) and IL-6 (43.0%, P = 0.03) levels, and downregulated the expression of CLIC1, ICAM-1, and VCAM-1 in the atherosclerotic mice. The dose-dependent anti-oxidative and anti-inflammatory effects of STS were further confirmed in vitro. Furthermore, CLIC1 depletion abolished the STS-mediated decrease of ROS and MDA production in HUVEC cells. Additionally, STS inhibited both CLIC1 membrane translocation and chloride ion concentration. CONCLUSION: The anti-oxidant, and anti-inflammation properties of STS in preventing AS is mediated by its inhibition of CLIC1 expression and membrane translocation.
BACKGROUND: Tanshinone IIA Sodium sulfonate (STS) is clinically used for treating cardiovascular diseases in Traditional Chinese Medicine due to its antioxidation and anti-inflammation activities. Intracellular chloride channel 1 (CLIC1) participates in the regulation of oxidative stress and inflammation. This study investigates whether CLIC1 mediates the cardioprotective effects of STS. METHODS: STS were used to treat atherosclerosis (AS) induced by feeding Apolipoprotein E-deficient (ApoE-/-) mice with a high-fat, cholesterol-rich diet. In addition, normal and CLIC1-/- human umbilical vein endothelial cells were treated with STS after exposure to H2O2 for 12h. The oxidative status was determined by analyzing reactive oxygen species(ROS) and malondialdehyde (MDA) levels. ELISA, qRT-PCR and Western blot were used to determine the levels of TNF-α, IL-6, ICAM-1 and VCAM-1. CLIC1 cellular localization was examined by immunofluorescence. Chloride ion concentration was detected with chloride ion quenchers (MQAE). RESULTS: STS treatment decreased atherosclerotic lesion area by 3.5 times (P = 0.001) in vivo. Meanwhile, STS reduced MDA production (13.6%, P = 0.008), increased SOD activity (113.6%, P = 0.008), decreased TNF-α (38.6%, P = 0.008) and IL-6 (43.0%, P = 0.03) levels, and downregulated the expression of CLIC1, ICAM-1, and VCAM-1 in the atheroscleroticmice. The dose-dependent anti-oxidative and anti-inflammatory effects of STS were further confirmed in vitro. Furthermore, CLIC1 depletion abolished the STS-mediated decrease of ROS and MDA production in HUVEC cells. Additionally, STS inhibited both CLIC1 membrane translocation and chloride ion concentration. CONCLUSION: The anti-oxidant, and anti-inflammation properties of STS in preventing AS is mediated by its inhibition of CLIC1 expression and membrane translocation.