摘要
目的观察淫羊藿苷(ICA)对大鼠海马组织中沉默信息调节因子6(SIRT6)的激活作用及对脓毒症相关性脑病(SAE)的影响,并探讨其作用机制.方法将60只成年雄性SD大鼠按随机数字表法分为假手术组(Sham组)、模型组〔脂多糖(LPS组)〕、LPS+ICA组,每组20只.采用尾静脉注射LPS10mg/kg的方法制备脓毒症大鼠模型,Sham组同途径给予等量生理盐水.LPS+ICA组注射LPS后腹腔注射ICA注射液(50mg·kg-1·d-1),每日1次;LPS组及Sham组同途径给予等量生理盐水.于制模后2、4、6d监测大鼠脑电图变化,计算脑电图各波形比例.6d后处死大鼠取脑组织,采用酶联免疫吸附试验(ELISA)检测海马组织中肿瘤坏死因子-α(TNF-α)含量,采用免疫组化法及蛋白质免疫印迹试验(WesternBlot)检测海马组织中SIRT6的表达.结果与Sham组比较,LPS组制模后2d开始脑电图δ波百分比即明显增加,至6d仍明显高于制模前,说明SAE模型制备成功.而LPS+ICA组制模后各时间点脑电图δ波百分比均较LPS组明显减少,以6d差异最为显著〔(12.92±0.62)%比(17.48±0.39)%,P<0.05〕,说明ICA干预有效.LPS组海马组织中TNF-α水平较Sham组明显升高(ng/L:178.40±7.35比12.76±0.93,P<0.05),SIRT6蛋白水平较Sham组明显下降(SIRT6/β-actin:0.62±0.03比1.35±0.05,P<0.05);LPS+ICA组海马组织中TNF-α水平较LPS组明显下降(ng/L:82.31±7.97比178.40±7.35,P<0.05),SIRT6蛋白水平较LPS组明显升高(SIRT6/β-actin:1.14±0.06比0.62±0.03,P<0.05).结论ICA通过上调海马组织中SIRT6蛋白表达,可减轻海马组织的炎症反应及SAE严重程度.
Objective To observe the effect of icariin(ICA)on the activation of silent information regulator 6(SIRT6)in rat hippocampus and on sepsis associated encephalopathy(SAE),and to explore the possible mechanism.Methods Sixty adult male SD rats were randomly divided into sham operation(Sham)group,model(lipopolysaccharide,LPS)group,and LPS+ICA group,with 20 rats in each group.The rat model of sepsis was made by injecting LPS 10 mg/kg into tail vein.The rats in Sham group were given the same amount of normal saline(NS).After LPS injection,ICA(50 mg·kg-1·d-1)was intraperitoneally injected once daily in LPS+ICA group;the same amount of NS was given in LPS group and Sham group.The electroencephalogram(EEG)changes of rats were monitored on day 2,4 and 6 after model establishment,and the proportion of each waveform was calculated.After 6 days,the rats were killed to take brain tissue.The content of tumor necrosis factor-α(TNF-α)in hippocampus was detected by enzyme linked immunosorbent assay(ELISA),and the expression of SIRT6 in hippocampus was tested by immunehistochemistry and Western Blot.Results Compared with Sham group,the percentage ofδwave of EEG in LPS group increased significantly after 2 days of modeling,and it was still significantly higher than that before modeling after 6 days of modeling,which indicated that SAE model was successfully prepared.The percentage ofδwave of EEG in LPS+ICA group was significantly lower than that in LPS group at each time point after modeling,and the difference was the most significant on day 6[(12.92±0.62)%vs.(17.48±0.39)%,P<0.05],which indicated that ICA intervention was effective.The level of TNF-αin LPS group was significantly higher than that in Sham group(ng/L:178.40±7.35 vs.12.76±0.93,P<0.05),and the level of SIRT6 protein was significantly lower than that in Sham group(SIRT6/β-actin:0.62±0.03 vs.1.35±0.05,P<0.05).The level of TNF-αin hippocampus of LPS+ICA group was significantly lower than that of LPS group(ng/L:82.31±7.97 vs.178.40±7.35,P<0.05),and the level of SIRT6 protein was significantly higher than that in LPS group(SIRT6/β-actin:1.14±0.06 vs.0.62±0.03,P<0.05).Conclusion ICA can reduce the degree of inflammation and SAE by up-regulating SIRT6 protein expression in hippocampus.
作者
李剑
徐鑫
郭慧
李旭蕊
齐惠军
申张顺
李明泉
李建国
李素彦
Li Jian;Xu Xin;Guo Hui;Li Xurui;Qi Huijun;Shen Zhangshun;Li Mingquan;Li Jianguo;Li Suyan(Department of Emergency,Hebei General Hospital,Shijiazhuang 050000,Hebei,China;Department of General Medicine,Hebei General Hospital,Shijiazhuang 050000,Hebei,China)
出处
《中国中西医结合急救杂志》
CAS
CSCD
北大核心
2020年第2期157-160,共4页
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care
基金
河北省卫计委医学科学研究课题计划项目(20170261,20170262)
河北省科技计划项目(17277720D)
河北省中医药管理局中医药类科研计划课题(2017060)。
