持续去甲肾上腺素输注对早期脓毒症大鼠肾脏的保护作用及其机制研究
지속거갑신상선소수주대조기농독증대서신장적보호작용급기궤제연구
Renal protective effect and mechanism of continuous infusion of norepinephrine in rat with early stage sepsis
  • 万方数据
    中国小儿急救医学 중국소인급구의학
    CHINESE PEDIATRIC EMERGENCY MEDICINE
    2014年 7期 401-407 ,共7页

    梁漂红%曾其毅

    량표홍%증기의

    脓毒症%急性肾损害%去甲肾上腺素%细胞因子%线粒体%氧化应激 膿毒癥%急性腎損害%去甲腎上腺素%細胞因子%線粒體%氧化應激
    농독증%급성신손해%거갑신상선소%세포인자%선립체%양화응격
    Sepsis%Acute kidney injury%Norepinephrine%Cytokines%Mitochondria%Oxidative stress
    目的 探讨持续去甲肾上腺素输注对早期脓毒症大鼠肾脏是否存在保护作用及其可能机制.方法 30只SPF级健康雄性SD大鼠采用随机数字表法随机分为5组(6只/组).对照组:腹腔注射生理盐水,并开始持续输注生理盐水1 ml/h.脂多糖(lipopolysaccharide,LPS)组和低剂量、中剂量和高剂量去甲肾上腺素组:腹腔注射LPS 10 mg/kg,LPS组持续输注生理盐水1 ml/h,低、中、高剂量组分别持续输注0.06、0.30、0.60 μg/(kg· min)去甲肾上腺素溶液1 ml/h,均持续输注24 h处死大鼠.检测2、6h大鼠血清炎症因子肿瘤坏死因子(tumor necrosis factor,TNF)-α、白细胞介素(interleukin,IL)-1β、IL-6及IL-10水平,24h大鼠血清C-反应蛋白(CRP)、肌酐(Cr)和尿素氮(BUN)水平,膜电位和氧化应激相关指标,电镜观察肾脏线粒体肿胀程度,显微镜观察肾脏病理变化.结果 与对照组比较,2 h LPS组TNF-α浓度为(2 203.3±1 028.7) pg/ml、IL-1β (2214.5±457.0) pg/ml、IL-6(7 784.7±248.2)pg/ml及IL-10(1 076.1±368.4) pg/ml,均明显升高(P＜ 0.05);24 h CRP(0.35±0.24) mg/L,Cr(30.8±11.5)μmol/L,BUN(7.7±1.8) mmol/L,一氧化氮(1 057.4±172.9) μmol/gprot,均明显升高(P＜0.01),肾脏线粒体膜电位0.0464±0.0185,明显下降(P＜0.01).与LPS组比较,2h低剂量组TNF-α浓度为(506.8±301.7) pg/ml、IL-1β(415.6±178.0) pg/ml及IL-10(381.7±171.0) pg/ml,均明显下降(P＜0.05),24 h BUN下降为(5.2±1.4) mmol/L(P ＜0.05),线粒体膜电位上升为0.3474 ±0.152 6(P ＜0.05);2 h中剂量组TNF-α浓度为(323.9±227.9) pg/ml、IL-1β(700.0 ±246.2)pg/ml,IL-10(282.6±134.4) pg/ml,均明显下降(P＜0.05),24 h CRP下降为(0.17 ±0.08) mg/L(P ＜0.05),线粒体膜电位上升为0.3775±0.143 7(P ＜0.05);高剂量组2hTNF-α浓度为(378.7±89.8)pg/ml、IL-1β(945.7±264.4) pg/ml,明显下降(P＜0.05),24 h CRP(0.19 ±0.12) mg/L、Cr(23.2±3.4)μmol/L均明显下降(P＜0.05).电镜观察显示LPS组线粒体包膜模糊及基质空泡化、凝固,显微镜观察显示LPS组肾组织间隙水肿,单核-巨噬细胞浸润,肾小球皱缩,肾小管上皮细胞肿胀、空泡变性等,中剂量组病理损伤明显减轻.结论 持续去甲肾上腺素输注对早期脓毒症大鼠肾脏有一定的保护作用,其机制与降低炎症因子风暴的水平、减轻氧化应激损害、改善线粒体功能相关.
    목적 탐토지속거갑신상선소수주대조기농독증대서신장시부존재보호작용급기가능궤제.방법 30지SPF급건강웅성SD대서채용수궤수자표법수궤분위5조(6지/조).대조조:복강주사생리염수,병개시지속수주생리염수1 ml/h.지다당(lipopolysaccharide,LPS)조화저제량、중제량화고제량거갑신상선소조:복강주사LPS 10 mg/kg,LPS조지속수주생리염수1 ml/h,저、중、고제량조분별지속수주0.06、0.30、0.60 μg/(kg· min)거갑신상선소용액1 ml/h,균지속수주24 h처사대서.검측2、6h대서혈청염증인자종류배사인자(tumor necrosis factor,TNF)-α、백세포개소(interleukin,IL)-1β、IL-6급IL-10수평,24h대서혈청C-반응단백(CRP)、기항(Cr)화뇨소담(BUN)수평,막전위화양화응격상관지표,전경관찰신장선립체종창정도,현미경관찰신장병리변화.결과 여대조조비교,2 h LPS조TNF-α농도위(2 203.3±1 028.7) pg/ml、IL-1β (2214.5±457.0) pg/ml、IL-6(7 784.7±248.2)pg/ml급IL-10(1 076.1±368.4) pg/ml,균명현승고(P＜ 0.05);24 h CRP(0.35±0.24) mg/L,Cr(30.8±11.5)μmol/L,BUN(7.7±1.8) mmol/L,일양화담(1 057.4±172.9) μmol/gprot,균명현승고(P＜0.01),신장선립체막전위0.0464±0.0185,명현하강(P＜0.01).여LPS조비교,2h저제량조TNF-α농도위(506.8±301.7) pg/ml、IL-1β(415.6±178.0) pg/ml급IL-10(381.7±171.0) pg/ml,균명현하강(P＜0.05),24 h BUN하강위(5.2±1.4) mmol/L(P ＜0.05),선립체막전위상승위0.3474 ±0.152 6(P ＜0.05);2 h중제량조TNF-α농도위(323.9±227.9) pg/ml、IL-1β(700.0 ±246.2)pg/ml,IL-10(282.6±134.4) pg/ml,균명현하강(P＜0.05),24 h CRP하강위(0.17 ±0.08) mg/L(P ＜0.05),선립체막전위상승위0.3775±0.143 7(P ＜0.05);고제량조2hTNF-α농도위(378.7±89.8)pg/ml、IL-1β(945.7±264.4) pg/ml,명현하강(P＜0.05),24 h CRP(0.19 ±0.12) mg/L、Cr(23.2±3.4)μmol/L균명현하강(P＜0.05).전경관찰현시LPS조선립체포막모호급기질공포화、응고,현미경관찰현시LPS조신조직간극수종,단핵-거서세포침윤,신소구추축,신소관상피세포종창、공포변성등,중제량조병리손상명현감경.결론 지속거갑신상선소수주대조기농독증대서신장유일정적보호작용,기궤제여강저염증인자풍폭적수평、감경양화응격손해、개선선립체공능상관.
