赣南医学院学报
贛南醫學院學報
공남의학원학보
JOURNAL OF GANNAN MEDICAL COLLEGE
2014年
3期
335-340
,共6页
李嵩山%吴明%曾晶晶%李明利%吴乐锋%冯永文
李嵩山%吳明%曾晶晶%李明利%吳樂鋒%馮永文
리숭산%오명%증정정%리명리%오악봉%풍영문
肢体缺血后处理%失血性休克%急性肾损伤%核因子 κB%炎症反应%诱导性一氧化氮合酶
肢體缺血後處理%失血性休剋%急性腎損傷%覈因子 κB%炎癥反應%誘導性一氧化氮閤酶
지체결혈후처리%실혈성휴극%급성신손상%핵인자 κB%염증반응%유도성일양화담합매
Limb remote ischemic postconditioning%hemorrhagic shock%acute kidney injury%inflammatory mediators%ni-tric oxide synthase
目的：探讨肢体缺血后处理对孕兔失血性休克肾损伤的保护及可能的作用机制。方法：24只孕兔制成重度失血性休克模型，均游离双侧股动脉，随机分为假手术组（S 组）、缺血再灌注组（IR 组）、肢体缺血后处理组（LRIP 组）（每组8只）。S 组：游离双侧股动脉后无失血及输血；IR：孕兔股动脉放血使平均动脉压40 mmHg 维持180 min 后，1 h 内匀速回输全部血液，使孕兔血压维持在 MAP 稳定在80 mmHg 之上并平稳24 h；LRIP 组：在急性失血性休克180 min 后，给予夹闭双侧股动脉缺血30 s 再灌注30 s 共10次后（即缺血后处理），再输血复苏至24 h。分别采用全自动生化分析仪测定血尿素氮（BUN）、血肌酐（Cr）水平，酶联免疫吸附试验（ELISA）法测定不同时点（0 h、8 h、16 h、24 h）血清 TNF-α、IL-10的含量，免疫组化法检测肾组织中核因子κB（NF-κB）表达；RT-PCR 法检测肾组织内 TNF-α、IL-10及 iNOS mRNA 的表达；HE 染色观察肾脏病理学改变。结果：（1）与 I/ R 组相比，LRIP 组不同时点 Cr、BUN 浓度降低，血清 TNF-α含量降低、IL-10含量升高，肾组织 NF-κB 表达减少（均<0.05）；（2）与 S 组比较，肾内 TNF-α、IL-10、iNOS mRNA 在 I/ R 组、LRIP 组表达均上调（<0.05或<0.01），与I/ R 组相比，肾内 TNF-α、iNOS mRNA 表达在 LRIP 组下调，IL-10 mRNA 表达上调（<0.05）；（3）S 组肾脏未发生明显的病理改变，I/ R 组肾小管可见明显玻璃样改变和坏死，肾小管略扩张，肾小球和肾小管周围可见炎症细胞浸润，LRIP 组肾脏组织结构正常，少量的炎症细胞浸润，肾小管上皮细胞相对完整，水肿减轻。结论：重复10次缺血30 s再灌注30 s 的肢体后处理对肾脏有保护作用，其机制可能与抑制肾组织 NF-κB 转录、减少 iNOS mRNA的表达、改善肾内炎症反应有关。
目的：探討肢體缺血後處理對孕兔失血性休剋腎損傷的保護及可能的作用機製。方法：24隻孕兔製成重度失血性休剋模型，均遊離雙側股動脈，隨機分為假手術組（S 組）、缺血再灌註組（IR 組）、肢體缺血後處理組（LRIP 組）（每組8隻）。S 組：遊離雙側股動脈後無失血及輸血；IR：孕兔股動脈放血使平均動脈壓40 mmHg 維持180 min 後，1 h 內勻速迴輸全部血液，使孕兔血壓維持在 MAP 穩定在80 mmHg 之上併平穩24 h；LRIP 組：在急性失血性休剋180 min 後，給予夾閉雙側股動脈缺血30 s 再灌註30 s 共10次後（即缺血後處理），再輸血複囌至24 h。分彆採用全自動生化分析儀測定血尿素氮（BUN）、血肌酐（Cr）水平，酶聯免疫吸附試驗（ELISA）法測定不同時點（0 h、8 h、16 h、24 h）血清 TNF-α、IL-10的含量，免疫組化法檢測腎組織中覈因子κB（NF-κB）錶達；RT-PCR 法檢測腎組織內 TNF-α、IL-10及 iNOS mRNA 的錶達；HE 染色觀察腎髒病理學改變。結果：（1）與 I/ R 組相比，LRIP 組不同時點 Cr、BUN 濃度降低，血清 TNF-α含量降低、IL-10含量升高，腎組織 NF-κB 錶達減少（均<0.05）；（2）與 S 組比較，腎內 TNF-α、IL-10、iNOS mRNA 在 I/ R 組、LRIP 組錶達均上調（<0.05或<0.01），與I/ R 組相比，腎內 TNF-α、iNOS mRNA 錶達在 LRIP 組下調，IL-10 mRNA 錶達上調（<0.05）；（3）S 組腎髒未髮生明顯的病理改變，I/ R 組腎小管可見明顯玻璃樣改變和壞死，腎小管略擴張，腎小毬和腎小管週圍可見炎癥細胞浸潤，LRIP 組腎髒組織結構正常，少量的炎癥細胞浸潤，腎小管上皮細胞相對完整，水腫減輕。結論：重複10次缺血30 s再灌註30 s 的肢體後處理對腎髒有保護作用，其機製可能與抑製腎組織 NF-κB 轉錄、減少 iNOS mRNA的錶達、改善腎內炎癥反應有關。
목적：탐토지체결혈후처리대잉토실혈성휴극신손상적보호급가능적작용궤제。방법：24지잉토제성중도실혈성휴극모형，균유리쌍측고동맥，수궤분위가수술조（S 조）、결혈재관주조（IR 조）、지체결혈후처리조（LRIP 조）（매조8지）。S 조：유리쌍측고동맥후무실혈급수혈；IR：잉토고동맥방혈사평균동맥압40 mmHg 유지180 min 후，1 h 내균속회수전부혈액，사잉토혈압유지재 MAP 은정재80 mmHg 지상병평은24 h；LRIP 조：재급성실혈성휴극180 min 후，급여협폐쌍측고동맥결혈30 s 재관주30 s 공10차후（즉결혈후처리），재수혈복소지24 h。분별채용전자동생화분석의측정혈뇨소담（BUN）、혈기항（Cr）수평，매련면역흡부시험（ELISA）법측정불동시점（0 h、8 h、16 h、24 h）혈청 TNF-α、IL-10적함량，면역조화법검측신조직중핵인자κB（NF-κB）표체；RT-PCR 법검측신조직내 TNF-α、IL-10급 iNOS mRNA 적표체；HE 염색관찰신장병이학개변。결과：（1）여 I/ R 조상비，LRIP 조불동시점 Cr、BUN 농도강저，혈청 TNF-α함량강저、IL-10함량승고，신조직 NF-κB 표체감소（균<0.05）；（2）여 S 조비교，신내 TNF-α、IL-10、iNOS mRNA 재 I/ R 조、LRIP 조표체균상조（<0.05혹<0.01），여I/ R 조상비，신내 TNF-α、iNOS mRNA 표체재 LRIP 조하조，IL-10 mRNA 표체상조（<0.05）；（3）S 조신장미발생명현적병리개변，I/ R 조신소관가견명현파리양개변화배사，신소관략확장，신소구화신소관주위가견염증세포침윤，LRIP 조신장조직결구정상，소량적염증세포침윤，신소관상피세포상대완정，수종감경。결론：중복10차결혈30 s재관주30 s 적지체후처리대신장유보호작용，기궤제가능여억제신조직 NF-κB 전록、감소 iNOS mRNA적표체、개선신내염증반응유관。
