中华实验和临床感染病杂志（电子版）
中華實驗和臨床感染病雜誌（電子版）
중화실험화림상감염병잡지（전자판）
CHINESE JOURNAL OF EXPERIMENTAL AND CLINICAL INFECTIOUS DISEASES(ELECTRONIC VERSION)
2014年
6期
751-755
,共5页
张国兴%李晓华%赵辉%孙霓%孙宇%李秀江
張國興%李曉華%趙輝%孫霓%孫宇%李秀江
장국흥%리효화%조휘%손예%손우%리수강
促红细胞生成素%内毒素%肾损伤
促紅細胞生成素%內毒素%腎損傷
촉홍세포생성소%내독소%신손상
Erythropoietin%Lipopolysaccharide%Kidney injury
目的：利用脂多糖（LPS）建立大鼠内毒素血症导致肾脏损伤模型，探讨促红细胞生成素（EPO）对肾脏损伤的保护作用及可能机制，为防治内毒素引起的肾脏损伤提供依据。方法将40只成年Wistar大鼠随机分成空白对照组、EPO对照组、LPS组和LPS+EPO组。LPS组和LPS+EPO组大鼠尾静脉注射LPS（10 mg/kg）建立肾脏损伤模型，对照组给予同等量生理盐水，30 min后，EPO对照组和LPS+EPO组给予rhEPO（5000 U/kg）经尾静脉注射。其余两组大鼠给予生理盐水。在LPS注射后的6 h和24 h每组分别处死5只大鼠，生化分析仪测定大鼠血清尿素氮（BUN）和肌酐（Cr）水平，放射免疫方法测定大鼠血清肿瘤坏死因子-α（TNF-α）水平。注射后24 h处死大鼠制备肾组织切片，HE染色后光镜下观察大鼠肾脏病理结构改变，透射电子显微镜观察大鼠肾脏超微结构改变。应用免疫印记方法检测大鼠肾脏丙氨酸氨基转移酶（AKT）、磷酸化丙氨酸氨基转移酶（p-AKT）以及核因子-κB（NF-κB）表达水平。结果与对照组比较，LPS组和LPS+EPO组大鼠血清BUN、Cr和TNF-α水平升高（P均＜0.05）；LPS组以上3个指标升高程度显著高于LPS+EPO组（P均＜0.05）。与对照组比较，LPS组p-AKT和p-AKT/AKT表达增强（p-AKTP=0.000、p-AKT/AKTP=0.000）、NF-κB表达增强（P=0.012）；与LPS组比较，LPS+EPO组p-AKT和p-AKT/AKT表达下降（p-AKTP=0.002、p-AKT/AKTP=0.005），NF-κB表达下降（P=0.066）。光镜下LPS组肾小管上皮细胞坏死、间质水肿和淋巴细胞浸润；LPS+EPO组亦可见间质水肿和淋巴细胞浸润，但较LPS组减轻。电镜下LPS组肾小管上皮细胞空泡化，线粒体固缩和内皮细胞增生；LPS+EPO组肾小球滤过膜增厚，远曲小管上皮细胞内线粒体轻度肿胀和溶酶体增多，损伤较LPS组减轻。结论 EPO可通过减轻炎症反应、减轻组织损伤有效的保护LPS导致的肾损伤，其机制可能与磷脂酰肌醇3-激酶（PI3K）/AKT/NF-κB信号通路有关。
目的：利用脂多糖（LPS）建立大鼠內毒素血癥導緻腎髒損傷模型，探討促紅細胞生成素（EPO）對腎髒損傷的保護作用及可能機製，為防治內毒素引起的腎髒損傷提供依據。方法將40隻成年Wistar大鼠隨機分成空白對照組、EPO對照組、LPS組和LPS+EPO組。LPS組和LPS+EPO組大鼠尾靜脈註射LPS（10 mg/kg）建立腎髒損傷模型，對照組給予同等量生理鹽水，30 min後，EPO對照組和LPS+EPO組給予rhEPO（5000 U/kg）經尾靜脈註射。其餘兩組大鼠給予生理鹽水。在LPS註射後的6 h和24 h每組分彆處死5隻大鼠，生化分析儀測定大鼠血清尿素氮（BUN）和肌酐（Cr）水平，放射免疫方法測定大鼠血清腫瘤壞死因子-α（TNF-α）水平。註射後24 h處死大鼠製備腎組織切片，HE染色後光鏡下觀察大鼠腎髒病理結構改變，透射電子顯微鏡觀察大鼠腎髒超微結構改變。應用免疫印記方法檢測大鼠腎髒丙氨痠氨基轉移酶（AKT）、燐痠化丙氨痠氨基轉移酶（p-AKT）以及覈因子-κB（NF-κB）錶達水平。結果與對照組比較，LPS組和LPS+EPO組大鼠血清BUN、Cr和TNF-α水平升高（P均＜0.05）；LPS組以上3箇指標升高程度顯著高于LPS+EPO組（P均＜0.05）。與對照組比較，LPS組p-AKT和p-AKT/AKT錶達增彊（p-AKTP=0.000、p-AKT/AKTP=0.000）、NF-κB錶達增彊（P=0.012）；與LPS組比較，LPS+EPO組p-AKT和p-AKT/AKT錶達下降（p-AKTP=0.002、p-AKT/AKTP=0.005），NF-κB錶達下降（P=0.066）。光鏡下LPS組腎小管上皮細胞壞死、間質水腫和淋巴細胞浸潤；LPS+EPO組亦可見間質水腫和淋巴細胞浸潤，但較LPS組減輕。電鏡下LPS組腎小管上皮細胞空泡化，線粒體固縮和內皮細胞增生；LPS+EPO組腎小毬濾過膜增厚，遠麯小管上皮細胞內線粒體輕度腫脹和溶酶體增多，損傷較LPS組減輕。結論 EPO可通過減輕炎癥反應、減輕組織損傷有效的保護LPS導緻的腎損傷，其機製可能與燐脂酰肌醇3-激酶（PI3K）/AKT/NF-κB信號通路有關。
목적：이용지다당（LPS）건립대서내독소혈증도치신장손상모형，탐토촉홍세포생성소（EPO）대신장손상적보호작용급가능궤제，위방치내독소인기적신장손상제공의거。방법장40지성년Wistar대서수궤분성공백대조조、EPO대조조、LPS조화LPS+EPO조。LPS조화LPS+EPO조대서미정맥주사LPS（10 mg/kg）건립신장손상모형，대조조급여동등량생리염수，30 min후，EPO대조조화LPS+EPO조급여rhEPO（5000 U/kg）경미정맥주사。기여량조대서급여생리염수。재LPS주사후적6 h화24 h매조분별처사5지대서，생화분석의측정대서혈청뇨소담（BUN）화기항（Cr）수평，방사면역방법측정대서혈청종류배사인자-α（TNF-α）수평。주사후24 h처사대서제비신조직절편，HE염색후광경하관찰대서신장병리결구개변，투사전자현미경관찰대서신장초미결구개변。응용면역인기방법검측대서신장병안산안기전이매（AKT）、린산화병안산안기전이매（p-AKT）이급핵인자-κB（NF-κB）표체수평。결과여대조조비교，LPS조화LPS+EPO조대서혈청BUN、Cr화TNF-α수평승고（P균＜0.05）；LPS조이상3개지표승고정도현저고우LPS+EPO조（P균＜0.05）。여대조조비교，LPS조p-AKT화p-AKT/AKT표체증강（p-AKTP=0.000、p-AKT/AKTP=0.000）、NF-κB표체증강（P=0.012）；여LPS조비교，LPS+EPO조p-AKT화p-AKT/AKT표체하강（p-AKTP=0.002、p-AKT/AKTP=0.005），NF-κB표체하강（P=0.066）。광경하LPS조신소관상피세포배사、간질수종화림파세포침윤；LPS+EPO조역가견간질수종화림파세포침윤，단교LPS조감경。전경하LPS조신소관상피세포공포화，선립체고축화내피세포증생；LPS+EPO조신소구려과막증후，원곡소관상피세포내선립체경도종창화용매체증다，손상교LPS조감경。결론 EPO가통과감경염증반응、감경조직손상유효적보호LPS도치적신손상，기궤제가능여린지선기순3-격매（PI3K）/AKT/NF-κB신호통로유관。