    Objective To investigate the protective effect and possible mechanisms of continuous infusion of norepinephrine in kidney of septic rats in the early stage.Methods Thirty healthy male SD rats of SPF level were randomly divided into five groups.Rats in control group were given intraperitoneal injection of saline and began a continuous infusion of saline (1 ml/h).Rats in LPS group and the intervention group (low-dose,medium-dose and high-dose norepinephrine group) were given intraperitoneal injection of LPS 10 mg/kg.LPS group began a continuous infusion of saline (1 ml/h) while low,medium and high dose groups began continuous infusion of different norepinephrine solution [(0.06,0.3,0.6 μg/(kg·min)].Rats were sacrificed after 24 hours infusion.We detected serum inflammatory cytokines [tumor necrosis factor (TNF)-α,interleukin(IL)-1β,IL-6 and IL-10] in 2 h and 6 h by ELISA.Rat serum CRP,Cr and BUN,swelling and membrane potential of kidney mitochondria and oxidative stress-related indicators were tested in 24 h.We also observed renal pathologic changes by electronic microscopy and biopsy.Results Compared with the control group,serum levels of TNF-α [(2 203.3 ± 1 028.7) pg/ml],IL-1β [(2 214.5 ±457.0) pg/ml],IL-6 [(7784.7 ±248.2) pg/ml] and IL-10 [(1 076.1 ±368.4) pg/ml] were statistically higher in LPS group in 2 h (P ＜ 0.05) ; CRP [(0.35 ± 0.24) mg/L],Cr [(30.8 ± 11.5) μ mol/L],BUN [(7.7 ± 1.8) mmol/L],NO [(1 057.4 ± 172.9) μmol/gprot] were statistically higher (P ＜ 0.01),membrane potential of kidney mitochondria (0.0464 ±0.018 5) decreased statistically (P ＜0.01).Compared with LPS group,serum levels of TNF-αt [(506.8 ±301.7) pg/ml],IL-lβ [(415.6 ± 178.0) pg/ml],and IL-10 [(381.7 ± 171.0) pg/ml] significantly decreased in low-dose group in 2 h (P ＜0.05),BUN [(5.2 ± 1.4)mmol/L] decreased (P ＜ 0.05),mitochondrial membrane potential (0.347 4 ± 0.152 6) increased in 24 h (P ＜ 0.05) ; serum levels of TNF-α [(323.9 ± 227.9) pg/ml],IL-1 β [(700.0 ± 246.2) pg/ml],and IL-10[(282.6 ± 134.4) pg/ml] significantly decreased statistically in medium-dose group in 2 h (P ＜0.05),CRP [(0.17 ± 0.08) mg/L] decreased statistically (P ＜ 0.01),mitochondrial membrane potential (0.377 5 ±0.143 7) increased in 24 h (P ＜0.05) ;serum levels of TNF-α [(378.7 ±89.8) pg/ml],IL-1β [(945.7 ±264.4) pg/ml] significantly decreased in high-dose group in 2 h (P ＜0.05),CRP [(0.19 ±0.12) mg/L] and Cr [(23.2 ±3.4) μmol/L] decreased in 24 h (P ＜0.05).Mitochondrial matrix coated fuzzy,vacuoles and coagulation were found in LPS group by electronic microscopy examination.Interstitial edema,monocyte-macrophage infiltration,glomerular shrinkage,tubular epithelial cell swelling,empty bubble degeneration were found in LPS group by microscopy examination.Pathological damage was alleviated in mediumdose group.Conclusion Continuous infusion of norepinephrine plays a protective role on renal function in rats with sepsis in the early stage.The intervention protect rat kidney by reducing levels of inflammatory cytokines,oxidative stress and mitochondrial damage.
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