中华麻醉学杂志
中華痳醉學雜誌
중화마취학잡지
CHINESE JOURNAL OF ANESTHESIOLOGY
2015年
1期
114-118
,共5页
姚翔燕%孟凡民%张加强%杜献慧
姚翔燕%孟凡民%張加彊%杜獻慧
요상연%맹범민%장가강%두헌혜
右美托咪啶%呼吸,人工%内质网%JNK丝裂原活化蛋白激酶类%细胞凋亡
右美託咪啶%呼吸,人工%內質網%JNK絲裂原活化蛋白激酶類%細胞凋亡
우미탁미정%호흡,인공%내질망%JNK사렬원활화단백격매류%세포조망
Dexmedetomidine%Respiration,artifical%Endoplasmic reticulum%JNK mitogen-activated protein kinases%Apoptosis
目的 评价右美托咪定预先给药对大鼠单肺通气(OLV)时内质网应激(ERS)诱导的细胞凋亡及c-Jun氨基末端激酶(JNK)通路的影响.方法 雄性SD大鼠60只,采用随机数字表法分为6组(n=10):假手术组(Sham组)、OLV组、OLV+阿替美唑(α2受体拮抗药)处理组(AD组)、OLV+阿替美唑+右美托咪定处理组(DEX+ AD组)、OLV+右美托咪定低剂量组(DEX-L组)和OLV+右美托咪定高剂量组(DEX-H组).Sham组大鼠行双肺通气2.5h.OLV组行右肺OLV 2.0h、双肺通气0.5h.DEX-L组和DEX-H组分别于OLV前1h静脉输注右美托咪定2.5 μg* kg-1·h-1、5.0μg·kg-1·h-1,1 h完成.AD组于OLV前1h静脉注射阿替美唑250 μg/kg.DEX+AD组于静脉输注DEX 5.0 μg·kg-1 h-1即刻静脉注射阿替美唑250 μg/kg.于双肺通气0.5h时处死大鼠取左肺组织,测定肺组织湿/干重比(W/D),光镜下观察肺组织病理改变并测定肺泡损伤率(IAR),电镜下观察肺组织超微结构,TUNEL法检测肺组织细胞凋亡,RT-PCR和Western blot法分别测定葡萄糖调节蛋白78 (GRP78) mRNA及蛋白、JNK mRNA、磷酸化JNK (p-JNK)蛋白表达水平.结果 与Sham组比较,OLV组、AD组和DEX+AD组W/D、AI和IAR升高(P＜0.01);与OLV组、AD组和DEX+AD组比较,DEX-L组和DEX-H组W/D、AI和IAR降低(P＜0.01).OLV组、AD组和DEX+AD组肺组织结构均发生明显损伤,而DEX-L组和DEX-H组肺组织结构损伤则明显减轻.OLV组、AD组和DEX+AD组有大量肺血管内皮细胞和肺泡上皮细胞发生凋亡,而经DEX干预后细胞凋亡则明显减少.与Sham组比较,OLV组、AD组和DEX+AD组GRP78 mRNA和蛋白、JNK mRNA和p-JNK蛋白表达水平升高(P＜0.01);与OLV组、AD组和DEX+AD组比较,DEX-L组和DEX-H组GRP78 mRNA和蛋白、JNK mRNA和p-JNK蛋白表达水平降低(P＜0.01).结论 右美托咪定预先给药对单肺通气时的肺脏具有保护作用,该作用的可能机制与其抑制JNK信号通路,减轻ERS诱导的细胞凋亡有关.
