中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2015年
18期
1415-1420
,共6页
陈燕%余常辉%潘妙霞%李婷%李维%蔡绍曦%孟莹
陳燕%餘常輝%潘妙霞%李婷%李維%蔡紹晞%孟瑩
진연%여상휘%반묘하%리정%리유%채소희%맹형
基质金属蛋白酶9%大鼠,近交SHR%肺损伤%烟熏
基質金屬蛋白酶9%大鼠,近交SHR%肺損傷%煙熏
기질금속단백매9%대서,근교SHR%폐손상%연훈
Matrix metalloproteinase 9%Rats,inbred SHR%Pulmonary injury%Cigarette smoking
目的 比较基质金属蛋白酶9(MMP9)在Wistar大鼠和自发性高血压大鼠烟熏肺损伤中的变化.方法 雄性Wistar大鼠及雄性自发性高血压(SH)大鼠各10只,分别按随机数字表法随机分为Wistar对照组、Wistar烟熏组及SH对照组、SH烟熏组各5只.对照组常规饲养,烟熏组置于自制有机玻璃箱内,每天烟熏2次,每周烟熏6d.8周后观察大鼠一般情况,检测其肺功能及肺组织病理学;Western印迹法检测各组肺组织中核因子(NF)-κB、IκB-α、MMP9蛋白表达;实时定量PCR检测肿瘤坏死因子(TNF)-α和白细胞介素(IL)-6 mRNA表达的变化,免疫组化检测各组肺组织中MMP9蛋白的表达.结果 两烟熏组大鼠出现消瘦,间歇咳嗽及气促.Wistar对照组、Wistar烟熏组及SH对照组、SH烟熏组气道阻力分别为(0.23±0.04)、(0.33 ±0.05)、(0.33±0.02)、(0.46±0.08)cmH2O ·s· ml-1,Wistar烟熏组显著高于Wistar对照组,SH烟熏组显著高于SH对照组和Wistar烟熏组(均P＜0.05).肺组织HE染显示两烟熏组大鼠均有慢性支气管炎和肺气肿的改变,SH烟熏组病变更为明显.两烟熏组平均肺泡数均显著低于其对照组(均P＜0.05),Wistar烟熏组平均内衬间隔、肺泡破坏指数、支气管平滑肌指数和胶原指数均显著高于Wistar对照组,SH烟熏组均显著高于SH对照组和Wistar烟熏组(均P＜0.05).Western印迹法检测Wistar对照组、Wistar烟熏组及SH对照组、SH烟熏组NF-κB蛋白表达分别为(0.322 ±0.014)、(0.558 ±0.044)、(0.373±0.029)、(1.156±0.197),MMP9蛋白表达分别为(0.255±0.070)、(0.456±0.089)、(0.594±0.184)、(0.847 ±0.138),SH烟熏组均显著高于SH对照组和Wistar烟熏组(均P＜0.05);SH烟熏组IκB-α蛋白表达均显著低于SH对照组和Wistar烟熏组(均P＜0.05).两烟熏组TNF-α mRNA表达水平均显著高于其对照组(均P＜0.05),SH烟熏组IL-6 mRNA表达水平均显著高于SH对照组和Wistar烟熏组(均P＜0.05).肺组织MMP9免疫组化结果显示烟熏组表达水平均明显高于对照组,SH烟熏组表达明显高于Wistar烟熏组(均P＜0.05).结论 Wistar大鼠与SH大鼠烟熏后肺部均出现类似人类慢性阻塞性肺疾病的损伤,SH大鼠比Wistar大鼠肺损伤表现更为严重,MMP9表达水平明显增加.
