中华危重病急救医学
中華危重病急救醫學
중화위중병급구의학
Chinese Critical Care Medicine
2014年
6期
383-387
,共5页
张志坚%董瑶瑶%李晓萍%彭礼波
張誌堅%董瑤瑤%李曉萍%彭禮波
장지견%동요요%리효평%팽례파
中毒,百草枯%肺损伤%沙苑子总黄酮%c-Jun氨基末端激酶%内质网应激%葡萄糖调节蛋白78
中毒,百草枯%肺損傷%沙苑子總黃酮%c-Jun氨基末耑激酶%內質網應激%葡萄糖調節蛋白78
중독,백초고%폐손상%사원자총황동%c-Jun안기말단격매%내질망응격%포도당조절단백78
Paraquat poisoning%Lung injury%Total flavonoids from astragalus complanatus%c-Jun N-terminal kinase%Endoplasmic reticulum stress%Glucose regulated protein 78
目的 探讨沙苑子总黄酮(FAC)是否可通过抑制内质网应激和c-Jun氨基末端激酶(JNK)通路过度活化来减轻大鼠百草枯诱导的肺损伤.方法 48只SD大鼠按随机数字表法分为对照组、模型组、二甲基亚砜(DMSO)对照组和FAC低、中、高剂量组6组,每组8只.模型组采用PQ 80 mg/kg灌胃诱导肺损伤;对照组则给予等量生理盐水灌胃;DMSO组于PQ灌胃前2h腹腔注射10％ DMSO 20 mL/kg;FAC低、中、高剂量组于PQ灌胃后腹腔注射40、80、160 mg· kg-1· d-1的FAC溶液.制模后72 h处死大鼠取肺,检测肺湿/干质量(W/D)比值、总肺水含量(TLW);光镜下观察肺组织病理学改变,并进行肺组织损伤评估;逆转录-聚合酶链反应(RT-PCR)检测肺组织JNK和葡萄糖调节蛋白78(GRP78)的mRNA表达;蛋白质免疫印迹试验(Western Blot)检测肺组织JNK、磷酸化JNK(p-JNK)和GRP78的蛋白表达.结果 与对照组比较,模型组和DMSO组肺W/D比值、TLW、肺泡损伤定量评估指数(IQA)均明显升高,JNK、GRP78的mRNA表达和JNK、p-JNK、GRP78的蛋白表达均明显升高.与模型组比较,FAC各剂量组肺W/D比值、TLW、IQA以及JNK mRNA表达和p-JNK蛋白表达均明显下降,其中以FAC高剂量组降低最为显著[肺W/D比值:3.0±0.3比5.5±0.5,TLW:2.2±0.3比4.7±0.4,IQA:(15.4±3.0)％比(40.0±5.7)％,JNK mRNA:0.21 ±0.08比0.82±0.27,p-JNK蛋白:0.31±0.09比0.78±0.25,均P＜0.01];FAC低、中、高剂量组GRP78 mRNA和JNK、GRP78蛋白均为高表达,与模型组比较差异均无统计学意义(GRP78 mRNA:0.54±0.18比0.74±0.20,JNK蛋白:0.76±0.27比0.80±0.28,GRP78蛋白:0.51 ±0.18比0.69±0.21,均P＞0.05).结论 百草枯中毒后肺组织发生了过度的内质网应激损伤;FAC可能通过抑制内质网应激及JNK信号通路过度活化起到保护肺组织的作用.
目的 探討沙苑子總黃酮(FAC)是否可通過抑製內質網應激和c-Jun氨基末耑激酶(JNK)通路過度活化來減輕大鼠百草枯誘導的肺損傷.方法 48隻SD大鼠按隨機數字錶法分為對照組、模型組、二甲基亞砜(DMSO)對照組和FAC低、中、高劑量組6組,每組8隻.模型組採用PQ 80 mg/kg灌胃誘導肺損傷;對照組則給予等量生理鹽水灌胃;DMSO組于PQ灌胃前2h腹腔註射10％ DMSO 20 mL/kg;FAC低、中、高劑量組于PQ灌胃後腹腔註射40、80、160 mg· kg-1· d-1的FAC溶液.製模後72 h處死大鼠取肺,檢測肺濕/榦質量(W/D)比值、總肺水含量(TLW);光鏡下觀察肺組織病理學改變,併進行肺組織損傷評估;逆轉錄-聚閤酶鏈反應(RT-PCR)檢測肺組織JNK和葡萄糖調節蛋白78(GRP78)的mRNA錶達;蛋白質免疫印跡試驗(Western Blot)檢測肺組織JNK、燐痠化JNK(p-JNK)和GRP78的蛋白錶達.結果 與對照組比較,模型組和DMSO組肺W/D比值、TLW、肺泡損傷定量評估指數(IQA)均明顯升高,JNK、GRP78的mRNA錶達和JNK、p-JNK、GRP78的蛋白錶達均明顯升高.與模型組比較,FAC各劑量組肺W/D比值、TLW、IQA以及JNK mRNA錶達和p-JNK蛋白錶達均明顯下降,其中以FAC高劑量組降低最為顯著[肺W/D比值:3.0±0.3比5.5±0.5,TLW:2.2±0.3比4.7±0.4,IQA:(15.4±3.0)％比(40.0±5.7)％,JNK mRNA:0.21 ±0.08比0.82±0.27,p-JNK蛋白:0.31±0.09比0.78±0.25,均P＜0.01];FAC低、中、高劑量組GRP78 mRNA和JNK、GRP78蛋白均為高錶達,與模型組比較差異均無統計學意義(GRP78 mRNA:0.54±0.18比0.74±0.20,JNK蛋白:0.76±0.27比0.80±0.28,GRP78蛋白:0.51 ±0.18比0.69±0.21,均P＞0.05).結論 百草枯中毒後肺組織髮生瞭過度的內質網應激損傷;FAC可能通過抑製內質網應激及JNK信號通路過度活化起到保護肺組織的作用.