目的 評價右美託咪定預先給藥對大鼠單肺通氣(OLV)時內質網應激(ERS)誘導的細胞凋亡及c-Jun氨基末耑激酶(JNK)通路的影響.方法 雄性SD大鼠60隻,採用隨機數字錶法分為6組(n=10):假手術組(Sham組)、OLV組、OLV+阿替美唑(α2受體拮抗藥)處理組(AD組)、OLV+阿替美唑+右美託咪定處理組(DEX+ AD組)、OLV+右美託咪定低劑量組(DEX-L組)和OLV+右美託咪定高劑量組(DEX-H組).Sham組大鼠行雙肺通氣2.5h.OLV組行右肺OLV 2.0h、雙肺通氣0.5h.DEX-L組和DEX-H組分彆于OLV前1h靜脈輸註右美託咪定2.5 μg* kg-1·h-1、5.0μg·kg-1·h-1,1 h完成.AD組于OLV前1h靜脈註射阿替美唑250 μg/kg.DEX+AD組于靜脈輸註DEX 5.0 μg·kg-1 h-1即刻靜脈註射阿替美唑250 μg/kg.于雙肺通氣0.5h時處死大鼠取左肺組織,測定肺組織濕/榦重比(W/D),光鏡下觀察肺組織病理改變併測定肺泡損傷率(IAR),電鏡下觀察肺組織超微結構,TUNEL法檢測肺組織細胞凋亡,RT-PCR和Western blot法分彆測定葡萄糖調節蛋白78 (GRP78) mRNA及蛋白、JNK mRNA、燐痠化JNK (p-JNK)蛋白錶達水平.結果 與Sham組比較,OLV組、AD組和DEX+AD組W/D、AI和IAR升高(P＜0.01);與OLV組、AD組和DEX+AD組比較,DEX-L組和DEX-H組W/D、AI和IAR降低(P＜0.01).OLV組、AD組和DEX+AD組肺組織結構均髮生明顯損傷,而DEX-L組和DEX-H組肺組織結構損傷則明顯減輕.OLV組、AD組和DEX+AD組有大量肺血管內皮細胞和肺泡上皮細胞髮生凋亡,而經DEX榦預後細胞凋亡則明顯減少.與Sham組比較,OLV組、AD組和DEX+AD組GRP78 mRNA和蛋白、JNK mRNA和p-JNK蛋白錶達水平升高(P＜0.01);與OLV組、AD組和DEX+AD組比較,DEX-L組和DEX-H組GRP78 mRNA和蛋白、JNK mRNA和p-JNK蛋白錶達水平降低(P＜0.01).結論 右美託咪定預先給藥對單肺通氣時的肺髒具有保護作用,該作用的可能機製與其抑製JNK信號通路,減輕ERS誘導的細胞凋亡有關.
목적 평개우미탁미정예선급약대대서단폐통기(OLV)시내질망응격(ERS)유도적세포조망급c-Jun안기말단격매(JNK)통로적영향.방법 웅성SD대서60지,채용수궤수자표법분위6조(n=10):가수술조(Sham조)、OLV조、OLV+아체미서(α2수체길항약)처리조(AD조)、OLV+아체미서+우미탁미정처리조(DEX+ AD조)、OLV+우미탁미정저제량조(DEX-L조)화OLV+우미탁미정고제량조(DEX-H조).Sham조대서행쌍폐통기2.5h.OLV조행우폐OLV 2.0h、쌍폐통기0.5h.DEX-L조화DEX-H조분별우OLV전1h정맥수주우미탁미정2.5 μg* kg-1·h-1、5.0μg·kg-1·h-1,1 h완성.AD조우OLV전1h정맥주사아체미서250 μg/kg.DEX+AD조우정맥수주DEX 5.0 μg·kg-1 h-1즉각정맥주사아체미서250 μg/kg.우쌍폐통기0.5h시처사대서취좌폐조직,측정폐조직습/간중비(W/D),광경하관찰폐조직병리개변병측정폐포손상솔(IAR),전경하관찰폐조직초미결구,TUNEL법검측폐조직세포조망,RT-PCR화Western blot법분별측정포도당조절단백78 (GRP78) mRNA급단백、JNK mRNA、린산화JNK (p-JNK)단백표체수평.결과 여Sham조비교,OLV조、AD조화DEX+AD조W/D、AI화IAR승고(P＜0.01);여OLV조、AD조화DEX+AD조비교,DEX-L조화DEX-H조W/D、AI화IAR강저(P＜0.01).OLV조、AD조화DEX+AD조폐조직결구균발생명현손상,이DEX-L조화DEX-H조폐조직결구손상칙명현감경.OLV조、AD조화DEX+AD조유대량폐혈관내피세포화폐포상피세포발생조망,이경DEX간예후세포조망칙명현감소.여Sham조비교,OLV조、AD조화DEX+AD조GRP78 mRNA화단백、JNK mRNA화p-JNK단백표체수평승고(P＜0.01);여OLV조、AD조화DEX+AD조비교,DEX-L조화DEX-H조GRP78 mRNA화단백、JNK mRNA화p-JNK단백표체수평강저(P＜0.01).결론 우미탁미정예선급약대단폐통기시적폐장구유보호작용,해작용적가능궤제여기억제JNK신호통로,감경ERS유도적세포조망유관.
