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[参附注射液对窒息性心脏骤停猪模型复苏后肺损伤的保护作用]

[Protective Effect of Shenfu Injection on Postresuscitation Lung Injury in a Porcine Model of As- physia-induced Cardiac Arrest].

作者信息

Yang Jun, Li Chun-Sheng, Wu Cai-Jun, Hang Chen-Chen

出版信息

Zhongguo Zhong Xi Yi Jie He Za Zhi. 2016 Aug;36(8):967-974.

Abstract

Objective To observe the protective effect of Shenfu Injection ( SFI) on post-resusci- tation lung injury in a porcine model of asphyxia-induced cardiac arrest. Methods Thirty-four anaesthe- tized Wuzhi Mountain inbred miniature piglets of both sexes were subjected to asphyxia by intubation clip- ping, followed by standard cardiopulmonary resuscitation. Eighteen successfully resuscitated pigs [with recovery of return of spontaneous circulation ( ROSC) ] were divided into the SFI group and the normal saline (NS) group according to random digit table, 9 in each group. SFI at 0. 24 mg/min was intravenously pumped to piglets in the SFI group immediately from ROSC to 6 h after resuscitation, while NS at 0. 24 mg/min was intravenously pumped to piglets in the NS group immediately from ROSC to 6 h after resusci- tation. Oxygen metabolism, respiratory mechanics indices including oxygenation index (ΟI) , respiration index ( RI) , oxygen delivery ( DO₂), oxygen consumption ( VO₂), oxygen extraction ratio (Ο₂ ER), PaCO₂, lactic acid (LAC) were detected using blood gas analyzer at basic state, immediately after ROSC, 15 and 30 min, 1, 2, 4, and 6 h after ROSC. Dynamic lung compliance (Cdyn) , airway resistance (Raw), external vascular lung water index (EVLWI) , pulmonary vascular permeability index (PVPI) were monitored at each aforesaid time point. Activities of Na+-K +-ATPase and Ca² +-ATPase, contents of SOD and MDA, concentrations of TNF-α, IFN-γ, and IL-4 were determined using ELISA.IFN-γ/IL-4 ratio was calculated. Cell apoptosis was detected using TUNEL and apoptotic index (Al) calculated. Protein concentrations of Bcl-2 and Bax were detected using immunohistochemical assay, and Bax/Bcl-2 ratio calculated. Caspase-3 protein was quantitatively detected using Western blot. Results The survival rate was 88. 9% (8/9) in the SFI group and 66. 7% (6/9) in the NS group at 6 h after ROSC. The mean survival time was (5. 77 ±0. 71) h in the SFI group, longer than that in the NS group [ (4. 77 ±0. 59) h, P >0. 05]. Compared with the basic state, 01 and Cdyn obviously decreased immediately after ROSC (P <0. 05) ; RI, DO₂, VΟ₂, O₂ER, Raw, EVLWI, PVPI, PaCO₂, and LAC obviously increased immediately after ROSC (P<0. 05). All indices were recovered as time went by. Compared with the NS group, ΟI, Cdyn, DO₂, VΟ₂, and Ο₂ ER at each time points after ROSC were significantly higher in the SFI group than in the NS group (P <0. 05, P <0. 01); RI, Raw, EVLWI, PVPI, PaCO₂, and LAC were significantly lower in the SFI group than in the NS group (P <0. 05, P <0. 01 ). Compared with the NS group, activities of Na'-K '-AT- Pase and Ca² +-ATPase, contents of SOD, level of IFN-γ, IFN-γ/IL-4 ratio, concentrations of Bcl-2 in- creased more; MDA, TNF-α, IL-4 level, Al, Bax/Bcl-2 ratio, Caspase-3 protein level decreased more (P <0. 05, P <0. 01). Conclusion SFI could improve cell energy metabolism, enhance antioxidant ca- pacity of cells, reduce the release of inflammatory mediators, regulate the Thl/Th2 balance, and attenu- ate cell apoptosis of lung tissue, thereby protecting post-resuscitation lung injury.

摘要

目的 观察参附注射液(SFI)对窒息性心脏骤停猪模型复苏后肺损伤的保护作用。方法 34只麻醉的五指山近交系雌雄微型仔猪,通过气管插管夹闭造成窒息,随后进行标准心肺复苏。18只成功复苏[恢复自主循环(ROSC)]的猪,根据随机数字表分为SFI组和生理盐水(NS)组,每组9只。SFI组仔猪从ROSC即刻至复苏后6 h,以0.24 mg/min的速度静脉泵注SFI,NS组仔猪从ROSC即刻至复苏后6 h,以0.24 mg/min的速度静脉泵注NS。在基础状态、ROSC即刻、ROSC后15和30 min、1、2、4和6 h,使用血气分析仪检测氧代谢、呼吸力学指标,包括氧合指数(OI)、呼吸指数(RI)、氧输送(DO₂)、氧消耗(VO₂)、氧摄取率(O₂ER)、PaCO₂、乳酸(LAC)。在上述各时间点监测动态肺顺应性(Cdyn)、气道阻力(Raw)、血管外肺水指数(EVLWI)、肺血管通透性指数(PVPI)。采用ELISA法测定Na⁺-K⁺-ATP酶和Ca²⁺-ATP酶活性、SOD和MDA含量、TNF-α、IFN-γ和IL-4浓度。计算IFN-γ/IL-4比值。采用TUNEL法检测细胞凋亡并计算凋亡指数(AI)。采用免疫组织化学法检测Bcl-2和Bax蛋白浓度,并计算Bax/Bcl-2比值。采用Western blot法定量检测Caspase-3蛋白。结果 ROSC后6 h,SFI组生存率为88.9%(8/9),NS组为66.7%(6/9)。SFI组平均生存时间为(5.77±0.71)h,长于NS组[(4.77±0.59)h,P>0.05]。与基础状态相比,ROSC即刻OI和Cdyn明显降低(P<0.05);RI、DO₂、VO₂、O₂ER、Raw、EVLWI、PVPI、PaCO₂和LAC在ROSC即刻明显升高(P<0.05)。随着时间推移,所有指标均恢复。与NS组相比,ROSC后各时间点SFI组的OI、Cdyn、DO₂、VO₂和O₂ER均显著高于NS组(P<0.05,P<0.01);RI、Raw、EVLWI、PVPI、PaCO₂和LAC在SFI组显著低于NS组(P<0.05,P<0.01)。与NS组相比,SFI组Na⁺-K⁺-ATP酶和Ca²⁺-ATP酶活性、SOD含量、IFN-γ水平、IFN-γ/IL-4比值、Bcl-2浓度升高更明显;MDA、TNF-α、IL-4水平、AI、Bax/Bcl-2比值、Caspase-3蛋白水平降低更明显(P<0.05,P<0.01)。结论 SFI可改善细胞能量代谢,增强细胞抗氧化能力,减少炎症介质释放,调节Th1/Th2平衡,减轻肺组织细胞凋亡,从而保护复苏后肺损伤。

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