s:Objectiye:To investigate the protective effects of limb ischemic postconditioning on kidney injury induced by hemorrhagic shock in pregnant rabbits. Methods:The model of severe hemorrhagk shock was established in 24 pregnant rabbits. The rabbits were divided into three groups:Sham operation group(S group),ischemia reperfusion group(IR) and limb remote ischaemic postconditioning group(LRIP)randomly(n = 8). S group:only free bilateral femoral artery,no bleeding and transfusion;IR group:they were bleeding by femoral artery,and maintenance mean arterial pressure 40 mm-Hg for 180 min,then infused all the blood back within 1 hour and maintained MAP over 80 mmHg and stationary stability in 24 hours;LRIP group:they were given the occlusion of bilateral femoral artery,ischemia 30 seconds and reperfusion 30 seconds for 10 times after acute hemorrhagic shock lasting 180 min,then infused blood to recovery lasting 24 h. automatic biochemical analyzer were used respectively to determine blood urea nitrogen(BUN),serum creatinine(Cr),enzyme-linked immunosorbent assay(ELISA)method at different time(0 h,8 h,16 h,24 h)to test the content of serum TNF al-pha,IL-10,immunohistochemical method to detect nuclear factor kappa B in the kidney tissues(NF-kB)expression;RT-PCR method to detect renal tissue TNF alpha,IL-10 and iNOS mRNA expression;HE dyeing observation renal pathologi-cal changes. Results:(1)compared with I/ R group,serum urea nitrogen,creatinine and TNF-α concentration were de-creased,and IL-10 levels was increased in LRIP group at different point,NF-kB expression in kidney tissues decreased(< 0. 05);(2)compared with S group,TNF-α mRNA,IL-10 mRNA,iNOS mRNA expression in kidney tissue were in-creased at I/ R group and LRIP group( < 0. 05 or < 0. 01);compared with I/ R group,TNF-α mRNA and iNOS mR-NA expression decreased,IL-10 mRNA expression increased in renal tissue at LRIP group( < 0. 05);(3)there were no obvious pathological changes in kidney at shame group,and there were obviously hyaline changes,renal tubular necrosis, renal tubules slightly expansion,the inflammatory cell infiltration around glomerular and renal tubular in the I/ R group, there were normal structure in kidney tissue,a small amount of inflammatory cell infiltration,renal tubular epithelial cells relatively intact,edema decreased at LRIP group. Conclusion:ischemia 30 s and reperfusion 30 s repeated 10 times limb remote ischaemic postconditioning has a protective effect on kidney function. Its mechanism may be related to inhibit re-nal tissue NF-κB transcription,decreasing iNOS mRNA expression,improving renal inflammatory response.
