硫化氢对组织因子诱导的家兔弥散性血管内凝血的影响

Effect of hydrogen sulfide on tissue factor-induced disseminated intravascular coagulation in rabbits

目的 探讨外源性硫化氢（H：S）对组织因子诱导的弥散性血管内凝血（DIC）家兔的保护作用及其机制。方法 将32只健康家兔按照随机数字表法分为正常对照组、硫氢化钠（NaHS）对照组、DIC模型组、Naris预处理组，每组8只。制模前10min，NaHS对照组和NaHS预处理组经耳缘静脉注射NaHS溶液3．4mg／kg（加生理盐水至5mL混匀溶解），正常对照组和DIC模型组注射等量生理盐水；10rain注射完毕后，DIC模型组及NaHS预处理组经耳缘静脉注射冻干兔脑组织因子悬液2mL／kg（生理盐水稀释至30mL，以1mL／min×5min、2mL／min×5rain、3mL／min×5rain的速度注人），正常对照组和NaHS对照组注射等量生理盐水。各组分别于注射组织因子前10rain及注射后3、5、8、10、13、15、45、85、135min颈总动脉采血3mL，检测凝血酶原时间（PT）、活化部分凝血活酶时间（APTT）、凝血酶时间（TT）、纤维蛋白原（FIB）、纤维蛋白原降解产物（FDP）及血小板计数（PLT）；同时观察肠系膜微循环。结果与正常对照组比较，DIC模型组注射组织因子5min时PT、APTT缩短，其变化率明显增大[PT：-8．3（-11．7^-5．3）％比1．3（-2．5．3．8）％，P〈0．01：APTT：-19．1（-30．4～-9．4）％比-2．6（-6．2～3．0）％，P〈0．05]；15min后PT、APTT延长，其变化率明显增大[PT：31．0（25．0～36．9）％比-1．3（-6．3—5．0）％，APTT：61．3（50．0～72．9）％比0．0（-10．0～10．0）％，均P〈0．01]；注射组织因子后TT逐渐缩短，FIB、PLT逐渐降低，在15min时其变化率明显增大[TT：-9．5（-12．0～-6．2）％比-2．0（-4．0—0．7）％，FIB：-4．3（-9．9～-2．2）％比-1．0（-5．8—4．3）％，PLT：-90．0（-93．4～-86．5）％比-1．0（-3．9～2．6）％，均P〈0．01]；注射组织因子后FDP测试板圆圈内均有乳胶颗粒凝集，且在3～15min乳胶颗粒凝集逐渐增强，之后逐渐减弱；肠系膜毛细血管血流速于10min内明显变慢，之后逐渐变快并伴有明显出血。NaHS预处理组注射组织因子5min时PT、APTT缩短，其变化率较DIC模型组明显减小[PT：-6．3（-8．6—0．0）％比-8．3（-11．7^-5．3）％，APTT：-13．6（-24．2～-2．3）％比-19．1（-30．4～-9．4）％，均P〈0．05]；15min后PT、APTr延长，但其变化率较DIC模型组明显减小[PT：10．1（3．8～15．2）％比31．0（25．0～36．9）％，P〈0．01；APTT：27．8（-15．8^-39．7）％比61．3（50．0～72．9）％，P〈0．05]；15min时TT缩短，FIB、PLT降低，但其变化率较DIC模型组明显减小[TT：-4．5（-7．8^-1．3）％比-9．5（-12．0^-6．2）％，P〈0．01：FIB：-3．3（-8．0～1．9）％比-4．3（-9．9～-2．2）％，P〈0．05；PLT：-58．8（-53．0～64．0）％比-90．0（-93．4～-86．5）％，P〈0．01]；各时间点乳胶颗粒凝集较DIC模型组明显减弱；肠系膜毛细血管血流速10min内逐渐变慢，但较DIC模型组快，之后变快，且出血明显减少。结论 H2S对实验性DIC有一定的防治作用，其作用机制可能与其抑制组织因子引起的凝血系统激活及血小板聚集有关。