目的 比較基質金屬蛋白酶9(MMP9)在Wistar大鼠和自髮性高血壓大鼠煙熏肺損傷中的變化.方法 雄性Wistar大鼠及雄性自髮性高血壓(SH)大鼠各10隻,分彆按隨機數字錶法隨機分為Wistar對照組、Wistar煙熏組及SH對照組、SH煙熏組各5隻.對照組常規飼養,煙熏組置于自製有機玻璃箱內,每天煙熏2次,每週煙熏6d.8週後觀察大鼠一般情況,檢測其肺功能及肺組織病理學;Western印跡法檢測各組肺組織中覈因子(NF)-κB、IκB-α、MMP9蛋白錶達;實時定量PCR檢測腫瘤壞死因子(TNF)-α和白細胞介素(IL)-6 mRNA錶達的變化,免疫組化檢測各組肺組織中MMP9蛋白的錶達.結果 兩煙熏組大鼠齣現消瘦,間歇咳嗽及氣促.Wistar對照組、Wistar煙熏組及SH對照組、SH煙熏組氣道阻力分彆為(0.23±0.04)、(0.33 ±0.05)、(0.33±0.02)、(0.46±0.08)cmH2O ·s· ml-1,Wistar煙熏組顯著高于Wistar對照組,SH煙熏組顯著高于SH對照組和Wistar煙熏組(均P＜0.05).肺組織HE染顯示兩煙熏組大鼠均有慢性支氣管炎和肺氣腫的改變,SH煙熏組病變更為明顯.兩煙熏組平均肺泡數均顯著低于其對照組(均P＜0.05),Wistar煙熏組平均內襯間隔、肺泡破壞指數、支氣管平滑肌指數和膠原指數均顯著高于Wistar對照組,SH煙熏組均顯著高于SH對照組和Wistar煙熏組(均P＜0.05).Western印跡法檢測Wistar對照組、Wistar煙熏組及SH對照組、SH煙熏組NF-κB蛋白錶達分彆為(0.322 ±0.014)、(0.558 ±0.044)、(0.373±0.029)、(1.156±0.197),MMP9蛋白錶達分彆為(0.255±0.070)、(0.456±0.089)、(0.594±0.184)、(0.847 ±0.138),SH煙熏組均顯著高于SH對照組和Wistar煙熏組(均P＜0.05);SH煙熏組IκB-α蛋白錶達均顯著低于SH對照組和Wistar煙熏組(均P＜0.05).兩煙熏組TNF-α mRNA錶達水平均顯著高于其對照組(均P＜0.05),SH煙熏組IL-6 mRNA錶達水平均顯著高于SH對照組和Wistar煙熏組(均P＜0.05).肺組織MMP9免疫組化結果顯示煙熏組錶達水平均明顯高于對照組,SH煙熏組錶達明顯高于Wistar煙熏組(均P＜0.05).結論 Wistar大鼠與SH大鼠煙熏後肺部均齣現類似人類慢性阻塞性肺疾病的損傷,SH大鼠比Wistar大鼠肺損傷錶現更為嚴重,MMP9錶達水平明顯增加.
목적 비교기질금속단백매9(MMP9)재Wistar대서화자발성고혈압대서연훈폐손상중적변화.방법 웅성Wistar대서급웅성자발성고혈압(SH)대서각10지,분별안수궤수자표법수궤분위Wistar대조조、Wistar연훈조급SH대조조、SH연훈조각5지.대조조상규사양,연훈조치우자제유궤파리상내,매천연훈2차,매주연훈6d.8주후관찰대서일반정황,검측기폐공능급폐조직병이학;Western인적법검측각조폐조직중핵인자(NF)-κB、IκB-α、MMP9단백표체;실시정량PCR검측종류배사인자(TNF)-α화백세포개소(IL)-6 mRNA표체적변화,면역조화검측각조폐조직중MMP9단백적표체.결과 량연훈조대서출현소수,간헐해수급기촉.Wistar대조조、Wistar연훈조급SH대조조、SH연훈조기도조력분별위(0.23±0.04)、(0.33 ±0.05)、(0.33±0.02)、(0.46±0.08)cmH2O ·s· ml-1,Wistar연훈조현저고우Wistar대조조,SH연훈조현저고우SH대조조화Wistar연훈조(균P＜0.05).폐조직HE염현시량연훈조대서균유만성지기관염화폐기종적개변,SH연훈조병변경위명현.량연훈조평균폐포수균현저저우기대조조(균P＜0.05),Wistar연훈조평균내츤간격、폐포파배지수、지기관평활기지수화효원지수균현저고우Wistar대조조,SH연훈조균현저고우SH대조조화Wistar연훈조(균P＜0.05).Western인적법검측Wistar대조조、Wistar연훈조급SH대조조、SH연훈조NF-κB단백표체분별위(0.322 ±0.014)、(0.558 ±0.044)、(0.373±0.029)、(1.156±0.197),MMP9단백표체분별위(0.255±0.070)、(0.456±0.089)、(0.594±0.184)、(0.847 ±0.138),SH연훈조균현저고우SH대조조화Wistar연훈조(균P＜0.05);SH연훈조IκB-α단백표체균현저저우SH대조조화Wistar연훈조(균P＜0.05).량연훈조TNF-α mRNA표체수평균현저고우기대조조(균P＜0.05),SH연훈조IL-6 mRNA표체수평균현저고우SH대조조화Wistar연훈조(균P＜0.05).폐조직MMP9면역조화결과현시연훈조표체수평균명현고우대조조,SH연훈조표체명현고우Wistar연훈조(균P＜0.05).결론 Wistar대서여SH대서연훈후폐부균출현유사인류만성조새성폐질병적손상,SH대서비Wistar대서폐손상표현경위엄중,MMP9표체수평명현증가.
