Raschke P, Becker B F, Leipert B, Schwartz L M, Zahler S, Gerlach E
Physiologisches Institut, Universität München, Germany.
Basic Res Cardiol. 1993 Jul-Aug;88(4):321-39. doi: 10.1007/BF00800639.
The role of polymorphonuclear neutrophils (PMN) in the injury of the heart following ischemia and reperfusion is still controversial. The aim of this study was to investigate whether small numbers of PMN may cause myocardial dysfunction in an isolated system, how the resulting loss of function can be characterized and whether the formation of hypochlorous acid (HOCl) can be responsible for the PMN-mediated effect. Isolated working guinea pig hearts were subjected to a 90% reduction of coronary flow for 30 min, with or without intracoronary infusion of homologous PMN (approximately 1-2 x 10(5) cells/min, i.e. about 5-10% of normal blood count). This ischemia was followed by a 15 min reflow period in a non-working ("Langendorff") mode before work was resumed. In hearts perfused only with buffer, post-hypoxic heart function recovered to 75-80% of the initial value. Inclusion of unstimulated PMN did not further attenuate cardiac function. However, cardiac output was decreased to 42% of the initial value, provided thrombin (0.3 U/ml) and H2O2 (10(-5) M) were also present, and the retained PMN (about 10% of those infused) were additionally stimulated during reflow by application of FMLP (10(-6) M for 1 min). In these instances, coronary flow at any time of the experiment and release of lactate or purines during ischemia and reflow did not differ significantly between hearts perfused with or without PMN. There was no substantial release of myoglobin in controls and in PMN-treated hearts. Inotropic stimulation of the hearts with noradrenaline or exogenous Ca2+ caused a sustained increase in contractile force. However, the response was significantly reduced in PMN-perfused hearts in comparison to control hearts. The myocardial contents of high-energy phosphates with and without inotropic stimulation proved to be identical irrespective of whether experiments had been performed in the absence or presence of PMN. A similar loss of myocardial function as mediated by PMN could be produced by infusing chemically generated hypochlorous acid (HOCl, 5 x 10(-7) M for 10 min). Strikingly, that portion of the infused HOCl which actually reacted with cardiac tissue was comparable to the amount shown to be generated by stimulating 10(6) PMN retained in the coronary system (about 7 nmoles). Supplementing the perfusate with the scavengers L-methionine (10(-4) M) or uric acid (5 x 10(-4) M) prevented the attenuation of heart function provoked by PMN. The results indicate that small numbers of PMN, sufficiently activated, can depress cardiac function after 30 min of ischemia.(ABSTRACT TRUNCATED AT 400 WORDS)
多形核中性粒细胞(PMN)在缺血再灌注后心脏损伤中的作用仍存在争议。本研究的目的是调查少量PMN在离体系统中是否会导致心肌功能障碍,如何表征由此产生的功能丧失,以及次氯酸(HOCl)的形成是否可导致PMN介导的效应。将离体工作的豚鼠心脏冠状动脉血流减少90%,持续30分钟,同时进行或不进行冠状动脉内输注同源PMN(约1 - 2×10⁵个细胞/分钟,即正常血细胞计数的约5 - 10%)。在恢复工作前,这种缺血状态之后是15分钟的非工作(“Langendorff”)模式的再灌注期。在仅用缓冲液灌注的心脏中,缺氧后心脏功能恢复到初始值的75 - 80%。加入未刺激的PMN并没有进一步削弱心脏功能。然而,如果同时存在凝血酶(0.3 U/ml)和过氧化氢(10⁻⁵ M),并且在再灌注期间通过应用FMLP(10⁻⁶ M,持续1分钟)额外刺激保留的PMN(约为输注量的10%),则心输出量会降至初始值的42%。在这些情况下,在实验的任何时间点,灌注有或没有PMN的心脏之间的冠状动脉血流以及缺血和再灌注期间乳酸或嘌呤的释放没有显著差异。在对照组和PMN处理的心脏中均没有大量肌红蛋白释放。用去甲肾上腺素或外源性Ca²⁺对心脏进行变力刺激会导致收缩力持续增加。然而,与对照心脏相比,PMN灌注的心脏中的反应明显降低。无论实验是在没有PMN还是有PMN的情况下进行,有或没有变力刺激时心肌高能磷酸盐的含量都被证明是相同的。通过输注化学合成的次氯酸(HOCl,5×10⁻⁷ M,持续10分钟)可产生与PMN介导的类似的心肌功能丧失。令人惊讶的是,实际与心脏组织反应的输注HOCl的量与刺激冠状动脉系统中保留的10⁶个PMN所产生的量相当(约7纳摩尔)。用清除剂L - 蛋氨酸(10⁻⁴ M)或尿酸(5×10⁻⁴ M)补充灌注液可防止PMN引起的心脏功能减弱。结果表明,少量充分活化的PMN在缺血30分钟后可降低心脏功能。(摘要截短至400字)