摘要
目的:探讨马尾松松针提取物(PNE)对大鼠脑缺血再灌注后氧化应激损伤的保护作用。方法:将SD雄性大鼠随机分为手术对照组、模型对照组、依达拉奉组、PNE小剂量组(200 mg/kg)、PNE中剂量组(400 mg/kg)、PNE大剂量组(800 mg/kg),于造模前连续7 d及造模后6 h分别灌胃给药。其中,手术对照组和模型对照组灌胃给予等渗氯化钠溶液,依达拉奉组灌胃给予等渗氯化钠溶液的同时腹腔注射依达拉奉3 mg/kg,PNE各组灌胃给予各剂量PNE。通过大脑中动脉阻塞法建立脑缺血再灌注损伤大鼠模型,于再灌注24 h后测定各组大鼠神经功能缺损评分、脑组织含水量、脑梗死体积。采用苏木素-伊红染色观察大脑皮层及海马组织结构病理变化并统计正常神经细胞数;TUNEL法检测大脑皮层神经细胞凋亡率;试剂盒检测缺血侧脑组织一氧化氮、丙二醛含量与超氧化物歧化酶(SOD)活性;蛋白质印迹法检测大脑皮层c-Jun氨基末端激酶(JNK)3、磷酸化JNK3、B淋巴细胞瘤蛋白(Bcl)-2、Bcl-2相关X蛋白(Bax)、细胞色素C、胱天蛋白酶(caspase)-3的蛋白表达水平。结果:与模型对照组比较,PNE各剂量组行为学评分、脑组织含水量及脑梗死体积均显著降低(均P<0.05),大脑皮层及海马CA1区组织病理性损伤显著减轻,缺血侧皮层及海马CA1区中正常神经细胞数增加(均P<0.05),以PNE中剂量组效果最好。与模型对照组比较,PNE中剂量组皮层神经细胞凋亡率显著减小,大脑皮层组织中一氧化氮、丙二醛含量显著减少,SOD活性显著升高,磷酸化JNK3/JNK3比值、细胞色素C、caspase-3蛋白表达水平均显著降低,Bcl-2/Bax比值显著升高(均P<0.05)。结论:PNE对脑缺血再灌注损伤具有保护作用,这种保护作用可能与清除一氧化氮和丙二醛,抑制氧化应激介导的JNK3/caspase-3信号转导来抑制神经细胞凋亡有关。
Objective:To investigate the effect and mechanism of Pinus massoniana needle extracts(PNE)on oxidative stress injury in cerebral ischemia reperfusion rats.Methods:The SD male rats were randomly divided into sham group,model control group,Edaravone(3 mg/kg)group,PNE low-dose(200 mg/kg),medium-dose(400 mg/kg)and high-dose(800 mg/kg)groups.PNE was administered by gavage for 7 d before modeling and 6 h after modeling in PNE treatment groups;Edaravone was given by intraperitoneal injection 7 d before modeling and 6 h after reperfusion.The rat model of cerebral ischemia reperfusion injury was established by middle cerebral artery occlusion method.After 24 h of reperfusion,the neurological deficit score,brain water content and cerebral infarction volume of rats were measured.The pathological changes of cerebral cortex and hippocampus were observed by HE staining,and the number of normal nerve cells was counted.The apoptosis rate of neurons in cerebral cortex was detected by TUNEL method.The content of nitric oxide(NO),malondialdehyde(MDA)and superoxide dismutase(SOD)activity in ischemic brain tissue were detected.The protein expression of c-Jun N-terminal kinase(JNK)3,phosphorylated JNK3(p-JNK3),B-cell lymphoma protein(Bcl)-2,Bcl-2 associated X(Bax),cytochrome C and caspase-3 in cerebral cortex were detected by Western blotting method.Results:Compared with the model control group,the behavioral score,brain water content and cerebral infarction volume in PNE groups were significantly reduced(all P<0.05),the pathological damage of cerebral cortex and hippocampal CA1 area was significantly alleviated,and the number of normal nerve cells in ischemic cortex and hippocampal CA1 area was increased(all P<0.05).The medium-dose PNE group had the best effect.Compared with the model control group,the apoptosis rate of cortical neurons,the content of NO and MDA in cerebral cortex,the ratio of p-JNK3/JNK3,the expression level of cytochrome C and caspase-3 protein in PNE medium-dose group were significantly reduced,and the activity of
作者
章蕾
王凌峰
胡培豪
钟晓明
张丹妮
仇凤梅
黄真
ZHANG Lei;WANG Lingfeng;HU Peihao;ZHONG Xiaoming;ZHANG Danni;QIU Fengmei;HUANG Zhen(School of Pharmaceutical Sciences,Zhejiang Chinese Medical University,Hangzhou 310053,China;Zhejiang Provincial Key Laboratory of Neuropharmacology and Translational Medicine,Hangzhou 310053,China)
出处
《浙江大学学报（医学版）》
CAS
CSCD
北大核心
2022年第5期563-572,共10页
Journal of Zhejiang University(Medical Sciences)
基金
国家自然科学基金(81573643)
浙江省自然科学基金(LY20H280010,LY22H280010)
浙江中医药大学校级科研项目(2021ZY14)。
