Objective:To investigate the effects of Shenfu Injection(SFI)on endothelial damage in a porcine model of hemorrhagic shock(HS).Methods:After being bled to a mean arterial pressure of40±3 mm Hg and held for 60 min,32 pigs were treated with a venous injection of either shed blood(transfusion group),shed blood and saline(saline group),shed blood and SFI(SEI group)or without resuscitation(sham group).Venous blood samples were collected and analyzed at baseline and 0,1,2,4,and 6 h after HS.Tumor necrosis factor-α(TNF-α),serum interleuking(IL)-6,and IL-10 levels were measured by enzymelinked immunosorbent assay(ELISA);expressions of vascular cell adhesion molecule-1(VCAM-1),intercellular adhesion molecule 1(ICAM-1),von Willebrand factor(vWF),plasminogen activator inhibitor-1(PAI-1)and Bcl-2,Bax,and caspase-3 proteins were determined by Western blot.Results:The serum level of TNF-αin the SFI group was significantly lower than in the other groups at 0,1,and 2 h after HS,while the level of IL-6 was lower at 4 and 6 h compared with the saline group(P<0.01 or P<0.05).The concentration of serum IL-10 was significantly higher in the SFI group than in the other groups at 0,1,4,and 6 h after HS(P<0.01).Western blot and immunohistochemistry of vascular tissue showed that the expression of caspase-3 was downregulated,and that of Bcl-2 and Bax was upregulated in the SFI group compared to other groups(P<0.05).Conclusion:SFI attenuated endothelial injury in the porcine model of HS by inhibiting cell apoptosis,suppressing the formation of proinflammatory cytokines,and reducing endothelial activation.
目的通过猪心脏骤停窒息模型探讨参附注射液对此类肺损伤保护作用的可能机制。方法34只健康近交系五指山小型猪使用气管插管夹闭方法制作窒息型心脏骤停动物模型及行标准的心肺复苏术,其中18只成功恢复自主循环(return of spontaneous circulation,ROSC),使用随机数字表法分为两组:参附组(n=9):于ROSC后即刻以参附注射液0.24mg/min的剂量持续静脉泵入直至复苏后6h;盐水组(n=9):于ROSC后即刻持续静脉泵入相同剂量及速度的生理盐水直至复苏后6h。通过血气分析仪测量基础状态、ROSC即刻、ROSC后15min、30min、1h、2h、4h及6h的氧代谢及呼吸力学指标[包括:氧合指数(OI)、呼吸指数(RI)、氧输送(DO2)、氧消耗(VO2)、氧摄取(O2ER)、二氧化碳分压(PaCO2)及血乳酸(LAC),并监测同一时刻动物的肺顺应性(Cdyn)、气道阻力(Raw)、血管外肺水指数(EVLWI)和肺血管通透性指数(PVPI)];酶联免疫吸附法测定Na+-K+-ATP酶活性、Ca2+-ATP酶活性、SOD及MDA含量、TNF-α、IFN-γ和IL-4浓度,计算IFN-γ/IL-4值;利用TUNEL法检测细胞凋亡并计算凋亡指数(AI);免疫组织化学法检测Bcl-2、Bax蛋白浓度,计算BAX/BCL-2值;Western blot法检测Caspase-3蛋白定量。结果至ROSC后6h,参附组存活率为88.9%(8/9),盐水组存活率为66.7%(6/9);参附组平均生存时间(5.77±0.71)h长于盐水组(4.77±0.59)h,两组比较差异无统计学意义(P>0.05)。与基础状态比较,两组的OI和Cdyn在ROSC即刻明显降低(P<0.05),而RI、DO_2、VO_2、O_2ER、Raw、EVLWI、PVPI、PaCO_2和LAC在ROSC即刻则显著升高(P<0.05),各指标不论升高或降低均随时间延长而恢复;与盐水组比较,参附组在ROSC后的各个时间点,OI、Cdyn、DO_2、VO_2和O2ER显著高于盐水组(P<0.05,P<0.01),而RI、Raw、EVLWI、PVPI、PaCO_2和LAC则低于盐水组(P<0.05,P<0.01)。与盐水组比较,参附组Na^+-K^+-ATP酶、Ca^(2+)-ATP酶、SOD水平、IFN-γ水平、IFN-γ/IL-4值及Bcl-2浓度升高,而M
Objective： To investigate whether Shen-Fu Injection（参附注射液, SFI） reduces post-resuscitation immune dysfunction in a porcine model of cardiac arrest by modulating apoptosis of regulatory T lymphocytes（Treg） in the spleen. Methods： After 8-min untreated ventricular fibrillation and 2-min basic life support, 24 pigs were divided into 3 groups with a random number table, i.e. SFI group, epinephrine（EP） group, and saline（SA） group（8 in each group）, which received central venous injection of SFI（1.0 m L/kg）, EP（0.02 mg/kg） and SA, respectively. The same procedure without CA initiation was achieved in the sham-operated（sham） group（n=6）. After successful return of spontaneous