Icariin Ameliorates D-galactose-induced Cell Injury in Neuron-like PC12 Cells by Inhibiting MPTP Opening  

作　　者：Shan-shan HU Tong-yao WANG Lu NI Fan-xin HU Bo-wen YUE Ying ZHENG Tian-lun WANG Abhishek KUMAR Yan-yan WANG Jin-e WANG Zhi-yong ZHOU 

机构地区：[1]Third-grade Pharmacological Laboratory of Traditional Chinese Medicine,State Administration of Traditional Chinese Medicine,China Three Gorges University,Yichang 443002,China [2]College of Medicine and Health Sciences,China Three Gorges University,Yichang 443002,China [3]College of Basic Medical Science,China Three Gorges University,Yichang 443002,China

出　　处：《Current Medical Science》2024年第4期748-758,共11页当代医学科学（英文）

基　　金：supported by the Natural Science Foundation of Yichang City of China(No.A23-1-075).

摘　　要：Objective Icariin(ICA)has a good neuroprotective effect and can upregulate neuronal basal autophagy in naturally aging rats.Mitochondrial dysfunction is associated with brain aging-related neurodegenerative diseases.Abnormal opening of the mitochondrial permeability transition pore(mPTP)is a crucial factor in mitochondrial dysfunction and is associated with excessive autophagy.This study aimed to explore that ICA protects against neuronal injury by blocking the mPTP opening and down-regulating autophagy levels in a D-galactose(D-gal)-induced cell injury model.Methods A cell model of neuronal injury was established in rat pheochromocytoma cells(PC12 cells)treated with 200 mmol/L D-gal for 48 h.In this cell model,PC12 cells were pre-treated with different concentrations of ICA for 24 h.MTT was used to detect cell viability.Senescence associatedβ-galactosidase(SA-β-Gal)staining was used to observe cell senescence.Western blot analysis was performed to detect the expression levels of a senescence-related protein(p21),autophagy markers(LC3B,p62,Atg7,Atg5 and Beclin 1),mitochondrial fission and fusion-related proteins(Drp1,Mfn2 and Opa1),and mitophagy markers(Pink1 and Parkin).The changes of autophagic flow were detected by using mRFP-GFP-LC3 adenovirus.The intracellular ultrastructure was observed by transmission electron microscopy.Immunofluorescence was used to detect mPTP,mitochondrial membrane potential(MMP),mitochondrial reactive oxygen species(mtROS)and ROS levels.ROS and apoptosis levels were detected by flow cytometry.Results D-gal treatment significantly decreased the viability of PC12 cells,and markedly increased the SA-β-Gal positive cells as compared to the control group.With the D-gal stimulation,the expression of p21 was significantly up-regulated.Furthermore,D-gal stimulation resulted in an elevated LC3B II/I ratio and decreased p62 expression.Meanwhile,autophagosomes and autolysosomes were significantly increased,indicating abnormal activation of autophagy levels.In addition,in this D-gal-induced mo

关 键 词：ICARIIN neuronal injury mitochondrial dysfunction mitochondrial permeability transition pore AUTOPHAGY 

分 类 号：R285[医药卫生—中药学]