Objective To compare the expression of matrix metalloproteinase-9 (MMP9) and observe the change of cigarette smoke-induced pulmonary injury in Wistar and spontaneously hypertensive (SH) rats.Methods A total of 10 male Wistar rats and 10 male SH rats were randomly divided into four groups of Wistar control,Wistar cigarette smoking,SH control and SH cigarette smoking (n =5 each).The control groups were fed routinely while the cigarette smoking groups were placed into a homemade organic glass box and exposed to cigarette smoking for twice daily 6 days a week.After 8-week treatment,general condition,lung function and pathological changes of lung tissues were detected.The expressions of nuclear factor (NF)-κB,IκB-α and MMP9 were determined by Western blot.And real-time polymerase chain reaction (RT-PCR) was employed to detect the mRNA levels of tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6).Immunohistochemistry was used to detect the expression of MMP9 in lung tissue.Results Both cigarette smoking groups were marantic along with intermittent cough and polypnea.Resistance index (RI) in Wistar control,Wistar cigarette smoking,SH control and SH cigarette smoking groups was (0.23 ±0.04),(0.33 ±0.05),(0.33 ±0.02) and (0.46 ±0.08) cmH2O · s · ml-1 respectively.RI increased in Wistar cigarette smoking group versus Wistar control group.And SH cigarette smoking group was significantly higher than SH control and Wistar cigarette smoking groups (P ＜ 0.05).Hematoxylin & eosin staining demonstrated that the lesion of chronic bronchitic inflammation and emphysema existed in two cigarette smoking groups while it was more severe in SH rats than Wistar rats.The mean alveolar number (MAN) was significantly reduced in two cigarette smoking groups compared with that in control groups (P ＜ 0.05,respectively).And mean lining interval (MLI),damage index (DI),bronchial smooth muscle index and collagen index significantly increased in Wistar cigarette smoking group compared with Wistar control group,SH cigarette smoking group significantly increased than that in SH control and Wistar cigarette smoking groups (P ＜ 0.05,respectively).On Western blot,the protein level of NF-κB in Wistar control,Wistar cigarette smoking,SH control and SH cigarette smoking groups was (0.322 ± 0.014),(0.558 ± 0.044),(0.373 ± 0.029) and (1.156 ± 0.197) respectively.The protein level of MMP9 in these groups was (0.255 ±0.070),(0.456 ±0.089),(0.594 ±0.184) and (0.847 ±0.138)respectively.The protein levels of NF-κB and MMP9 increased obviously in Wistar cigarette smoking group versus Wistar control group and those in SH cigarette smoking group were significantly higher than SH control and Wistar cigarette smoking groups (P ＜ 0.05).Additionally,IκB-α expression in SH cigarette smoking group was markedly decreased in comparison to that in SH control group and Wistar cigarette smoking group (P ＜ 0.05).The mRNA level of TNF-α increased obviously in two cigarette smoking groups in comparison to those in controls (P ＜ 0.05) and the mRNA level of IL-6 significantly increased in the SH cigarette smoking group compared with SH control group and Wistar cigarette smoking group (P ＜ 0.05).As for immunohistochemistry,the MMP9 positive cells were prominently distributed in the lungs of cigarette smoking groups versus the stained cells in the control group.And SH cigarette smoking group was significantly higher than Wistar cigarette smoking group (P ＜ 0.05).Conclusion Both Wistar and SH rats induced by cigarette smoking may have an onset of lung injury symptom analogous to chronic obstructive pulmonary disease and SH rat are more severe.
