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
2011年
5期
375-379
,共5页
何白梅%罗百灵%彭振宇%袁浩
何白梅%囉百靈%彭振宇%袁浩
하백매%라백령%팽진우%원호
肺疾病,慢性阻塞性%内质网%细胞凋亡
肺疾病,慢性阻塞性%內質網%細胞凋亡
폐질병,만성조새성%내질망%세포조망
Pulmonary disease,chronic obstructive%Endoplasmic reticulum%Apoptosis
目的 观察COPD大鼠模型肺组织内质网应激的发生及肺泡上皮细胞凋亡.方法 24只Wistar雄性大鼠分为对照组和COPD模型组(简称模型组),每组12只.采用香烟烟雾暴露加气管内滴注脂多糖法建立COPD大鼠模型.造模完成后测定各组大鼠的肺功能,观察肺组织病理学变化,逆转录PCR法检测分子伴侣葡萄糖调节蛋白78(GRP78)、CCAAT/增强子结合蛋白同源蛋白(CHOP)mRNA水平,Western blot法检测GRP78、CHOP和活化的天冬氨酸特异性半胱氨酸蛋白酶-12(caspase-12)蛋白水平,原位缺口末端标记法检测大鼠肺泡上皮细胞凋亡.组间两两比较采用t检验.结果 模型组大鼠FEV0.3/FVC[(60±6)%]和动态肺顺应性[(0.17±0.02)cm H2O·ml-1·s-1,1 cm H2O=0.098 kPa]均明显低于对照组[(83±7)%和(0.31±0.03)cm H2O·ml-1·s-1],气道阻力[(0.64±0.07)ml/cm H2O]明显高于对照组[(0.32±0.03)ml/cm H2O],差异均有统计学意义(t值为-14.532～11.619,均P＜0.05);模型组大鼠支气管肺组织中GRP78和CHOP mRNA表达量的吸光度值(0.65±0.07和0.79±0.06)均明显高于对照组(0.21±0.04和0.07±0.04),差异均有统计学意义(t值分别为-19.102和-32.573,均P＜0.05);模型组大鼠肺组织中GRP78、CHOP和活化的caspase-12蛋白的吸光度值(0.83±0.06、0.82±0.06和0.81±0.07)均明显高于对照组(0.33±0.05、0.05±0.03和0.24±0.06),差异均有统计学意义(t值为-40.866～-22.070,均P＜0.05);模型组大鼠细胞凋亡指数[(32.4±3.7)%]明显高于对照组[(6.2±0.9)%],差异有统计学意义(t=-23.852,P＜0.05).结论 COPD大鼠模型的肺组织发生内质网应激,肺泡上皮细胞凋亡增加.内质网应激可能是介导肺泡上皮细胞凋亡的重要途径.
目的 觀察COPD大鼠模型肺組織內質網應激的髮生及肺泡上皮細胞凋亡.方法 24隻Wistar雄性大鼠分為對照組和COPD模型組(簡稱模型組),每組12隻.採用香煙煙霧暴露加氣管內滴註脂多糖法建立COPD大鼠模型.造模完成後測定各組大鼠的肺功能,觀察肺組織病理學變化,逆轉錄PCR法檢測分子伴侶葡萄糖調節蛋白78(GRP78)、CCAAT/增彊子結閤蛋白同源蛋白(CHOP)mRNA水平,Western blot法檢測GRP78、CHOP和活化的天鼕氨痠特異性半胱氨痠蛋白酶-12(caspase-12)蛋白水平,原位缺口末耑標記法檢測大鼠肺泡上皮細胞凋亡.組間兩兩比較採用t檢驗.結果 模型組大鼠FEV0.3/FVC[(60±6)%]和動態肺順應性[(0.17±0.02)cm H2O·ml-1·s-1,1 cm H2O=0.098 kPa]均明顯低于對照組[(83±7)%和(0.31±0.03)cm H2O·ml-1·s-1],氣道阻力[(0.64±0.07)ml/cm H2O]明顯高于對照組[(0.32±0.03)ml/cm H2O],差異均有統計學意義(t值為-14.532～11.619,均P＜0.05);模型組大鼠支氣管肺組織中GRP78和CHOP mRNA錶達量的吸光度值(0.65±0.07和0.79±0.06)均明顯高于對照組(0.21±0.04和0.07±0.04),差異均有統計學意義(t值分彆為-19.102和-32.573,均P＜0.05);模型組大鼠肺組織中GRP78、CHOP和活化的caspase-12蛋白的吸光度值(0.83±0.06、0.82±0.06和0.81±0.07)均明顯高于對照組(0.33±0.05、0.05±0.03和0.24±0.06),差異均有統計學意義(t值為-40.866～-22.070,均P＜0.05);模型組大鼠細胞凋亡指數[(32.4±3.7)%]明顯高于對照組[(6.2±0.9)%],差異有統計學意義(t=-23.852,P＜0.05).結論 COPD大鼠模型的肺組織髮生內質網應激,肺泡上皮細胞凋亡增加.內質網應激可能是介導肺泡上皮細胞凋亡的重要途徑.
