title: Protective effects of aloin on oxygen and glucose deprivation-induced injury in PC12 cells.
abstract: The present study aims to determine whether aloin could protect cells from ischemic and reperfusion injury in vitro and to elucidate the related mechanisms. Oxygen and glucose deprivation model in PC12 cells was used in the present study. 2-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), lactate dehydrogenase (LDH) assay and Hoechst 33342 nuclear staining were used to evaluate the protective effects of aloin, at concentrations of 10, 20, or 40 μg/mL in PC12 cells. PCR was applied to detect fluorescence caspase-3, Bax and Bcl-2 mRNA expression in PC12 cells. The contents of malondialdehyde (MDA), superoxide dismutase (SOD) activity were evaluated by biochemical method. The concentration of intracellular-free calcium [Ca(2+)]i, mitochondrial membrane potential (MMP) were determined to estimate the degree of neuronal damage. It was shown that aloin (10, 20, and 40 μg/mL) significantly attenuated PC12 cells damage with characteristics of an increased injured cells absorbance of MTT and releases of LDH, decreasing cell apoptosis, and antagonizing decreases in SOD activity and increase in MDA level induced by OGD-reoxygenation. Meanwhile pretreatment with aloin significantly reduced injury-induced intracellular ROS, increased MMP (P<0.01), but it inhibited [Ca(2+)]i (P<0.01) elevation in a dose-dependent manner. Furthermore, pre-treatment with aloin significantly up-regulated Bcl-2 mRNA expression, down-regulated Bax mRNA expression and consequently activated caspase-3 mRNA expression in a dose-dependent manner. The results indicated that the protection of aloin on OGD-induced apoptosis in PC12 cells is associated with its suppression on OGD-induced oxidative stress and protection on mitochondrial function and inhibition of caspase activity. Alion could be a promising candidate in the development of a novel class of anti-ischemic agent.
authors.affiliation: Department of Pharmacology, College of Pharmacy, Ningxia Medical University, Yinchuan, China.
authors.forename: Renyuan
authors.initials: R
authors.lastname: Chang
authors.affiliation: Department of Pharmacology, College of Pharmacy, Ningxia Medical University, Yinchuan, China.
authors.forename: Ru
authors.initials: R
authors.lastname: Zhou
authors.affiliation: Department of Pharmacology, College of Pharmacy, Ningxia Medical University, Yinchuan, China.
authors.forename: Xue
authors.initials: X
authors.lastname: Qi
authors.affiliation: Department of Pharmacology, College of Pharmacy, Ningxia Medical University, Yinchuan, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Pharmacology, College of Pharmacy, Ningxia Medical University, Yinchuan, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Wu
authors.affiliation: Department of Pharmacology, College of Pharmacy, Ningxia Medical University, Yinchuan, China.
authors.forename: Wenli
authors.initials: W
authors.lastname: Yang
authors.affiliation: Department of Pharmacology, College of Pharmacy, Ningxia Medical University, Yinchuan, China.
authors.forename: Wannian
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Ningxia Key Lab of Craniocerebral Diseases of Ningxia Hui Autonomous Region, Yinchuan, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Sun
authors.affiliation: College of Nursing, Ningxia Medical University, Yinchuan, China. Electronic address: li_yuxiang@163.com.
authors.forename: Yuxiang
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Pharmacology, College of Pharmacy, Ningxia Medical University, Yinchuan, China; Ningxia Hui Medicine Modern Engineering Research Center, Ningxia Medical University, Yinchuan, China. Electronic address: yujq910315@163.com.
authors.forename: Jianqiang
authors.initials: J
authors.lastname: Yu
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D053148:Caspase 3; D015687:Cell Hypoxia / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D004642:Emodin / Q000031:analogs & derivatives* / Q000494:pharmacology; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000187:drug effects*; D005947:Glucose / Q000235:genetics*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D018696:Neuroprotective Agents / Q000494:pharmacology*; D018384:Oxidative Stress / Q000187:drug effects*; D010087:Oxides / Q000378:metabolism; D016716:PC12 Cells; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D017670:Sodium Compounds / Q000378:metabolism; D045162:Thiazolidinediones / Q000378:metabolism; D013997:Time Factors; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: C575035:(Z)2-(5-(4-methoxybenzylidene)-2,4-dioxothiazolidin-3-yl)acetic acid; D018696:Neuroprotective Agents; D010087:Oxides; D019253:Proto-Oncogene Proteins c-bcl-2; D017382:Reactive Oxygen Species; D017670:Sodium Compounds; D045162:Thiazolidinediones; D051028:bcl-2-Associated X Protein; C096707:sodium oxide; D007770:L-Lactate Dehydrogenase; D053148:Caspase 3; D005947:Glucose; D004642:Emodin; D002118:Calcium; C006457:alloin
keywords: Aloin; Apoptosis; Ischemic injury; Oxidative stress; Oxygen and glucose deprivation; PC12 cells