목적 탐토사원자총황동(FAC)시부가통과억제내질망응격화c-Jun안기말단격매(JNK)통로과도활화래감경대서백초고유도적폐손상.방법 48지SD대서안수궤수자표법분위대조조、모형조、이갑기아풍(DMSO)대조조화FAC저、중、고제량조6조,매조8지.모형조채용PQ 80 mg/kg관위유도폐손상;대조조칙급여등량생리염수관위;DMSO조우PQ관위전2h복강주사10％ DMSO 20 mL/kg;FAC저、중、고제량조우PQ관위후복강주사40、80、160 mg· kg-1· d-1적FAC용액.제모후72 h처사대서취폐,검측폐습/간질량(W/D)비치、총폐수함량(TLW);광경하관찰폐조직병이학개변,병진행폐조직손상평고;역전록-취합매련반응(RT-PCR)검측폐조직JNK화포도당조절단백78(GRP78)적mRNA표체;단백질면역인적시험(Western Blot)검측폐조직JNK、린산화JNK(p-JNK)화GRP78적단백표체.결과 여대조조비교,모형조화DMSO조폐W/D비치、TLW、폐포손상정량평고지수(IQA)균명현승고,JNK、GRP78적mRNA표체화JNK、p-JNK、GRP78적단백표체균명현승고.여모형조비교,FAC각제량조폐W/D비치、TLW、IQA이급JNK mRNA표체화p-JNK단백표체균명현하강,기중이FAC고제량조강저최위현저[폐W/D비치:3.0±0.3비5.5±0.5,TLW:2.2±0.3비4.7±0.4,IQA:(15.4±3.0)％비(40.0±5.7)％,JNK mRNA:0.21 ±0.08비0.82±0.27,p-JNK단백:0.31±0.09비0.78±0.25,균P＜0.01];FAC저、중、고제량조GRP78 mRNA화JNK、GRP78단백균위고표체,여모형조비교차이균무통계학의의(GRP78 mRNA:0.54±0.18비0.74±0.20,JNK단백:0.76±0.27비0.80±0.28,GRP78단백:0.51 ±0.18비0.69±0.21,균P＞0.05).결론 백초고중독후폐조직발생료과도적내질망응격손상;FAC가능통과억제내질망응격급JNK신호통로과도활화기도보호폐조직적작용.
Objective To investigate the effect of total flavonoids from astragalus complanatus (FAC) on attenuating lung injury resulted from paraquat (PQ) poisoning by inhibiting excessive endoplasmic reticulum stress (ERS) and c-Jun N-terminal kinase (JNK) pathway in rat.Methods Forty-eight Sprague-Dawley (SD) rats were randomly divided into six groups (n=8 in each group),including control group,model group,dimethyl sulfoxide (DMSO) vehicle control group,and FAC in low,medium,and high dosage groups.The model was reproduced by giving PQ 80 mg/kg orally to induce lung injury.The rats in control group were treated with saline by gavage.The rats in DMSO group were given 10％ DMSO 20 mL/kg by gavage 2 hours before intraperitoneal injection of PQ,and those in FAC low,medium and high dosage groups received 40,80,160 mg·kg-1· d-1 of FAC solution intraperitoneally after the PQ administration.The rats were sacrificed 72 hours after giving PQ,and the left lung tissue was harvested 72 hours after the reproduction of experimental model.The ratio of wet/dry weight (W/D) and total lung water content (TLW) were determined.The pathohistological changes of the left lung was observed under light microscope,and scored with alveolar damage index of quantitative assessment (IQA).The mRNA expressions of JNK and glucose regulated protein 78 (GRP78) were determined by reverse transcription-polymerase chain reaction (RT-PCR),and the protein expression of JNK,phosphorylation-JNK (p-JNK),and GRP78 were determined by Western Blot.Results Compared with control group,the W/D ratio,TLW and IQA were increased significantly in model group and DMSO group,and the mRNA expressions of JNK and GRP78 and the protein expressions of JNK,p-JNK and GRP78 were markedly increased.Compared with the model group,the W/D ratio,TLW and IQA,and the expressions of JNK mRNA and p-JNK protein were significantly decreased in the FAC groups,especially in FAC high dosage group [W/D ratio:3.0 ± 0.3 vs.5.5 ± 0.5,TLW:2.2 ± 0.3 vs.4.7 ± 0.4,IQA:(15.4 ± 3.0)％ vs.(40.0 ± 5.7)％,JNK mRNA:0.21 ± 0.08 vs.0.82 ±0.27,p-JNK protein:0.31 ±0.09 vs.0.78 ±0.25,all P＜0.O1].The mRNA expression of GRP78 and the protein expressions of JNK and GRP78 were highly expressed in FAC low,medium and high dosage groups,and there was no significant difference compared with those in model group (GRP78 mRNA:0.54 ± 0.18 vs.0.74 ± 0.20,JNK protein:0.76 ± 0.27 vs.0.80 ± 0.28,GRP78 protein:0.51 ± 0.18 vs.0.69 ± 0.21,all P＞0.05).Conclusions PQ induces excessive ERS in the lung tissue resulting in lung injury.FAC has a protective effect on lung against PQ injury,and it may be related with inhibition JNK pathway in ERS.
