title: Andrographolide sodium bisulfate-induced apoptosis and autophagy in human proximal tubular endothelial cells is a ROS-mediated pathway.
abstract: OBJECTIVE
  The nephrotoxic mechanisms of andrographolide sodium bisulfate (ASB) remain largely unknown. This study attempted to explore the mechanism of ASB-induced nephrotoxicity using human proximal tubular endothelial cells (HK-2).
  
  
  METHODS
  For this study HK-2 cells were treated with rising concentrations of ASB. Their survival rate was detected using MTT assay and ultrastructure was observed with electron microscopy. L-Lactate dehydrogenase (LDH) assay was followed by examination of mitochondrial membrane potential (MMP). Reactive oxygen species (ROS) was detected using different methods and apoptosis/autophage related proteins were detected using immunoblotting.
  
  
  RESULTS
  We found that ASB inhibited HK-2 cell proliferation and decreased cell survival rate in a time and dose-dependent manner (P<0.05, P<0.01, respectively). With increasing ASB concentration, cell structure was variably damaged and evidence of apoptosis and autophagy were observed. MMP gradually decreased and ROS was induced. The expression of JNK and Beclin-1 increased and activation of the JNK signaling pathway were seen. Apoptosis was induced via the mitochondrial-dependent caspase-3 and caspase-9 pathway, and autophagy related protein Beclin-1 was enhanced by ASB.
  
  
  CONCLUSIONS
  The data show that ASB induces high levels of ROS generation in HK-2 cells and activates JNK signaling. Furthermore, ASB induces cell apoptosis via the caspase-dependent mitochondrial pathway, and induces cellular autophagy, in part by enhancing Beclin-1 protein expression.
authors.affiliation: School of Pharmacology, Zhejiang Chinese Medical University, Hangzhou 310053, Zhejiang Province, China. Electronic address: 2698669983@qq.com.
authors.forename: Hong
authors.initials: H
authors.lastname: Lu
authors.affiliation: Institute of Materia Medica, Zhejiang Academy of Medical Sciences, Hangzhou 310013, Zhejiang Province, China.
authors.forename: Xin-Yue
authors.initials: XY
authors.lastname: Zhang
authors.affiliation: School of Pharmacology, Zhejiang Chinese Medical University, Hangzhou 310053, Zhejiang Province, China.
authors.forename: Yi-Qi
authors.initials: YQ
authors.lastname: Wang
authors.affiliation: Institute of Materia Medica, Zhejiang Academy of Medical Sciences, Hangzhou 310013, Zhejiang Province, China.
authors.forename: Xiao-Liang
authors.initials: XL
authors.lastname: Zheng
authors.affiliation: Institute of Materia Medica, Zhejiang Academy of Medical Sciences, Hangzhou 310013, Zhejiang Province, China.
authors.lastname: Yin-Zhao
authors.affiliation: School of Pharmacology, Zhejiang Chinese Medical University, Hangzhou 310053, Zhejiang Province, China.
authors.forename: Wen-Min
authors.initials: WM
authors.lastname: Xing
authors.affiliation: School of Pharmacology, Zhejiang Chinese Medical University, Hangzhou 310053, Zhejiang Province, China.
authors.forename: Qin
authors.initials: Q
authors.lastname: Zhang
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D001343:Autophagy / Q000187:drug effects; D000071186:Beclin-1; D053148:Caspase 3 / Q000378:metabolism; D053453:Caspase 9 / Q000378:metabolism; D002460:Cell Line; D045304:Cytochromes c / Q000378:metabolism; D004224:Diterpenes / Q000633:toxicity*; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism*; D007687:Kidney Tubules, Proximal / Q000166:cytology; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008565:Membrane Proteins / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013431:Sulfates / Q000633:toxicity*; D013482:Superoxide Dismutase / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D051017:Apoptosis Regulatory Proteins; C494826:BAX protein, human; C491997:BECN1 protein, human; D000071186:Beclin-1; D004224:Diterpenes; D008565:Membrane Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D017382:Reactive Oxygen Species; D013431:Sulfates; D051028:bcl-2-Associated X Protein; C012036:sodium sulfate; C030419:andrographolide; D045304:Cytochromes c; D013482:Superoxide Dismutase; D048031:JNK Mitogen-Activated Protein Kinases; D053148:Caspase 3; D053453:Caspase 9; D005978:Glutathione
keywords: Andrographolide sodium bisulfate; Apoptosis; Autophagy; Human proximal tubular endothelial cell; Reactive oxygen species