circulation（ROSC）, apoptosis rate of splenic Treg was detected by flow cytometry; and the m RNA expression of forkhead/winged helix transcription factor（Foxp3） of splenic Treg was detected by real time-polymerase chain reaction; and the levels of interleukin-4（IL-4） and interferon-γ（IFN-γ） in porcine splenic Treg were detected by using enzyme-linked immunosorbent assay（ELISA）. Results： Compared with the sham group, the apoptosis rate of Treg was significantly decreased, and the levels of Foxp3 m RNA expression, IFN-γ, IL-4 and IFN-γ/IL-4 were increased in the SA group（P〈0.05 or P〈0.01）. Compared with the EP and SA groups, SFI treatment increased the apoptosis rate of Treg and reduced the levels of Foxp3 m RNA expression, IFN-γ and IFN-γ/IL-4（P〈0.05）. Conclusions： SFI has significant effects in attenuating post-resuscitation immune dysfunction by modulating apoptosis of Treg in the spleen.
Background:Cardiac arrest(CA)is a terminal event that results in a range of pathophysiological changes in the body,most notably,systemic ischemia-reperfusion injury.The hypothalamic-pituitary-adrenal(HPA)axis is an important neuroendocrine system that modulates adrenocortical hormone release.This study was designed to investigate the changes in HPA-related hormone levels after successful cardiopulmonary resuscitation(CPR)and to explore possible etiologies to provide a basis for relevant clinical research.Methods:We collected the clinical data of 96 patients with CA admitted to the Emergency Department of Beijing Chaoyang Hospital,Capital Medical University,between January 2016 and May 2017.Serum samples were collected 6,24,and 72 hours after restoring spontaneous circulation(ROSC).The data were compared with those of the healthy control group(n=50).An enzyme-linked immunosorbent assay(ELISA)was performed to measure copeptin,adrenocorticotropic hormone(ACTH),corticotropin-releasing hormone(CRH),and total cortisol.Demographic data were collected for both groups.For the CPR group,clinical data and the end-of-study cerebral performance category(CPC)were analyzed.Patients were followed up through day 28.Death or survival after day 28 was used as the study endpoint.Simple values were expressed as medians and quartiles or ratios(%)for statistical analysis.Continuous variables are expressed as mean±standard deviation.Categorical variables were expressed as frequencies and percentages.The mean values of normally distributed measurement data were analyzed using 1-way analysis of variance(ANOVA)for among-group comparisons and the least significant difference(LSD)test for between-group comparisons.SPSS v17(SPSS,Chicago,IL,USA)was used for statistical analysis,and P<0.05 was considered statistically significant.Results:No significant between-group differences were observed in terms of age or sex.The 28-day mortality rate in the CPR group was 71%.ACTH and CRH levels were significantly lower in the CPR group than in the health
