摘要
目的基于S-亚硝基谷胱甘肽还原酶(GSNOR)/S-亚硝基谷胱甘肽(GSNO)途径探讨血必净注射液对大鼠心脏骤停(CA)/心肺复苏(CPR)后神经功能及生存情况影响的潜在机制。方法以室颤法建立CA/CPR大鼠模型,以假手术组大鼠为参照,采用高通量测序分析挖掘两者差异表达基因,并结合酶联免疫吸附测定(ELISA)法检测其海马组织中GSNOR、GSNO含量;筛选血必净注射液活性成分,并与GSNOR进行分子对接。将同法造模成功的大鼠分为模型组(30只)、抑制剂(GSNOR抑制剂)组(30只)、血必净组(30只)、血必净+抑制剂组(30只),并设置假手术组(30只),分别于药物首次干预后3h、24h、3d时进行神经功能评价及生存情况记录,并检测上述时间点各组大鼠海马组织中GSNOR、GSNO含量,分析GSNOR、GSNO含量与大鼠改良的神经系统损害严重程度评分表(mNSS)评分的相关性。结果GSNOR编码基因是模型组与假手术组间的差异表达基因之一;与假手术组比较,模型组大鼠海马组织中GSNOR含量显著升高,而GSNO含量显著降低(P<0.05)。血必净注射液中去甲丹参酮、鼠尾草酚酮等活性成分与GSNOR蛋白的结合能均低于-6 kcal/mol,以氢键连接为主。动物实验结果显示,模型组大鼠各时间点的mNSS评分和海马组织中GSNOR含量均显著高于假手术组(P<0.05),生存率和GSNO含量均显著低于假手术组(P<0.05);各给药组大鼠上述指标均显著改善,且血必净组mNSS评分显著低于抑制剂组,抑制剂组GSNOR、GSNO含量的变化较血必净组更明显,血必净+抑制剂组各指标均显著优于血必净组和抑制剂组(P<0.05)。GSNOR含量与mNSS评分呈正相关,GSNO含量与mNSS评分呈负相关(P<0.05)。结论血必净注射液可以改善CA/CPR后大鼠的神经功能,提高其生存率,该作用可能与下调GSNOR并上调GSNO有关。
OBJECTIVE To explore the potential mechanism of the effect of Xuebijing injection(XBJ)on neurological function and survival of rats after cardiac arrest(CA)/cardiopulmonary resuscitation(CPR)based on the S-nitrosoglutathione reductase(GSNOR)/S-nitrosoglutathione(GSNO)pathway.METHODS The CA/CPR rat model was established by ventricular fibrillation.Using a sham operation group as control,high-throughput sequencing was employed to analyze and mine the differentially expressed genes(DEGs).Enzyme-linked immunosorbent assay was used to determine the contents of GSNOR and GSNO in the hippocampus;the active components of XBJ were screened and subjected to molecular docking analysis with GSNOR.The rats successfully modeled using the same method were divided into model group(n=30),inhibitor(GSNOR inhibitor)group(n=30),XBJ group(n=30)and XBJ+inhibitor group(n=30),and a sham operation group(n=30)was set up.Neurological function was evaluated and survival status was recorded at 3 hours,24 hours and 3 days after the first drug intervention.The contents of GSNOR and GSNO in the hippocampus of rats were determined in each group at the above time points,and the relationship of the contents of GSNOR and GSNO with modified neurologic severity scale(mNSS)score was analyzed.RESULTS GSNOR coding gene was differentially expressed between the model group and the sham operation group.Compared with the sham operation group,GSNOR content increased significantly in the hippocampus of rats in model group,while GSNO content decreased significantly(P<0.05).The active components of XBJ,such as 4-methylenemiltirone and salviolone,could be bound to GSNOR protein,with the binding energy lower than-6 kcal/mol,mainly connected by hydrogen bonds.Animal experiments revealed that mNSS score and GSNOR levels in the hippocampus of rats in the model group were significantly higher than those in the sham operation group(P<0.05),while GSNO levels and survival rate were significantly lower than those in the sham operation group(P<0.05).The above indexes of rats were improved significantly in administration groups,the mNSS score in the XBJ group was significantly lower than that in the inhibitor group,the content changes of GSNOR and GSNO in the inhibitor group were more obvious than those in the XBJ group,and the various indicators in the XBJ+inhibitor group were significantly better than the XBJ group and the inhibitor group(P<0.05).GSNOR content was positively correlated with the mNSS score,and GSNO content was negatively correlated with the mNSS score(P<0.05).CONCLUSIONS XBJ can improve the neurological function of rats and enhance their survival rates after CA/CPR,the mechanism of which may be associated with the down-regulation of GSNOR and the up-regulation of GSNO.
作者
黄德庆
高玉广
张元侃
王政林
邓海霞
黄夏冰
庞延
吴林
HUANG Deqing;GAO Yuguang;ZHANG Yuankan;WANG Zhenglin;DENG Haixia;HUANG Xiabing;PANG Yan;WU Lin(Dept.of Emergency,the First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,China;Scientific Laboratory Centre,Guangxi University of Chinese Medicine,Nanning 530200,China)
出处
《中国药房》
CAS
北大核心
2024年第6期653-658,共6页
China Pharmacy
基金
国家自然科学基金地区科学基金项目(No.818607-89)
广西高校中青年教师科研基础能力提升项目(No.2021KY0281)
广西壮族自治区中医药管理局自筹经费科研课题(No.GXZYZ2021-0191)。
