基于PERK信号通路调控ERS介导神经元凋亡探讨脊髓伤方对大鼠脊髓压迫神经元的保护作用及机制  

Exploring the protective effect and mechanism of Jisuishang Formula on rat spinal cord compressed neurons based on the regulation of ERS-mediated neuronal apoptosis by PERK sig⁃nalling pathway

作　　者：刘春志 钟妮[1] 张鼎 汤立蓉 杨汉立 张玲[1] 钟远鸣[3] LIU Chun-zhi;ZHONG Ni;ZHANG Ding;TANG Li-rong;YANG Han-li;ZHANG Ling;ZHONG Yuan-ming(Guangxi University of Chinese Medicine,Nanning Guangxi 530000 China;West China Second University Hospital,Sichuan University,Chengdu Sichuan 610000 China;First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning Guangxi 530000 China)

机构地区：[1]广西中医药大学,广西南宁530000 [2]四川大学华西第二医院,四川成都610000 [3]广西中医药大学第一附属医院,广西南宁530000

出　　处：《时珍国医国药》2024年第13期2905-2911,共7页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(82260942)。

摘　　要：目的观察脊髓伤方对颈脊髓压迫模型大鼠PERK/eIF2α/CHOP信号通路相关蛋白及mRNA表达的影响,探究其通过调控ERS以抑制神经元凋亡可能的效应机制。方法实验分为假手术组、模型组、阳性对照组、脊髓伤方低、中、高剂量组(n=8)。制备脊髓压迫(CSM)模型大鼠。假手术组和模型组以生理盐水灌胃,阳性对照组以GSK2606414溶液腹腔注射、脊髓伤方各剂量实验组予以不同浓度脊髓伤方灌胃,连续4周。术后2周、4周分别观察颈脊髓压迫大鼠模型BBB评分、改良Rivlin斜板评分;术后4周取颈脊髓组织,HE染色观察模型大鼠的组织病理改变;免疫荧光观察组织CHOP、Caspase-3表达情况;rt-qPCR和Western Blotting观察PERK/eIF2α/CHOP信号通路相关蛋白和mRNA的表达水平。结果与假手术组比较,模型组BBB评分和改良Rivlin斜板评分降低,组织中CHOP、Caspase-3蛋白表达明显升高,PERK/eIF2α/CHOP信号通路相关mRNA和蛋白表达水平升高(P<0.05);与模型组比较,阳性对照组及脊髓伤方各剂量实验组BBB评分和改良Rivlin斜板评分明显升高,组织中CHOP、Caspase-3蛋白表达明显降低,PERK/eIF2α/CHOP信号通路相关mRNA和蛋白表达水平降低(P<0.05);脊髓伤方各剂量实验组中中剂量组效果较好(P<0.05)。结论脊髓伤方可通过干预PERK/eIF2α/CHOP信号通路减轻颈脊髓神经元内质网应激,减少脊髓神经元凋亡,从而保护颈脊髓组织,改善颈脊髓受压破大鼠肢体运动功能。Objective To observe the effects of Jisuishang Formula on the expression of mRNAs and proteins related to the PERK/eIF2α/CHOP signalling pathway in cervical spinal cord compression model rats,and to investigate the possible effect mechanisms through the regulation of ERS to inhibit neuronal apoptosis.Methods Spinal cord compression(CSM)model rats were prepared and divided into sham-operated group,model group,positive control group,and low,medium,and high dose group of Jisuishang Formula(n=8).Sa⁃line gavage was used in the sham-operated group and the model group,intraperitoneal injection of GSK2606414 solution was used in the positive control group,and different concentrations of Jisuishang Formula were administered in the traditional Chinese medicine group,and gavage was administered for 4 weeks consecutively.The BBB score and modified Rivlin slant board score of the rat model of cervical spinal cord compression were observed at 2 and 4 weeks after surgery;the cervical spinal cord tissues were taken at 4 weeks after surger⁃y,and HE staining was used to observe the histopathological changes of the model rats;the expression of CHOP and caspase3 in the tis⁃sues was observed by immunofluorescence;the expression levels of mRNA and protein related to the PERK/eIF2α/CHOP signalling pathway were observed by rt-qPCR and WB.mRNA and protein expression levels.Results Compared with the sham-operated group,the BBB score and modified Rivlin slant board score of the model group were reduced,the expression of CHOP and caspase3 proteins in the tissues was significantly increased,and the expression levels of mRNA and protein related to the PERK/eIF2α/CHOP signalling path⁃way were elevated(P<0.05);compared with the model group,the BBB score and modified Rivlin slant board score of the positive con⁃trol group and the traditional Chinese medicine group were significantly increased.The BBB score and modified Rivlin slant board score of the positive control group and the Chinese medicine group were significantly higher,the

关 键 词：脊髓伤方 脊髓型颈椎病 PERK/eIF2α/CHOP信号通路 神经元凋亡 

分 类 号：R285.5[医药卫生—中药学]