中华结核和呼吸杂志
中華結覈和呼吸雜誌
중화결핵화호흡잡지
Chinese Journal of Tuberculosis and Respiratory Diseases
2014年
1期
30-35
,共6页
李玉屏%罗百灵%张立%马苗%郭现玲
李玉屏%囉百靈%張立%馬苗%郭現玲
리옥병%라백령%장립%마묘%곽현령
肺疾病,慢性阻塞性%藜芦属%细胞凋亡%上皮细胞
肺疾病,慢性阻塞性%藜蘆屬%細胞凋亡%上皮細胞
폐질병,만성조새성%려호속%세포조망%상피세포
Pulmonary disease,chronic obstructive%Veratrum%Apoptosis%Epithelial cells
目的 探索白藜芦醇对减轻慢性阻塞性肺疾病(简称慢阻肺)大鼠内质网应激诱导肺泡上皮细胞凋亡的保护作用.方法 将48只雄性SD大鼠分为正常对照组、白藜芦醇对照组、慢阻肺组和白藜芦醇干预组,每组12只.慢阻肺组和白藜芦醇干预组采用反复烟草烟雾暴露联合气管内注入脂多糖法制备大鼠慢阻肺模型,白藜芦醇对照组和白藜芦醇干预组大鼠采用白藜芦醇25 mg·kg-1·d-1灌胃.造模后进行肺功能、肺组织病理学、免疫组织化学和Western blot检测,观察肺组织内质网应激凋亡分子CCAAT/增强子结合蛋白同源蛋白(CHOP)和天冬氨酸特异性半胱氨酸蛋白酶-12(caspase-12)蛋白的表达.采用原位末端标记法检测细胞凋亡.多组间比较采用单因素方差分析,组间两两比较采用LSD-t检验.结果 慢阻肺组大鼠FEV03/FVC[(59±4)％]和动态肺顺应性[(0.154 ±0.013) ml/cmH2O,1 cmH2O =0.098 kPa]显著低于正常对照组[(82±4)％和(0.401±0.033) ml/cmH2O],气道阻力[(0.651 ±0.046) cmH2O·ml-1· s-1]显著高于正常对照组[(0.404±0.033) cmiH2O·ml-·s-](t值为-14.48 ～ 16.48,均P＜0.05)；白藜芦醇干预组大鼠FEV03/FVC[(71±5)％]和动态肺顺应性[(0.302±0.023) ml/cmH2O]显著高于慢阻肺组,气道阻力[(0.442±0.036) cmiH2O·ml-1·s-1]显著低于慢阻肺组(t值为-10.02 ～ 10.37,均P＜0.05).慢阻肺组大鼠肺组织中可见明显的支气管炎症和肺气肿改变,较白藜芦醇干预组的程度重；肺组织中CHOP和caspase-12的积分吸光度值分别为9 778 ±217和12 009±346,显著高于正常对照组(960±94和1 124±112),白藜芦醇干预组(5 799±177和6 720±173)显著低于慢阻肺组,差异均有统计学意义(t值为-100.43 ～ 45.32,均P＜0.05).Westem blot检测结果显示,慢阻肺组大鼠肺组织中CHOP和caspase-12相对表达量分别为0.910±0.053和1.104±0.026,显著高于正常对照组(0.204 ±0.021和0.133-±0.013),白藜芦醇干预组(0.462±0.037和0.642 ±0.011)显著低于慢阻肺组,差异均有统计学意义(t值为-115.03 ～ 60.59,均P＜0.05).慢阻肺组肺泡上皮细胞凋亡指数[(39.8±1.6)％]显著高于白藜芦醇干预组和正常对照组[(26.3±1.5)％和(6.4±0.6)％],差异均有统计学意义(t值分别为20.21和-49.94,均P＜0.05).结论 慢阻肺大鼠肺组织发生内质网应激,且诱导肺泡上皮细胞凋亡增加,白藜芦醇可减轻内质网应激诱导的肺泡上皮细胞凋亡.
