摘要
目的探讨不同浓度的丙泊酚预处理对SK-N-SH神经母细胞缺氧/复氧损伤的保护作用及对神经生长因子(NGF)表达的影响。方法取离体培养的SK-N-SH细胞,随机分成7组:对照组,不作任何处理;氯化钴(CoCl2)组:加入300μmol/L CoCl2处理1 h,然后更换正常的培养基培养24 h,之后更换无血清的培养基培养;脂肪乳剂组:加入10%脂肪乳剂90μL预处理1 h后加入300μmol/L CoCl2;丙泊酚组:培养孔中加入10、20、50、100μmol/L浓度的丙泊酚预处理1 h后,同CoCl2组处理。MTT法测定细胞增殖,流式细胞技术测定细胞的凋亡。为进一步研究不同浓度丙泊酚对NGF表达的影响,将SK-N-SH细胞分为6组,分别为对照组、CoCl2组和10、20、50、100μmol/L浓度的丙泊酚,用RT-PCR和Western blot分别检测NGF mRNA和NGF蛋白表达。为探讨丙泊酚对NGF的作用机制,将细胞随机分为6组:对照组、CoCl2组、50μmol/L丙泊酚组、10μmol/L PD98059组、30μmol/L SP600125组和30μmol/L SB202190组。结果脂肪乳剂组细胞活性与CoCl2组比较,差异无统计学意义(P>0.05),50μmol/L丙泊酚预处理可以明显增加SK-N-SH细胞的增殖能力,减少凋亡(P<0.01)。10、20、100μmol/L浓度的丙泊酚与CoCl2组比较差异无统计学意义(P>0.05)。50μmol/L丙泊酚预处理上调NGF mRNA和NGF蛋白的表达,而这些调节作用可被JNK抑制剂SP600125抑制(P<0.01),PD98059和SB202190对NGF的表达与CoCl2组比较,差异无统计学意义(P>0.05)。结论 50μmol/L丙泊酚预处理对缺氧/复氧后的SK-N-SH细胞有保护作用,NGF在丙泊酚的预处理中起到重要的作用。
Objective To establish a model of hypoxia/re - oxygenation (H/R) injury, as to examine the neuro- protective effect of propofol, and to investigate the role of nerve growth factor (NGF) in it. Methods SK - N - SH cells subjected to I-I/R were pretreated with different concentrations of propofol, after which viability and apoptosis of the cells were determined by MIT assay and Annexin V flow cytometry, respectively. Meanwhile, the expression of NGF was meas- ured by reverse transcriptase polymerase chain reaction and Western blot. Results H/R resulted in reduced cell viability and increased cell apoptosis in SK -N -SH cells, as indicated by MTT assay and Annexin V flow cytometry, respectively. Pretreatment with 50μmol/L propofol reversed I-I/R - induced neurotoxicity and induced a remarkable increase in NGF mRNA expression and the protein expression. C -Jun N -terminal kinase (JNK) suppressed the effect of propofol on the NGF expression and altered its neuroprotective effec. Conclusion These findings suggest the potential of propofol for protecting SK - N - SH cells against H/R by increasing endogenous neurotrophin generation through a JNK signaling pathway.
出处
《广东医学》
CAS
CSCD
北大核心
2013年第22期3379-3381,共3页
Guangdong Medical Journal
基金
辽宁省科学技术计划项目(编号:2011225020)