Objective To evaluate the effects of dexmedetomidine (DEX) on cell apoptosis induced by endoplasmic reticulum stress and c-Jun N-terminal kinase (JNK) pathway during one-lung ventilation (OLV) in rats.Methods Sixty male Sprague-Dawley rats were randomly allocated into 6 groups (n =10 each):sham operation group (Sham group),OLV group,OLV + atipamezole (α2 receptor antagonist) group (AD group),OLV + atipamezole + DEX group (DEX+AD group),OLV + low-dose DEX group (DEX-L group) and OLV + high-dose DEX group (DEX-H group).The animals were anesthetized with 10％ chloral hydrate 4.5 ml/kg,tracheostomized and mechanically ventilated.Bilateral lungs were ventilated for 2.5 h in Sham group.The right lung was ventilated for 2.0 h followed by 0.5 h two-lung ventilation in OLV group.In DEX-L and DEX-H groups,DEX was infused intravenously for 1 h at a rate of 2.5 μg · kg-1 · h-1 and 5.0 μg · kg-1 · h-1,respectively,starting from 1 h prior to OLV.Atipamezole 250 μg/kg was injected intravenously at 1 h prior to OLV in AD group.Atipamezole 250 μg/kg was injected intravenously at the onset of DEX infusion (5.0 μg · kg-1 · h-1) in DEX+AD group.The rats were sacrificed and left lungs were removed for determination of weight to dry lung weight ratio (W/D),cell apoptosis in lung tissues (by TUNEL),and expression of glucose-regulated protein 78 (GRP78) mRNA and protein,JNK mRNA and phosphorylated JNK (p-JNK) protein (by RT-PCR and Western blot).Pathological changes of lungs were examined and the injured alveolus rate (IAR) was counted under light microscope.The changes in ultrastructure of lung tissues were observed under transmission electron microscope.Apoptosis index (AI) was calculated.Results W/D,AI and IAR were significantly higher in OLV,AD and DEX+AD group than in Sham group,while lower in DEX-L and DEX-H groups than in OLV,AD and DEX+AD groups.The pathological changes of the structure of lung tissues were observed in OLV,AD and DEX+AD groups,while the pathological changes were significantly alleviated in DEX-L and DEX-H groups.In OLV,AD and DEX + AD groups,there was apoptosis in lots of pulmonary vascular endothelial cells and alveolar epithelial cells,while cell apoptosis was significantly reduced after administration of DEX.The expression of GRP78 mRNA and protein,JNK mRNA and p-JNK protein was significantly higher in OLV,AD and DEX+AD groups than in Sham group,and lower in DEX-L and DEX-H groups than in OLV,AD and DEX +AD groups.Conclusion DEX pretreatment can protect lungs during OLV,and inhibited JNK signaling pathway and reduced cell apoptosis induced by endoplasmic reticulum stress may be involved in the mechanism.