Objective To investigate the protective effect of erythropoietin (EPO) on kidney injuries and the possible mechanisms in rats with kidney injury induced by lipopolysaccharide (LPS). Methods Total of 40 adult Wistar rats were randomly divided into four experimental groups:blank control group, EPO control group, LPS group and (LPS+EPO) group. Rats model of kidney injury were established by tail vein injection of LPS for 10 mg/kg in LPS group and (LPS+EPO) group, while the control groups received the same amount of saline. Thirty minutes later, recombinant human EPO treatment (5 000 U/kg) was administered by tail vein injection in (LPS+EPO) group and EPO control group, while saline was administered in the other two groups. Six hours and 24 hours after LPS challenge, the changes of blood urea nitrogen (BUN) and creatinine (Cr) were evaluated by biochemical analysis and the levels of tumor necrosis factor-α(TNF-α) were determined by immunoradioassay. Twenty-four hours after the treatment, histological examination of tissue sections was carried out by hematoxylin and eosin staining, while ultrastructure evaluation of organ tissues was assessed by transmission electron microscopy. The expression levels of alanine aminotransferase assav (AKT), phosphorylation of alanine aminotransferase assav (p-AKT) and nuclear factor-кB (NF-κB) were detected by Western blot. Results Compared with the control group, the serum BUN, Cr and TNF-αlevels of rats in LPS group and (LPS+EPO) group were elevated (P<0.05);among which the above three index in LPS group increased signiifcantly than that in (LPS+EPO) group (P<0.05). Compared with the control group, the levels of p-AKT and p-AKT/AKT expression enhanced in LPS group (p-AKTP=0.000, p-AKT/AKTP=0.000), while NF-κB expression increased (P=0.012). Compared with LPS group, the levels of p-AKT and p-AKT/AKT expression decreased in (LPS+EPO) group (p-AKTP=0.002, p-AKT/AKTP=0.005), while NF-κB expression expression decreased (P=0.066). Light microscope showed that tubular epithelial cell necrosis, interstitial edema and lymphocytic inifltration in LPS group. Interstitial edema and lymphocytic inifltration were also found in (LPS + EPO) group, but less seriously than that in LPS group. Electron microscope showed that tubular epithelial cells vacuoles, mitochondria condensation and endothelial cell proliferation in LPS group. Glomerular ifltration membrane thickening, increased distal convoluted tubule epithelium mild swelling of mitochondria and lysosomes were also found in (LPS+EPO) group, but less seriously compared with that in LPS group. Conclusions EPO may play a protective role against LPS-induced kidney injuries by reducing the inflammatory response and tissue degeneration, possibly via the phosphatidylinositol 3-kinase/AKT and NF-κB signaling pathways.
