摘要
Abstract
Objective:To investigate the influence of piperine on the autophagy in oxaliplatin-induced neuropathic pain of mice and its associated mechanisms.Methods:Healthy male C57BL/6 mice aged 6-8 weeks were divided randomly into 6 groups,namely,the control group,the oxaliplatin group(3 mg/kg),three groups of oxaliplatin+piperine(with doses of 10 mg/kg,20 mg/kg and 40 mg/kg respectively),and the piperine group(40 mg/kg),which were all fed for 28 days.The thermal and mechanical allodynia behavior was detected on Day 0,7,14,21 and 28 during the experiment.Then the mice were sacrificed and Dorsal Root Ganglion(DRG)cells were collected for subsequent tests.Primary DRG cells were extracted from neonatal SD mice and divided into 6 groups,including the control group,the oxaliplatin(3μmol/L)group,and four oxaliplatin+piperine groups(with doses of 9μmol/L,12μmol/L,18μmol/L and 24μmol/L respectively).Several tests were conducted after drug treatment for 48 h.The cell survival rate and the expression of autophagy-related genes(including Beclin1,Map1lc3b and SQSTM1)in the tissue and cells of the six groups were de-tected by the CCK-8 method and qRT-PCR,respectively.Western Blot and immunofluorescence staining methods were used to analyze the expression of Beclin1,LC3B and p62 proteins in tissue and cells of the six groups.Results:The ox-aliplatin(3μmol/L)group showed thermal and mechanical allodynia from Day 7(P＜0.05)and symptoms lasted to the end of the test period.The administration of piperine greatly relieved oxaliplatin-induced allodynia(P＜0.05).Moreo-ver,piperine increased the abnormal expression of Beclin1 and Map1lc3a mRNAs and proteins and inhibited the over-expression of p62 proteins caused by oxaliplatin in the DRG tissue of oxaliplatin-induced peripheral neurotoxicity(OIPN)mice(both P＜0.05).Compared with those in the control group,the DRG cell survival rate and the levels of Beclin1 and Map1lc3a mRNAs and proteins were greatly reduced while the levels of SQSTM1 mRNAs and proteins were significantly increased in the oxaliplatin(3μmol/L)group(both P＜0.05).By comparing the oxaliplatin(3μmol/L)group with oxaliplatin+piperine(9μmol/L,12μmol/L,18μmol/L and 24μmol/L)groups,it was found that the use of different concentrations of piperine remarkably improved the survival rate of DRG cells,corrected the significant de-crease in the expression of Beclin1 and Map1lc3b mRNAs(P＜0.05),and inhibited the over-expression of SQSTM1 mRNAs and proteins caused by oxaliplatin(P＜0.05).Conclusion:Piperine has a protective effect on oxaliplatin-in-duced peripheral neuropathy,which is reflected in its ability to increase the survival rate of DRG cells,reduce nerve in-jury,and alleviate hyperalgesia in oxaliplatin-induced neuropathic mice.This protective effect is possibly attributed to the fact that piperine regulates the autophagy process through regulating the expression of Beclin1,LC3B and p62.Pip-erine promotes the expression of Beclin1 and LC3B,while inhibiting the expression of p62,thus mitigating autophagic damage in DRG cells and tissues caused by oxaliplatin,so as to play a protective role in peripheral nerves.关键词
奥沙利铂/背根神经节/胡椒碱/自噬/神经保护Key words
Oxaliplatin/Dorsal Root Ganglion/Piperine/Autophagy/Neuroprotection分类
医药卫生