目的 探索白藜蘆醇對減輕慢性阻塞性肺疾病(簡稱慢阻肺)大鼠內質網應激誘導肺泡上皮細胞凋亡的保護作用.方法 將48隻雄性SD大鼠分為正常對照組、白藜蘆醇對照組、慢阻肺組和白藜蘆醇榦預組,每組12隻.慢阻肺組和白藜蘆醇榦預組採用反複煙草煙霧暴露聯閤氣管內註入脂多糖法製備大鼠慢阻肺模型,白藜蘆醇對照組和白藜蘆醇榦預組大鼠採用白藜蘆醇25 mg·kg-1·d-1灌胃.造模後進行肺功能、肺組織病理學、免疫組織化學和Western blot檢測,觀察肺組織內質網應激凋亡分子CCAAT/增彊子結閤蛋白同源蛋白(CHOP)和天鼕氨痠特異性半胱氨痠蛋白酶-12(caspase-12)蛋白的錶達.採用原位末耑標記法檢測細胞凋亡.多組間比較採用單因素方差分析,組間兩兩比較採用LSD-t檢驗.結果 慢阻肺組大鼠FEV03/FVC[(59±4)％]和動態肺順應性[(0.154 ±0.013) ml/cmH2O,1 cmH2O =0.098 kPa]顯著低于正常對照組[(82±4)％和(0.401±0.033) ml/cmH2O],氣道阻力[(0.651 ±0.046) cmH2O·ml-1· s-1]顯著高于正常對照組[(0.404±0.033) cmiH2O·ml-·s-](t值為-14.48 ～ 16.48,均P＜0.05)；白藜蘆醇榦預組大鼠FEV03/FVC[(71±5)％]和動態肺順應性[(0.302±0.023) ml/cmH2O]顯著高于慢阻肺組,氣道阻力[(0.442±0.036) cmiH2O·ml-1·s-1]顯著低于慢阻肺組(t值為-10.02 ～ 10.37,均P＜0.05).慢阻肺組大鼠肺組織中可見明顯的支氣管炎癥和肺氣腫改變,較白藜蘆醇榦預組的程度重；肺組織中CHOP和caspase-12的積分吸光度值分彆為9 778 ±217和12 009±346,顯著高于正常對照組(960±94和1 124±112),白藜蘆醇榦預組(5 799±177和6 720±173)顯著低于慢阻肺組,差異均有統計學意義(t值為-100.43 ～ 45.32,均P＜0.05).Westem blot檢測結果顯示,慢阻肺組大鼠肺組織中CHOP和caspase-12相對錶達量分彆為0.910±0.053和1.104±0.026,顯著高于正常對照組(0.204 ±0.021和0.133-±0.013),白藜蘆醇榦預組(0.462±0.037和0.642 ±0.011)顯著低于慢阻肺組,差異均有統計學意義(t值為-115.03 ～ 60.59,均P＜0.05).慢阻肺組肺泡上皮細胞凋亡指數[(39.8±1.6)％]顯著高于白藜蘆醇榦預組和正常對照組[(26.3±1.5)％和(6.4±0.6)％],差異均有統計學意義(t值分彆為20.21和-49.94,均P＜0.05).結論 慢阻肺大鼠肺組織髮生內質網應激,且誘導肺泡上皮細胞凋亡增加,白藜蘆醇可減輕內質網應激誘導的肺泡上皮細胞凋亡.
목적 탐색백려호순대감경만성조새성폐질병(간칭만조폐)대서내질망응격유도폐포상피세포조망적보호작용.방법 장48지웅성SD대서분위정상대조조、백려호순대조조、만조폐조화백려호순간예조,매조12지.만조폐조화백려호순간예조채용반복연초연무폭로연합기관내주입지다당법제비대서만조폐모형,백려호순대조조화백려호순간예조대서채용백려호순25 mg·kg-1·d-1관위.조모후진행폐공능、폐조직병이학、면역조직화학화Western blot검측,관찰폐조직내질망응격조망분자CCAAT/증강자결합단백동원단백(CHOP)화천동안산특이성반광안산단백매-12(caspase-12)단백적표체.채용원위말단표기법검측세포조망.다조간비교채용단인소방차분석,조간량량비교채용LSD-t검험.결과 만조폐조대서FEV03/FVC[(59±4)％]화동태폐순응성[(0.154 ±0.013) ml/cmH2O,1 cmH2O =0.098 kPa]현저저우정상대조조[(82±4)％화(0.401±0.033) ml/cmH2O],기도조력[(0.651 ±0.046) cmH2O·ml-1· s-1]현저고우정상대조조[(0.404±0.033) cmiH2O·ml-·s-](t치위-14.48 ～ 16.48,균P＜0.05)；백려호순간예조대서FEV03/FVC[(71±5)％]화동태폐순응성[(0.302±0.023) ml/cmH2O]현저고우만조폐조,기도조력[(0.442±0.036) cmiH2O·ml-1·s-1]현저저우만조폐조(t치위-10.02 ～ 10.37,균P＜0.05).만조폐조대서폐조직중가견명현적지기관염증화폐기종개변,교백려호순간예조적정도중；폐조직중CHOP화caspase-12적적분흡광도치분별위9 778 ±217화12 009±346,현저고우정상대조조(960±94화1 124±112),백려호순간예조(5 799±177화6 720±173)현저저우만조폐조,차이균유통계학의의(t치위-100.43 ～ 45.32,균P＜0.05).Westem blot검측결과현시,만조폐조대서폐조직중CHOP화caspase-12상대표체량분별위0.910±0.053화1.104±0.026,현저고우정상대조조(0.204 ±0.021화0.133-±0.013),백려호순간예조(0.462±0.037화0.642 ±0.011)현저저우만조폐조,차이균유통계학의의(t치위-115.03 ～ 60.59,균P＜0.05).만조폐조폐포상피세포조망지수[(39.8±1.6)％]현저고우백려호순간예조화정상대조조[(26.3±1.5)％화(6.4±0.6)％],차이균유통계학의의(t치분별위20.21화-49.94,균P＜0.05).결론 만조폐대서폐조직발생내질망응격,차유도폐포상피세포조망증가,백려호순가감경내질망응격유도적폐포상피세포조망.
