参附注射液对连续灌注联合控制性通气保存供肺保护作用的机制研究

Despite major a chievements in lung transplantation, we still face the persistent problem of a shortage of donor lungs. Current protocols for preserving donor lungs utilize hypothermic arrest and simple storage by a variety of preservation solutions. This is a suboptimal approach owing to the inevitable associated ischemia. Ischemic damage contributes to the risk of post-transplantation primary graft dysfunction and limits organ procurement and safe storage time. It has been suggested that donor lung preservation with continuous perfusion of oxygenated blood and ventilation provides better lung protection. Continuous perfusion and ventilation preservation, at least in theory, thereby overcomes the three major weaknesses in traditional static cold storage by ⑴ reducing ischemia/reperfusion injury, ⑵ reducing cold injury, and ⑶ allowing viability assessment. However, a perfusion device (mini-cardiopulmonary bypass) is necessary when preserves the donor lung in a ex vivo continuous perfusion state and prolonged cardiopulmonary bypass is associated with problems of blood cell damage, disorder of microcirculation, and inflammatory reaction. Disorder in hemodynamic and oxygen metabolism is occurs during lung perfusion and ventilation preservation, which is considered similar to gas collapse or blood collapse in early stage of shock by Chinese medicine. So we assume that adding a dose of Chinese medicinal, Shen-Fu injection with a function of replenishing qi to stop collapse, to perfusion solution should be benefit to the pulmonary protection. Shen-fu Injection for the treatment of syncope has a function of restoring yang to save from collapse. Our study and modern medicine have proved Shen-Fu injection can protection red blood cells, improve microcirculation, and relieve the inflammatory reaction. Shen-Fu injection has been clinically used for treatment of many kinds of diseases, especially being famous for its traditional cardiovascular protective effectiveness, such as shock, stabilizing blood pressure, and reducing acute lung injury. Therefore, we assume that adding Shen-Fu injection into the perfusate for preservation donor lung may ameliorate the harmful effects of cardiopulmonary bypass and provide a better pulmonary protection. The purpose of our study is to test the hypothesis that Shen-Fu injection could provide a better pulmonary protection by reducing blood cell damage, relieving disorder of microcirculation, and attenuating inflammatory reaction.

肺移植是肺疾病终末期主要治疗手段。供体的严重匮乏影响肺移植的数量和质量。实验证明连续氧合血灌注联合控制通气是目前供肺保存的最优方法，能扩大供肺标准，便于移植前组织配型和功能评估。但由于体外循环连续灌注保存期间发生了循环动力异常（与休克早期的气脱或血脱十分相似）。所以，保存过程中仍可以引起肺损伤。肺损伤的主要原因是红细胞破坏、微循环障碍以及炎症反应。我们研究发现在心脏不停跳保存期间参附注射液能保护心肌、保护红细胞、改善微循环和减轻炎症反应。但参附注射液的最佳血药浓度以及对微循环和红细胞的保护机理尚不明确。至今，尚未有参附注射液用于供肺连续氧合血灌注联合控制通气保存的报告。且相对于跳动的心脏，用肺脏来研究微循环更为方便。因此设想在肺连续氧合血灌注联合控制通气保存的灌注液中加入参附注射液能够提高供肺保存质量，且有助于进一步研究参附注射液对微循环和红细胞的保护机制。本课题拟以巴马小型猪为实验对象。

目的：肺移植是肺疾病终末期主要治疗手段。供体的严重匮乏是阻碍肺移植发展的重要因素,肺的保存质量是决定肺移植成败的关键。实验证明连续氧合血灌注联合控制通气是目前供肺保存的最优方法。但由于体外循环连续灌注保存期间发生了循环动力异常。所以，保存过程中仍可以引起肺损伤。肺损伤的主要原因是红细胞破坏、微循环障碍以及炎症反应。我们的研究发现在心脏不停跳保存期间参附注射液能保护红细胞、改善微循环和减轻炎症反应。因此设想在肺连续氧合血灌注联合控制通气保存的灌注液中加入有益气固脱作用的参附注射液应当有利于提高供肺保存质量。方法：选择体重25～30kg健康成年广西巴马小型猪18头，随机分成3组（n=6)。实验组1用含参附注射液（50ml，其中10ml用于预充，40ml微量泵持续注入）的去白细胞氧合血连续灌注联合控制通气保存供肺9h。9h后用全血灌注,3h末测定各种指标；实验组2用含参附注射液（100ml，其中20ml用于预充，80ml微量泵持续注入）的去白细胞氧合血连续灌注 联合控制通气保存供肺9h。9h后用全血灌注，3h末测定各种指标；对照组用去白细胞的氧合血连续灌注联合控制通气保存供肺9h。9h后用全血 灌注，3h末测定各种指标。结果：研究结果显示乳酸水平随着灌注时间延长，显著升高，说明长时间的灌注仍然存在肺组织灌注不足，组织酸性代谢产物堆积的问题。但是参附组和对照组间的乳酸水平比较在各个灌注时间点均没有统计学差异(P>0.05)。灌注液中的红细胞、白介素-6、C反应蛋白、血气分析指标（PH、氧分压、二氧化碳分压）组间各时点之间比较无统计学差异(P>0.05)。肺组织病理学研究结果显示参附1组和参附2组总体上看炎症反应较对照组轻（淋巴细胞浸润更少），提示参附注射液对供体肺组织有一定的保护作用。结论：研究结果显示参附注射液对供体肺组织有一定的保护作用。