Objective To investigate the protective effect of hydrogen sulfide （H2S） on tissue factor-induced disseminated intravascular coagulation （DIC） in rabbits and its mechanism. Methods Thirty-two healthy rabbits were randomly divided into four groups： normal control group, NariS control group, DIC model group, NariS pretreatment group （each, n = 8）. Ten minutes before model reproduction, rabbits in NariS control and pretreatment groups were given 3.4 mg/kg NariS （dissolved in normal saline to 5 mL） via ear vein, while rabbits in normal control and DIC model groups were given an equivalent volume of normal saline. Ten minutes later, rabbits in DIC model and Naris pretreatment groups were intravenously given tissue factor （TF） 2 mL/kg （dissolved in normal saline to 30 mL, at the speed of 1 mL/min for 5 minutes, 2 mL/min for 5 minutes, and 3 mL/min for 5 minutes）, and rabbits in normal control and NariS control groups were given normal saline. 3 mL of blood was collected 10 minutes before TF injection, and 3, 5, 8, 10, 13, 15, 45, 85, 135 minutes after TF injection for determination of prothrombin time （PT）, activated partial thromboplastin time （APTT）, thrombin time （TT）, fibrinogen content （FIB）, fibrin degradation products （FDP）, and platelet count （ PLT ）. Microcirculation in the mesentery was also observed under microscope. Results Compared with normal control group, PT and APTT became shorter at 5 minutes after TF injection, and the rate of their change was increased [ PT： -8.3 （-11.7 to -5.3）% vs. 1.3 （-2.5 to 3.8）%, P 〈 0.01; APTT： -19.1 （-30.4 to -9.4）% vs. -2.6 （-6.2 to 3.0）%, P 〈 0.05]. PT and APTF were prolonged 15 minutes after TF injection, and their changes were more significant [ PT： 31.0 （25.0 to 36.9）% vs. -1.3 （-6.3 to 5.0）%, APTY： 61.3 （50.0 to 72.9）% vs. 0.0 （-10.0 to 10.0）%, both P 〈 0.01 ] in DIC model group. TY was gradually reduced after TF injection, FIB and PLT were gradually decreased, and their changes were more obvious at 15 minutes in DIC model group compared with those in normal control group [TT： -9.5 （-12.0 to -6.2 ）% vs. -2.0 （-4.0 to 0.7 ）%, FIB： -4.3 （-9.9 to -2.2）% vs. -1.0 （-5.8 to 4.3 ）%, PLT： -90.0 （-93.4 to -86.5 ）% vs. -1.0 （-3.9 to 2.6）, all P 〈 0.01 ]. After TF injection, it appeared latex-like particles in FDP test board, and it was gradually increased within 3-15 minutes, and then it gradually became less marked. The rate of blood flow in mesenteric capillaries was decreased obviously within 10 minutes, and it became faster accompanying with obvious hemorrhage. PT and APTr in NariS pretreatment group became shortened 5 minutes after TF injection, and their rate of change was significantly decreased compared with that of DIC model group [PT： -6.3 （-8.6 to 0.0）% vs. -8.3 （-11.7 to -5.3 ）%, APTT： -13.6 （-24.2 to -2.3 ）% vs. -19.1 （-30.4 to -9.4）%, both P 〈 0.05 ], and prolonged at 15 minutes, and their rate of change was significantly decreased compared with that of DIC model group [ PT： 10.1 （3.8 to 15.2）% vs. 31.0 （25.0 to 36.9）%, P 〈 0.01; APTT： 27.8 （-15.8 to 39.7）% vs. 61.3 （50.0 to 72.9）%, P 〈 0.05]. TY, FIB, and PLT were reduced at 15 minutes in NariS pretreatment group, and their rate of change was markedly decreased compared with that of DIC model group [TI＇： -4.5 （-7.8 to -1.3）% vs. -9.5 （-12.0 to -6.2）%, P 〈 0.01; FIB： -3.3 （-8.0 to 1.9）% vs. -4.3 （ -9.9 to -2.2 ）%, P 〈 0.05; PLT： -58.8 （-53.0 to 64,0）% vs. -90.0 （-93.4 to -86.5）%, P 〈 0.01 ]. The rate of agglutination of latex particles in Naris pretreatment group was decreased significantly at each time point compared with DIC model group; mesenteric capillary blood flow slowed down gradually within 10 minutes, but it was faster as compared with the DIC model group. It became faster later, but bleeding was obviously less. Conclusion These results show that H2S protects against TF-induced DIC by inhibiting the activity of coagulation system and platelet aggregation.