Objective To investigate alveolar epithelial cell apoptosis induced by endoplasmic reticulum stress in a rat model of chronic obstructive pulmonary disease (COPD) and the potential protective effect of resveratrol.Methods The COPD rat model was established by intratracheal instillation of lipopolysaccharide (LPS) and exposure to cigarette smoke daily.Forty-eight male Sprague-Dawley rats were randomly divided into 4 groups (n =12 each):a normal control group,a resveratrol control group (resveratrol 25 mg · kg-1 · d-1 gavage),a COPD group (COPD rat model established),and a resveratrol intervention group (COPD model rats receiving resveratrol 25 mg · kg-1 · d-1 gavage).Spirometry was conducted and the lung pathological changes were observed.The protein expression of CCAAT/enhancer binding protein homologous protein (CHOP) and caspase-12 were detected by immunohistochemistry and Western blot,and alveolar epithelial apoptosis was analyzed by terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL).Statistical analysis among groups were carried out by one way analysis of variance followed by LSD-t test between 2 groups.Results Significant decrease of FEV0.3/FVC [(59 ± 4) ％] and dynamic lung compliance [(0.154 ± 0.013) ml/cmH2 O,1 cmH2O =0.098 kPa] and increase of airway resistance [(0.651 ±0.046) cmH2O · ml-1 · s-1] were found in the COPD group when compared with the normal control group [(82 ± 4) ％,(0.401 ± 0.033) ml/cmH2O,(0.404-± 0.033)cmH2O·ml-1 ·s-1] (t =-14.48,16.48,P＜0.05).The FEV0.3/FVC [(71 ±5)％] and dynamic lung compliance [(0.302 ± 0.023) ml/cmH2O] of the resveratrol intervention group were significantly improved when compared with those of the COPD group,and the airway resistance [(0.442 ± 0.036) cmH2O · m1-1 · s-1] also decreased (t =-10.02-10.37,P ＜ 0.05).Significant small airway inflammation and emphysema were seen in the lung tissue of COPD group,while significant improvement was observed in the resveratrol intervention group when compared with COPD group.The lung tissue immunohistochemistry integrated optical density (IOD) of CHOP and caspase-12 (9 778 ± 217,12 009 ± 346) of the COPD group increased significantly when compared with the normal control group (960 ± 94,1 124 ± 112) (t =-100.43,-90.43,P ＜ 0.05),while the IODs of the resveratrol intervention group (5 799 ± 177,6 720 ± 173) decreased significantly when compared with the COPD group (t =45.32,43.93,P ＜0.05).Western blot results showed that the relative quantification of CHOP (0.910 ±0.053)and caspase-12 (1.104 ± 0.026) increased in the COPD group when compared with the normal control group (0.204 ±0.021,0.133 ±0.013,t =-36.04,-115.03,P ＜0.05),while the ratios of the resveratrol intervention group (0.462 ± 0.037,0.642 ± 0.011) decreased significantly when compared with COPD group (t =24.22,60.59,P ＜ 0.05).Higher apoptosis index was seen in the COPD group [(39.8 ±1.6) ％] when compared with the resveratrol intervention group [(26.3 ± 1.5) ％] and the normal control group [(6.4 ± 0.6) ％] (t =20.21,-49.94,P ＜ 0.05).Conclusions Endoplasmic reticulum stress,which induced apoptosis of alveolar epithelial cells,was observed in this COPD model.Resveratrol was shown to alleviate endoplasmic reticulum stress and attenuate alveolar epithelial apoptosis.