목적 관찰COPD대서모형폐조직내질망응격적발생급폐포상피세포조망.방법 24지Wistar웅성대서분위대조조화COPD모형조(간칭모형조),매조12지.채용향연연무폭로가기관내적주지다당법건립COPD대서모형.조모완성후측정각조대서적폐공능,관찰폐조직병이학변화,역전록PCR법검측분자반려포도당조절단백78(GRP78)、CCAAT/증강자결합단백동원단백(CHOP)mRNA수평,Western blot법검측GRP78、CHOP화활화적천동안산특이성반광안산단백매-12(caspase-12)단백수평,원위결구말단표기법검측대서폐포상피세포조망.조간량량비교채용t검험.결과 모형조대서FEV0.3/FVC[(60±6)%]화동태폐순응성[(0.17±0.02)cm H2O·ml-1·s-1,1 cm H2O=0.098 kPa]균명현저우대조조[(83±7)%화(0.31±0.03)cm H2O·ml-1·s-1],기도조력[(0.64±0.07)ml/cm H2O]명현고우대조조[(0.32±0.03)ml/cm H2O],차이균유통계학의의(t치위-14.532～11.619,균P＜0.05);모형조대서지기관폐조직중GRP78화CHOP mRNA표체량적흡광도치(0.65±0.07화0.79±0.06)균명현고우대조조(0.21±0.04화0.07±0.04),차이균유통계학의의(t치분별위-19.102화-32.573,균P＜0.05);모형조대서폐조직중GRP78、CHOP화활화적caspase-12단백적흡광도치(0.83±0.06、0.82±0.06화0.81±0.07)균명현고우대조조(0.33±0.05、0.05±0.03화0.24±0.06),차이균유통계학의의(t치위-40.866～-22.070,균P＜0.05);모형조대서세포조망지수[(32.4±3.7)%]명현고우대조조[(6.2±0.9)%],차이유통계학의의(t=-23.852,P＜0.05).결론 COPD대서모형적폐조직발생내질망응격,폐포상피세포조망증가.내질망응격가능시개도폐포상피세포조망적중요도경.
Objective To study the endoplasmic retieulum stress(ERS)and the apoptosis of alveolar epithelial cells in a COPD rat model.Methods Twenty-four Wister rats were divided into a control group and a COPD group at random.The COPD rat model was established by intratracheal instillation of lipopolysaccharide(LPS)twice and exposure to cigarette smoke daily.The spirometry was conducted and the pathoiogical changes were observed after the model was established.The levels of glucose regulated protein 78(GRP78)and CCAAT/enhancer binding protein homologous protein(CHOP)mRNA were detected by reverse transcription-polymerase chain reaction(RT-PCR).The protein expression of GRP78,CHOP and caspase-12 was detected by Western blot.TdT-mediated dUTP nick end labeling(TUNEL)was used to analyze alveolar epithelial cell apoptosis.Comparisons between the two groups were performed by t-test. Results Significant decrease of FEV0.3/FVC[(60±6)%]and dynamic compliance of lung(CLdyn) [(0.17±0.02)cm H2O·ml-1·s-1],and increase of airway resistance[(0.64±0.07)ml/cm H2O]were found in the COPD group compared with the control group[(83±7)%,(0.31±0.03)cm H2O·ml-1·s-1,(0.32±0.03)ml/cm H2O](t=-14.532-11.619,P＜0.05).GRP78 mRNA and CHOP mRNA densitometry[(0.65±0.07),(0.79±0.06)] were significantly increased in the COPD group compared with the control group[(0.21±0.04),(0.07±0.04),respectively](t=-19.102 and -32.573,P＜0.05).GRP78,CHOP,and active caspase-12 protein densitometry(0.83±0.06,0.82±0.06 and 0.81±0.07)were significantly increased in the COPD group compared with the control group [(0.33±0.05,0.05±0.03 and 0.24±0.06),respectively](t=-40.866--22.070,P＜0.05).More apoptotic alveolar epithelial cells were found in the COPD group [(32.4±3.7)%] than in the control group [(6.2±0.9)%](t=-23.852,P＜0.05).Conclusions ERS was triggered in the lung tissues of COPD rats,especially in the alveolar epithelial cells.Alveolar epithelial cell apoptosis was increased in the COPD group.The ERS mediated apoptosis pathway may participate in the alveolar epithelial cell apoptosis in COPD.