Background： Acute kidney injury （AKI） frequently occurs in cardiopulmonary resuscitation patients. Studies comparing the effects of extracorporeal membrane oxygenation （ECMO） with conventional cardiopuhnonary resuscitation （CCPR） on AKI were rare. This study aimed to compare the effects of ECMO with those of CCPR on survival rate and AKI and explore the underlying mechanisms in a swine model of cardiac arrest （CA）. Methods： Sixteen male pigs were treated with ventricular fibrillation to establish CA model and then underwent CCPR （CCPR group, n = 8） or ECMO during cardiopulmonary resuscitation （ECPR group, n = 8）. The study endpoints were 6 h after return of spontaneous circulation （ROSC） or death. Serum and urine samples were collected at baseline and during the 6 h after ROSC. The biomarkers of AKI were detected by enzyme-linked immunosorbent assay. The apoptosis of renal tubular epithelial cells was discovered by transmission electron microscope （TEM） and terminal deoxynucleotidyl transferase dUTP nick end labeling assay. Apoptosis-related genes were detected by immune-staining and Western blotting. Data were compared by Student＇s t-test. Results： All pigs in ECPR group were successfully resuscitated with a higher 6-h survival rate （8/8） compared to CCPR group （6/8）. The expressions ofAKl biomarkers including neutrophil gelatinase-associated lipocalin （NGAL）, tissue inhibitor ofmetalloproteinase2 （TIMP2）, insulin-like growth factor-binding protein 7 （IGFBP7）, liver fatty acid-binding protein （LFABP）, and kidney injury molecule l （Kim-1） were all increased along with the time after ROSC in both groups and lower in ECPR group compared with CCPR group. Especially, products of urinary T1MP and IGFBP levels （TIMP＊IGFBP） were significantly lower at ROSC4 （0.58 ± 0.10 ng^2/ml^2 vs. 1.18 ± 0.38 ng^2/ml^2, t = 4.33, P =0.003） and ROSC6 （1.79 ±0.45 ng2^/ml^2 vs. 3.00 ±0.44 ng^2/ml^2, t = 5.49, P 〈 0.001）; urinary LFABP was significant
Objective：To investigate the action of Shen-Fu Injection（参附注射液,SFI） in regulating the expression of the serum complements and inflammatory cytokines synthesized and released in response to the stress of global ischemia accompanying cardiac arrest（CA） and resuscitation.Methods：Thirty pigs were randomly divided into the sham（n=6） and 3 returns of spontaneous circulation（ROSC） groups（n=24）.After 8-min untreated ventricular fibrillation and 2-min basic life support,24 pigs of the ROSC groups were randomized into three groups（n=8 per group）,which received central venous injection of SFI（SFI group）,epinephrine（EP group）,or saline（SA group）.Hemodynamic status and blood samples were obtained at 0,0.5,1,2,4,6,12,and 24 h after ROSC.Results：Serum concentrations of specific activation markers of the complement system C3,C4 and C5b-9 were increased during cardiopulmonary resuscitation th rough1 24 h after ROSC.There were intense changes of various pro-inflammatory cytokines and anti-inflammatory cytokines as early as 0.5 h after CA.Compared with the EP and SA groups,SFI treatment reduced the proinflammatory cytokines levels of interleukin（IL）-6,IL-8and tumor necrosis factor α（TNF-α,P〈0.05）,and increased the anti-inflammatory cytokine levels of IL-4 and IL-10（P〈0.05）.Further,SFI treatment decreased the values of C3,C4 and C5b-9 compared with the EP and SA groups.Conclusions：SFI,derived from the ancient Chinese medicine,has significant effects in attenuating post-resuscitation immune dysfunction by modulating the expression of complements and cytokines levels.The current study provided an experimental basis for the clinical application of a potential pharmacologic target for post resuscitation immune dysfunction.
