肺气肿合并睡眠诱导低氧大鼠模型的建立及凝血功能的变化

目的 创新建立肺气肿合并睡眠诱导低氧大鼠模型并探讨凝血功能的变化.方法 纳入30只Wistar大鼠,按暴露条件分3组.每组10只.A组(单纯睡眠诱导低氧组).正常饲养28 d,第29天起在睡眠时段暴露于含12.5%氧的混合气中共3 h(分4次给予,每次45 min);B组(单纯肺气肿组),每天分两次暴露于纸烟烟雾共约1 h(每次约30 min);C组(肺气肿合并睡眠诱导低氧组),前28 d同B组暴露,第29天起继续予熏烟暴露,同时在睡眠时段暴露于含12.5%氧的混合气中共3 h(分4次给予,每次45 min).各组大鼠连续暴露至56 d后处死,采集肺组织和全血.肺组织切片染色后经半定量图像分析,计算肺病理评分、肺平均内衬间隔(MLI)和平均肺泡数(MAN)来评价肺组织病理学改变.而后测定全血中抗凝血酶Ⅲ(AT Ⅲ)、纤维蛋白原(FIB)、血管性血友病因子(vWF)、凝血因子VⅢ(FⅧ)的活性.以SPSS 11.5进行统计学处理.采用单因素方差分析行组间比较,组内比较采用SNK-q检验.结果 B、C组大鼠肺组织切片均呈现明显肺气肿样改变.各组肺病理评分和MLI差异明显(F=520.751、F＝1 037.787,P值均<0.05).B组[(52.00±5.16)%、(91.30+2.26)μm]和C组[(73.30±3.86)%、(99.32±2.81)μm]均高于A组[(16.20±2.53)%、(46.79±3.18)μm,q=28.312、q＝45.157、q＝50.666、q=59.800,P值均<0.053,且C组高于B组(q=16.845、q=9.135,P值均<0.05);各组MAN差异明显(F＝313.304,P<0.05),B组[(103.55±4.45)/mm2]和C组[(83.70±4.72)/mm2]均低于A组[(151.59±8.64)/mm2,q=24.358、q=34.426,P值均<0.053],且C组低于B组(q=10.068.P<0.05).各组大鼠全血ATⅢ活性差异明显(F＝25.386.P<0.05).B组[(107.39±8.05)%]和C组[(99.04±3.99)%]ATⅢ活性均低于A组[(116.20±2.53)%.q＝5.173、q=10.076,P值均<0.05].且C组低于B组(q=4.903,P<0.05);各组大鼠全血FIB、vWF和FⅧ活性差异明显(F=112.969、F=223.666,F＝70.040.P值均<0.05),B组[(1774.60±72.49)mg/L、(64.45±1.72)%、(227.78±10.38)%]和C组[(1984.00c73.69)mg/L、(71.96±1.85)%、(303.25±32.93)%]FIB均高于A组[(1515.50±62.79)mg/L、(56.02±1.47)%、(200.36±4.88)%,q=11.734、q=21.218、q=15.810、q=29.895,q=4.307、q=16.16l,P值均<0.05],且C组高于B组(q=9.483,q＝14.085、q=11.854.P值均<0.05).结论 本研究成功建立了肺气肿合并睡眠诱导低氧大鼠模型.肺组织切片病理提示肺气肿大鼠合并睡眠诱导低氧后肺气肿程度加重;大鼠患肺气肿后血液凝固性增强,而合并睡眠诱导低氧后,该血液高凝状态进一步加重.

Model of obstructive pulmonary emphysema with sleep-induced hypoxemia and function changes of coagulation system

Objective To establish a model in rats of pulmonary emphysema with sleep-induced hypoxemia(SIH),and discuss the function changes of coagulation system.Methods According to the different exposure condition,30 Wistar rats were randomly and averagely divided into three groups, 10 each group. Group A (the group of SIH) ,the rats were bred normally for 28 days,then exposed to the mixed gas of 12.5% oxygen for three hours during sleeping time every day (the exposed time was divided into four periods,45 rain each period);group B (the group of emphysema),the rats were exposed to cigarefte smoke two times every day, about 30 min each time; group C (the group of emphysema and SIH), the exposure condition of group B for 28 days,then still exposed to cigarette smoke and also exposed to the mixed gas of 12.5% oxygen for three hours during sleeping time every day (the exposed time was divided into four periods,45 rain each period). All the rats were exposed for 56 days continuously,and then killed. The lungs and whole blood were collected. Lungs were cut and stained with HE. Semiquantitative image analytic method was used to observe the histopathological features, which were quantitatively analyzed hy pathological score of lungs,mean linear intercept(MLI) and mean alveolus number(MAN). And then, AT Ⅲ, FIB,vWF, F Ⅷwere tested. The data were analyzed by SPSS 11.5 system. Group comparison was determined by one-factor analysis of variance. The comparison between two groups was measured by SNK-q test. Results Group B and C got the histopathological changes of emphysema. Pathological score of lungs and MLI were significantly different from groups( F= 520. 751, F= 1037. 787, both P<0.05 ), B[( 52.00±5.16 ) %,(91.30± 2.26) μm] and C [( 73.30±3.86 )%, (99.32±2.81 ) μm] were significantly higher than A [(16.20±2.53)% ,(46. 79±3. 18) μm, q =28. 312, q=45. 157, q =50. 666, q =59. 800,all P<0.05],Cwas significantly higher than B(q= 16. 845, q=9. 135,all P<(0.05);MAN was significantly different from groups(F= 313. 304, P<0.05), B[(103.55+4.45)/mm2] and C [(83.70±4.72)/mm2] were significantly lower than A[(151.59±8.64)/mm2, q=24.358, q=34.426, both P<0.05],C was significantly lower than B(q=10. 068, P<0.05). Levels of AT Ⅲ were significantly different from groups (F=25.386, P<0.05),AT Ⅲ of B[(107.39+8.05)%] and C [(99.04±3.99)%] were significantly lower than A [(116.20 4±2.53) %, q=5.173, q= 10. 076, both P<0.05], C was significantly lower than B (q=4. 903, P<0.05). Levels of FIB, vWF and F Ⅷ were significantly different from groups(F= 112. 969, F=223. 666, F=70.04,all P <0. 05),B[(1774.60+72.49) mg/L,(64.45+ 1.72)%,(227. 78±10. 38) %] and C [(1984.00±73.69) mg/L), (71.96±1.85 )%, (303.25±32.93 )%] were significantly higher than A[(1515. 50+62. 79) mg/L,(56. 02±1.47)%,(200.36±4.88) %, q=11. 734, q=21. 218, q=15. 810, q=29. 895, q=4. 307, q=16. 161 ,all P<0.05],C was significantly higher than B(q =9. 483, q=14.085, q=11. 854, all P<0.05). Conclusions This study established a rat model of pulmonary emphysema with SIH successfully, It was found that the histopathological change of pulmonary emphysema aggravated while the pulmonary emphysema rats were exposed to sleep hypoxemia. The rats of emphysema were in the status of hypercoagulation which was more severe than normal. While exposed to sleep hypoxemia, the status became further more severe.