中华肾脏病杂志
中華腎髒病雜誌
중화신장병잡지
2013年
6期
439-443
,共5页
胡凤琪%袁海%王桂荣%丁国华
鬍鳳琪%袁海%王桂榮%丁國華
호봉기%원해%왕계영%정국화
表面活性蛋白A%表面活性蛋白D%尿路感染%天然免疫
錶麵活性蛋白A%錶麵活性蛋白D%尿路感染%天然免疫
표면활성단백A%표면활성단백D%뇨로감염%천연면역
Surfactant protein A%Surfactant protein D%Urinary tract infection%Innate immunity
目的 研究表面活性蛋白(SP)A及SP-D在尿路感染(UTI)中的作用和机制,评价SP-A及SP-D缺失对UTI的影响.方法 免疫组化评估SP-A及SP-D在野生型(WT)C57BL/6雌性小鼠肾脏内的表达及分布；Western印迹评价SP-A及SP-D双重敲除(SP-A/D KO)与WT小鼠肾脏p38磷酸化水平.应用SP-A/D KO小鼠和WT雌性小鼠构建逆行性小鼠UTI模型,小鼠分为2组:SP-MD KO+ UTI组和WT+ UTI组,分别实验24 h和48 h.进行细菌培养及计数各组肾脏及尿液中的细菌含量,HE染色评价各组病理改变；免疫组化及计数肾脏及尿液内中性粒细胞的数量.体外细菌培养观察SP-A及SP-D对致尿路感染大肠杆菌(UPEC)生长的影响.结果 WT小鼠肾脏内SP-A及SP-D主要分布于近曲小管及集合管.SP-MD KO小鼠肾脏内p38磷酸化水平显著高于WT小鼠(P＜0.05).SP-A/D KO小鼠在发生UTI时,其肾脏病理改变更重,中性粒细胞浸润更多.SP-A/D KO小鼠肾脏内及尿液内细菌量在24 h及48 h均显著高于WT小鼠(均P＜ 0.01).体外实验显示,SP-A及SP-D均可显著抑制UPEC的生长(均P＜ 0.05).结论 肾脏内表达SP-A及SP-D；SP-A/D KO小鼠与WT小鼠相比,其对UTI更易感,其机制可能与UTI时炎性因子分泌不足及SP-A和SP-D对致尿路感染大肠杆菌生长的直接抑制有关.
目的 研究錶麵活性蛋白(SP)A及SP-D在尿路感染(UTI)中的作用和機製,評價SP-A及SP-D缺失對UTI的影響.方法 免疫組化評估SP-A及SP-D在野生型(WT)C57BL/6雌性小鼠腎髒內的錶達及分佈；Western印跡評價SP-A及SP-D雙重敲除(SP-A/D KO)與WT小鼠腎髒p38燐痠化水平.應用SP-A/D KO小鼠和WT雌性小鼠構建逆行性小鼠UTI模型,小鼠分為2組:SP-MD KO+ UTI組和WT+ UTI組,分彆實驗24 h和48 h.進行細菌培養及計數各組腎髒及尿液中的細菌含量,HE染色評價各組病理改變；免疫組化及計數腎髒及尿液內中性粒細胞的數量.體外細菌培養觀察SP-A及SP-D對緻尿路感染大腸桿菌(UPEC)生長的影響.結果 WT小鼠腎髒內SP-A及SP-D主要分佈于近麯小管及集閤管.SP-MD KO小鼠腎髒內p38燐痠化水平顯著高于WT小鼠(P＜0.05).SP-A/D KO小鼠在髮生UTI時,其腎髒病理改變更重,中性粒細胞浸潤更多.SP-A/D KO小鼠腎髒內及尿液內細菌量在24 h及48 h均顯著高于WT小鼠(均P＜ 0.01).體外實驗顯示,SP-A及SP-D均可顯著抑製UPEC的生長(均P＜ 0.05).結論 腎髒內錶達SP-A及SP-D；SP-A/D KO小鼠與WT小鼠相比,其對UTI更易感,其機製可能與UTI時炎性因子分泌不足及SP-A和SP-D對緻尿路感染大腸桿菌生長的直接抑製有關.
목적 연구표면활성단백(SP)A급SP-D재뇨로감염(UTI)중적작용화궤제,평개SP-A급SP-D결실대UTI적영향.방법 면역조화평고SP-A급SP-D재야생형(WT)C57BL/6자성소서신장내적표체급분포；Western인적평개SP-A급SP-D쌍중고제(SP-A/D KO)여WT소서신장p38린산화수평.응용SP-A/D KO소서화WT자성소서구건역행성소서UTI모형,소서분위2조:SP-MD KO+ UTI조화WT+ UTI조,분별실험24 h화48 h.진행세균배양급계수각조신장급뇨액중적세균함량,HE염색평개각조병리개변；면역조화급계수신장급뇨액내중성립세포적수량.체외세균배양관찰SP-A급SP-D대치뇨로감염대장간균(UPEC)생장적영향.결과 WT소서신장내SP-A급SP-D주요분포우근곡소관급집합관.SP-MD KO소서신장내p38린산화수평현저고우WT소서(P＜0.05).SP-A/D KO소서재발생UTI시,기신장병리개변경중,중성립세포침윤경다.SP-A/D KO소서신장내급뇨액내세균량재24 h급48 h균현저고우WT소서(균P＜ 0.01).체외실험현시,SP-A급SP-D균가현저억제UPEC적생장(균P＜ 0.05).결론 신장내표체SP-A급SP-D；SP-A/D KO소서여WT소서상비,기대UTI경역감,기궤제가능여UTI시염성인자분비불족급SP-A화SP-D대치뇨로감염대장간균생장적직접억제유관.
Objective To investigate the role of surfactant protein (SP)-A and SP-D in urinary tract infection mouse model,and evaluate the effects of SP-A and SP-D absence on urinary tract infection.Methods SP-A and SP-D double knockout (SP-A/D KO) mice were made.SP-A/D KO and wild-type (WT) C57BL/6 female mice were used for this study.The expression of SP-A and SP-D in kidney was detected by immunohistochemistry (IHC).The levels of p-p38 and p38 protein in kidneys were measured by Western blotting.Uropathogenic Escherichia coli or buffer was delivered into the bladder of female mice.At 24 and 48 h after inoculation,CFU of Escherichia coli in the kidney and urine of the treated and control mice were measured.Histological,cellular and molecular analysis were performed by several methods of H/E staining,IHC and Western blotting.The effects of SP-A and SP-D on bacterial growth were studied in vitro.Results SP-A and SP-D in kidney were located in the proximal tubules and collecting tubules.Compared with WT mice,infected SP-A/D KO mice with UPEC had higher CFU in kidneys and urine at 24 h and 48 h,increased inflammatory cells infiltration in kidneys (P＜0.05).Compared with WT mice,SP-A/D KO mice had higher p38 MAPK phosphorylation levels in kidneys (P ＜ 0.05).Growth of Escherichia coli was greatly inhibited by both SP-A and SP-D (P＜0.05).Conclusions Both SP-A and SP-D are expressed in kidney.SP-A and SP-D can attenuate UTI induced by UPEC which may be through inhibiting bacterial growth and modulating renal inflammation.
