Brain derived neurotrophic factor alleviates neuronal injury induced by ropivacaine through threonine protein kinase signaling pathway
10.3760/cma.j.issn.1671-8925.2020.02.008
- VernacularTitle:BDNF通过Akt信号通路减轻罗哌卡因诱导的神经细胞损伤
- Author:
Yulin ZHU
1
;
Jingying LIU
;
Xiangyong LIU
Author Information
1. 烟台山医院麻醉科
- Keywords:
Brain derived neurotrophic factor;
Ropivacaine;
Protein kinase B;
Neuron
- From:
Chinese Journal of Neuromedicine
2020;19(2):154-163
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the protective effect of brain-derived neurotrophic factor (BDNF) on neuronal injury induced by ropivacaine (Rop) and its mechanism.Methods (1) Experiment one:0,1,2,3,4 and 5 mmol/L Rop was used to stimulate SH-SY5Y cells for 48 h to induce neuronal injury;the morphological changes of the cells were observed under microscope;MTT assay was used to detect the cell activity;flow cytometry was used to detect the cell apoptosis,and immunohistochemistry was used to detect the expressions of protein kinase B (Akt) and proliferating cell nuclear antigen (PCNA).(2) Experiment two:SH-SY5Y cells were treated with 0,1,3,5,7 mmol/L Rop,respectively;the cell activity was measured 48 h after Rop treatment;the semi inhibitory concentration (IC50) of Rop was calculated by MTT assay;the SH-SY5Y cells were divided into control group (PBS for 48 h),Rop group (Rop at IC50 for 48 h),BDNF+Rop group (20 μg/L BDNF for 2 h,and Rop at IC50 for 48 h),Akt pathway activator SC79+Rop group (5 mg/L SC79 for 2 h,and Rop at IC50 for 48 h),and BDNF+Akt pathway inhibitor API-2+Rop group (20 μg/L BDNF+10 μmol/L API-2 for 2 h,Rop at IC50 for 48 h);the morphological changes of the cells were observed under microscope;MTT assay was used to detect the cell activity;flow cytometry was used to detect the cell apoptosis,and immunohistochemistry was used to detect the expressions of Akt and PCNA;the expressions of B lymphoma-2 (Bcl-2),Bcl-2-associated X (Bax) and cysteine protease-3 (Caspase-3) were detected by reverse transcription (RT)-PCR and Western blotting.Western blotting was used to detect the expressions of Akt and phosphatidylinositol 3-kinase (PI3K).Results (1) As compared with the 0 mmol/L Rop group,the 1,2,3,4,and 5 mmol/L Rop group had significantly decreased cell activity,significantly increased apoptosis rate,and statistically smaller number of Akt and PCNA positive cells (P＜0.05).(2) As compared with the control group,the Rop group had significantly decreased cell activity,statistically increased apoptosis rate,significantly smaller number of Akt and PCNA positive cells,significantly decreased Bcl-2 mRNA and protein expressions,significantly increased Bax and caspase-3 mRNA and protein expressions,and significantly decreased phosphorylated-(p-) Akt and p-PI3K protein expressions;as compared with the Rop group,the BDNF+Rop group and SC79+Rop group had significantly higher cell activity,significantly decreased apoptosis rate,significantly larger number of Akt and PCNA positive cells,significantly increased Bcl-2 mRNA and protein expressions,statistically decreased mRNA and protein expressions of Bax and Caspase-3,and significantly increased p-Akt and p-PI3K protein expressions (P＜0.05);as compared with the BDNF+Rop group and SC79+Rop group,the BDNF+API-2+Rop group had significantly lower cell activity,significantly increased apoptosis rate,significantly smaller number of Akt and PCNA positive ceils,significantly decreased Bcl-2 mRNA and protein expressions,statistically increased mRNA and protein expressions of Bax and Caspase-3,and significantly decreased p-Akt and p-PI3K protein expressions (P＜0.05).Conclusion BDNF can alleviate ropivacaine-induced neuronal injury by activating Akt signaling pathway,consequently modulating the proliferation and apoptosis of neurons.
