From the Department of Surgery (J.Y., S.S., S.E.N.), UT Health San Antonio, San Antonio; Coagulation and Blood Research, US Army Institute of Surgical Research (W.M., X.W., D.D., D.Z., A.P.C., J.B., S.E.N.), Fort Sam Houston, Texas; Department of Medicine, Uniformed Services University of the Health Sciences (D.B.), Bethesda, Maryland; and Department of Molecular Medicine (Z.L.), Greehey Children's Cancer Research Institute, UT Health San Antonio, San Antonio, Texas.
J Trauma Acute Care Surg. 2021 Dec 1;91(6):1002-1009. doi: 10.1097/TA.0000000000003381.
Gut dysbiosis, an imbalance in the gut microbiome, occurs after trauma, which may be ameliorated with transfusion. We hypothesized that gut hypoperfusion following trauma causes dysbiosis and that whole blood (WB) resuscitation mitigates these effects.
Anesthetized rats underwent sham (S; laparotomy only, n = 6); multiple injuries (T; laparotomy, liver and skeletal muscle crush injuries, and femur fracture, n = 5); multiple injuries and 40% hemorrhage (H; n = 7); and multiple injuries, hemorrhage, and WB resuscitation (R; n = 7), which was given as 20% estimated blood volume from donor rats 1 hour posttrauma. Baseline cecal mesenteric tissue oxygen (O2) concentration was measured following laparotomy and at 1 hour and 2 hours posttrauma. Fecal samples were collected preinjury and at euthanasia (2 hours). 16S rRNA sequencing was performed on purified DNA, and diversity and phylogeny were analyzed with QIIME (Knight Lab, La Jolla, CA; Caporaso Lab, Flagstaff, AZ) using the Greengenes 16S rRNA database (operational taxonomic units; 97% similarity). α and β diversities were estimated using observed species metrics. Permutational analysis of variance was performed for overall significance.
In H rats, an average decline of 36% ± 3.6% was seen in the mesenteric O2 concentration at 1 hour without improvement by 2 hours postinjury, which was reversed following resuscitation at 2 hours postinjury (4.1% ± 3.1% difference from baseline). There was no change in tissue O2 concentration in the S or T rats. β Diversity differed among groups for all measured indices except Bray-Curtis, with the spatial median of the S and R rats more similar compared with S and H rats (p < 0.05). While there was no difference in α diversity found among the groups, indices were significantly correlated with mesenteric O2 concentration. Members of the family Enterobacteriaceae were significantly enriched in only 2 hours.
Mesenteric perfusion after trauma and hemorrhage is restored with WB resuscitation, which influences β diversity of the gut microbiome. Whole blood resuscitation may also mitigate the effects of hemorrhage on intestinal dysbiosis, thereby influencing outcomes.
创伤后肠道菌群失调(肠道微生物组失衡),可能通过输血得到改善。我们假设创伤后肠道低灌注导致菌群失调,全血(WB)复苏可以减轻这些影响。
麻醉大鼠接受假手术(S;仅剖腹,n = 6);多发伤(T;剖腹、肝和骨骼肌挤压伤、股骨骨折,n = 5);多发伤和 40%出血(H;n = 7);以及多发伤、出血和 WB 复苏(R;n = 7),在创伤后 1 小时从供体大鼠给予 20%估计血容量。在剖腹后、创伤后 1 小时和 2 小时测量回肠肠系膜组织氧(O2)浓度。在损伤前和安乐死时(2 小时)收集粪便样本。使用 QIIME(加利福尼亚州拉霍亚的 Knight 实验室;亚利桑那州弗拉格斯塔夫的 Caporaso 实验室),基于 Greengenes 16S rRNA 数据库(操作分类单元;97%相似性)对纯化 DNA 进行 16S rRNA 测序,并分析多样性和系统发育。使用观察物种指标估计 α 和 β 多样性。进行可置换方差分析以确定总体显著性。
在 H 组大鼠中,在 1 小时时观察到肠系膜 O2 浓度平均下降 36%±3.6%,而在 2 小时时未见改善,在 2 小时时复苏后逆转(与基线相比差异为 4.1%±3.1%)。S 或 T 组大鼠的组织 O2 浓度没有变化。除了 Bray-Curtis 外,所有测量指标的组间β 多样性均不同,S 和 R 大鼠的空间中位数与 S 和 H 大鼠更为相似(p < 0.05)。虽然组间α 多样性无差异,但指标与肠系膜 O2 浓度显著相关。肠杆菌科的成员仅在 2 小时时明显富集。
创伤和出血后的肠系膜灌注通过 WB 复苏得到恢复,这影响了肠道微生物组的β 多样性。全血复苏还可能减轻出血对肠道菌群失调的影响,从而影响结果。