摘要
目的研究不同剂量甘草酸苷对幼年癫痫持续状态(SE)大鼠模型海马组织的保护作用。方法出生18~21 d雄性Sprague-Dawley(SD)大鼠采用腹腔注射氯化锂和毛果芸香碱制作SE大鼠模型,按照随机数字表法分为5组:对照组、SE组、SE+低剂量甘草酸苷组、SE+中剂量甘草酸苷组和SE+高剂量甘草酸苷组,腹腔内注入不同剂量的甘草酸苷(20 mg/kg、40 mg/kg和60 mg/kg)。采用酶联免疫吸附试验(ELISA)检测SE大鼠模型血清中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β和IL-6的水平;定量实时PCR(qRT-PCR)检测SE大鼠模型海马组织中TNF-α、IL-1β和IL-6的mRNA表达水平;蛋白印迹法检测海马组织中Bax、Bcl-2和Caspase-3的表达水平;苏木精-伊红(HE)染色和尼氏染色观察神经元损伤情况;末端脱氧核苷酸转移酶介导的原位缺口末端标记法检测神经元的凋亡情况;透射电子显微镜观察线粒体的损伤情况。采用单因素ANOVA法进行各组之间均数检验,然后采用Tukey法进行两两比较。结果与对照组比较,SE大鼠模型血清中TNF-α[(369.69±58.07)ng/L比(75.46±14.64)ng/L]、IL-1β[(242.27±25.23)ng/L比(45.29±5.90)ng/L]和IL-6[(288.15±24.60)ng/L比(46.59±8.80)ng/L均显著上调,差异均有统计学意义(均P<0.05);与SE组比较,低、中、高剂量甘草酸苷均可有效减少SE发作后血清中TNF-α[(216.67±8.31)ng/L、(158.81±5.03)ng/L、(113.69±12.54)ng/L比(369.69±58.07)ng/L]、IL-1β[(131.21±5.50)ng/L、(86.60±7.79)ng/L、(65.06±4.39)ng/L比(242.27±25.23)ng/L]和IL-6[(150.24±9.48)ng/L、(101.70±5.85)ng/L、(91.60±2.81)ng/L比(288.15±24.60)ng/L]的释放水平(均P<0.05)。SE造成大鼠模型海马组织神经元损伤、丢失、凋亡和线粒体损伤,甘草酸苷可以改善海马组织中神经元损伤、凋亡和线粒体的损伤。与对照组比较,SE组海马组织中Bax(0.57±0.01比0.14±0.01)和Caspase-3(0.54±0.00比0.11±0.01)水平均显著升高,Bcl-2表达水平(0.27±0.01比0.57±0.02)下降,差异均有统计学意义(均P<0.05);与SE组比较,低、中、高剂量甘草酸苷均可下调Bax(0.51±0.02、0.45±0.03、0.40±0.02比0.57±0.01)和Caspase-3(0.47±0.02、0.42±0.02、0.37±0.01比0.54±0.00)的表达水平,上调Bcl-2(0.41±0.02、0.45±0.02、0.51±0.01比0.27±0.01)的表达水平(均P<0.05)。结论甘草酸苷可以有效保护幼年SE大鼠模型的海马组织。
Objective To study the protective effects of various doses of Glycyrrhizin on hippocampus of young rats with status epilepticus(SE).Methods Lithium chloride and pilocarpine were injected intraperitoneally into male Sprague-Dawley(SD)rats(with a postnatal age of 18-21 days),so as to induce SE in rats.The rats were divided into 5 groups according to the random number table method:control group,SE group,SE+low dose Glycyrrhizin group,SE+medium dose Glycyrrhizin group and SE+high dose Glycyrrhizin group.Three different doses of Glycyrrhizin(20 mg/kg,40 mg/kg and 60 mg/kg)were injected intraperitoneally into the rats.The levels of tumor necrosis factor(TNF)-α,interleukin(IL)-1βand IL-6 in serum of SE rats were determined by enzyme linked immunosorbent assay.Quantitative real-time PCR(qRT-PCR)was used to detect the mRNA expression levels of TNF-α,IL-1βand IL-6 in hippocampus of SE rats.The expression levels of Bax,Bcl-2 and Caspase-3 in hippocampus were detected by Western blot.The damage of neurons was measured by hematoxylin and eosin(HE)staining and Nissl staining.Neurons apoptosis was examined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL).The mitochondrial changes were observed under transmission electron microscopy.One-way ANOVA followed by Tukey post-hoc test was used for statistical analysis.Results Compared to the control group,TNF-α[(369.69±58.07)ng/L vs.(75.46±14.64)ng/L],IL-1β[(242.27±25.23)ng/L vs.(45.29±5.90)ng/L]and IL-6[(288.15±24.60)ng/L vs.(46.59±8.80)ng/L]in the serum of SE rats were significantly up-regulated(all P<0.05).Compared to SE group,low,medium and high doses Glycyrrhizin could effectively reduce the levels of TNF-α[(216.67±8.31)ng/L,(158.81±5.03)ng/L and(113.69±12.54)ng/L vs.(369.69±58.07)ng/L],IL-1β[(131.21±5.50)ng/L,(86.60±7.79)ng/L and(65.06±4.39)ng/L vs.(242.27±25.23)ng/L]and IL-6[(150.24±9.48)ng/L,(101.70±5.85)ng/L and(91.60±2.81)ng/L vs.(288.15±24.60)ng/L]released in serum after SE occurred(all P<0.05).The neuronal damage,loss,apoptosis and mitochondrial damage were found in the hippocampus of SE rats.Glycyrrhizin could ameliorate these symptoms.Compared to the control group,Bax levels(0.57±0.01 vs.0.14±0.01)and Caspase-3 levels(0.54±0.00 vs.0.11±0.01)in the hippocampus of SE rats were markedly increased,while Bcl-2 levels(0.27±0.01 vs.0.57±0.02)were decreased(all P<0.05).Compared to the SE group,low,medium and high doses Glycyrrhizin could effectively reduce the levels of Bax(0.51±0.02,0.45±0.03 and 0.40±0.02 vs.0.57±0.01)and Caspase-3(0.47±0.02,0.42±0.02 and 0.37±0.01 vs.0.54±0.00),and increase the levels of Bcl-2(0.41±0.02,0.45±0.02 and 0.51±0.01 vs.0.27±0.01)(all P<0.05).Conclusions Glycyrrhizin can effectively protect the hippocampus of young rats with SE.
作者
齐凤芹
李秀敏
马燕
高一博
张波
Qi Fengqin;Li Xiumin;Ma Yan;Gao Yibo;Zhang Bo(Department of Pediatrics,the Second People′s Hospital of Liaocheng(the Second Hospital of Liaocheng Affiliated to Shandong First Medical University),Linqing 252600,China;Department of Emergency,the Second People′s Hospital of Liaocheng(the Second Hospital of Liaocheng Affiliated to Shandong First Medical University),Linqing 252600,China)
出处
《中华实用儿科临床杂志》
CAS
CSCD
北大核心
2022年第18期1407-1412,共6页
Chinese Journal of Applied Clinical Pediatrics
基金
山东省医药卫生科技发展计划项目(2019WS104)。
