罗汉果苷V调节Shh/Gli1信号通路对急性脑梗死模型大鼠神经损伤的影响
Effect of mogroside V on nerve injury in rats with acute cerebral infarction by regulating Shh/Gli1 signaling pathway
李梅芳 1姜磊1
作者信息
摘要
目的 探究罗汉果苷V(Mogroside V,MogV)调节音猬因子/家族锌指蛋白1(Sonic hedgehog/glioma-associated oncogene 1,Shh/Gli1)信号通路对急性脑梗死(Acute cerebral infarction,ACI)模型大鼠神经损伤的影响.方法 将大鼠随机分为Sham组、Model组、MogV低剂量组(MogV-L)、MogV高剂量组(MogV-H)、MogV-H+环巴胺(Shh抑制剂)组;对大鼠神经功能进行评分;酶联免疫吸附(Enzyme-linked im-munosorbent assay,ELISA)法检测血清神经生长因子(Nerve growth factor,NGF)、乳酸脱氢酶(Lactic dehy-drogenase,LDH)水平;苏木精-伊红染色法(Hematoxylin-eosin staining,HE)染色观察大鼠海马组织形态;末端脱氧核苷酸转移酶介导的dUTP缺口末端标记技术(Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling,TUNEL)检测大鼠海马神经元凋亡率;氯化三苯基四氮唑(Triphenyltetrazolium chloride,TTC)检测大鼠脑梗死面积;用试剂盒检测脑组织活性氧(Reactive oxygen species,ROS)、丙二醛(Malonalde-hyde,MDA)、超氧化物歧化酶(Superoxide dismutase,SOD)、过氧化氢酶(Catalase,CAT)水平;Western blot检测大鼠海马组织裂解的半胱氨酸蛋白酶-3(Cleaved caspase-3)、髓鞘碱性蛋白(Myelin basic protein,MBP)、少突胶质细胞转录因子(Oligodendrocyte transcription factor 1,Olig1)、Shh、Gli1蛋白表达水平.结果 与Sham组比较,Model组大鼠海马神经组织有明显损伤,神经功能评分、血清LDH水平、细胞凋亡率、脑梗死面积、海马组织中ROS和MDA水平、Cleaved caspase-3蛋白表达水平显著升高,血清NGF水平、海马组织中SOD和CAT活性、Shh,Gli1,MBP,Olig1蛋白表达水平显著降低(P＜0.05);与Model组比较,MogV-L组和MogV-H组大鼠海马组织形态有明显改善,神经功能评分、血清LDH水平、细胞凋亡率、脑梗死面积、海马组织中ROS和MDA水平、Cleaved caspase-3蛋白表达水平显著降低,血清NGF水平、海马组织中SOD和CAT活性、Shh,Gli1,MBP,Olig1蛋白表达水平升高(P＜0.05);环巴胺可减轻MogV对ACI模型大鼠神经损伤的改善作用(P＜0.05).结论 MogV可能通过激活Shh/Gli1信号通路来减轻ACI模型大鼠氧化应激、神经元凋亡和神经损伤,改善神经功能.
Abstract
Objective To investigate the effect of mogroside V(MogV)on nerve injury in rats with a-cute cerebral infarction(ACI)by regulating Sonic hedgehog/glioma-associated oncogene 1(Shh/Gli1)signa-ling pathway.Methods Rats were randomly separated into Sham group,Model group,MogV low-dose group(MogV-L),MogV high-dose group(MogV-H),and MogV-H+cyclophosphamide group(Shh inhibitor).The neural function of rats was scored.Enzyme-linked immunosorbent assay(ELISA)method was applied to detect serum Nerve growth factor(NGF)and Lactic dehydrogenase(LDH)levels.The morphology of rat hip-pocampus was observed by Hematoxylin-eosin staining(HE)staining.Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL was applied to detect the apoptosis rate of hippocampal neurons in rats.Triphenyltetrazolium chloride(TTC)staining method was applied to detect the area of cerebral infarction in rats.Commercial reagent kits were applied to detect levels of Reactive oxygen species(ROS),Malonalde-hyde(MDA),Superoxide dismutase(SOD),and Catalase(CAT)in brain tissue.The protein expression lev-els of Cleaved caspase-3,Myelin basic protein(MBP),Oligodendrocyte transcription factor 1(Olig1),Shh and Glil were detected by Western blot.Results Compared with the Sham group,the model group showed great damage to the hippocampal nerve tissue in rats,the neurological function score,serum LDH level,neu-ronal apoptosis rate,cerebral infarction area,ROS and MDA levels in hippocampal tissue,and the cleaved caspase-3 protein expression level were greatly increased,the serum NGF level,SOD and CAT activities in hippocampal tissue,and the protein expression levels of Shh,Gli1,MBP,and Olig1 were greatly reduced(P＜0.05).Compared with the Model group,the hippocampal morphology of rats in MogV-L and MogV-H groups was significantly improved,the neurological function score,serum LDH level,neuronal apoptosis rate,cerebral infarction area,ROS and MDA levels in hippocampal tissue,and the cleaved caspase-3 protein expres-sion level were greatly reduced,the serum NGF level,SOD and CAT activities in hippocampal tissue,and the Shh,Gli1,MBP,and Olig1 protein expression levels were greatly increased(P＜0.05).Cyclopamine was a-ble to alleviate the improvement effect of MogV on nerve damage in ACI rats(P＜0.05).Conclusion MogV may reduce oxidative stress,neuronal apoptosis and nerve damage in ACI rats to improve nerve function by ac-tivating Shh/Gli1 signaling pathway.
关键词
罗汉果苷V/音猬因子/家族锌指蛋白1信号通路/急性脑梗死/神经损伤Key words
Mogroside V/Shh/Gli1 pathway/Acute cerebral infarction/Neurological injury引用本文复制引用
出版年
2024
NETL
NSTL
万方数据