Department of Surgery, Boston University School of Medicine, Boston, MA.
Department of Surgery, Boston University School of Medicine, Boston, MA.
Surgery. 2014 Feb;155(2):234-44. doi: 10.1016/j.surg.2013.08.018. Epub 2013 Nov 14.
We previously demonstrated that postoperative peritoneal injury and inflammation contribute to adhesiogenesis. Recent evidence suggests that in addition to their role of interfering with the acetylation status of nuclear histone proteins, histone deacetylase inhibitors (HDACIs) including valproic acid (VPA) can target nonhistone proteins to resolve inflammation and modulate immune cells. We hypothesized that HDACIs could reduce adhesions.
Seventy-two rats underwent laparotomy with creation of 6 peritoneal ischemic buttons to induce adhesions. A single intraperitoneal (IP) dose of 50 mg/kg VPA was administered intraoperatively, whereas controls received vehicle. To evaluate the timing, 25 rats underwent ischemic button creation with either an intraoperative or a delayed IP dose of VPA at 1, 3, or 6 hours postoperatively. On postoperative day 7, adhesions were quantified. To investigate mechanisms, ischemic buttons were created in 24 rats and either VPA or saline was administered in 1 intraoperative dose. At 3 or 24 hours later, peritoneal fluid was collected and fibrinolytic activity measured. Alternatively, button tissue was collected 30 minutes postoperatively to measure tissue factor, fibrinogen, and vascular endothelial growth factor (VEGF) by real-time polymerase chain reaction or Western blot.
A single intraoperative dose of VPA reduced adhesions by 50% relative to controls (P < .001). Delayed dosing did not reduce adhesions. In operated animals, peritoneal fibrinolytic activity was not different between groups. Tissue factor mRNA was downregulated by 50% (P = .02) and protein by 34% (P < .01) in animals administered VPA versus saline. VPA decreased fibrinogen protein by 56% and VEGF protein by 25% compared with saline (P = .03).
These findings suggest that VPA rapidly reduces the extravasation of key adhesiogenic substrates into the peritoneum. A single, intraoperative intervention provides an ideal dosing strategy and indicates an exciting new role for HDACIs in adhesion prevention.
我们之前的研究表明，术后腹膜损伤和炎症会导致粘连形成。最近的证据表明，除了干扰核组蛋白蛋白的乙酰化状态外，组蛋白去乙酰化酶抑制剂（HDACIs），包括丙戊酸（VPA），还可以靶向非组蛋白蛋白来缓解炎症并调节免疫细胞。我们假设 HDACIs 可以减少粘连。
72 只大鼠接受剖腹术并创建 6 个腹膜缺血按钮以诱导粘连。术中单次腹腔内（IP）给予 50mg/kg VPA 剂量，而对照组给予载体。为了评估时间，25 只大鼠在术后 1、3 或 6 小时进行术中或延迟 IP 给予 VPA 剂量以创建缺血按钮。术后第 7 天，量化粘连。为了研究机制，在 24 只大鼠中创建缺血按钮，并在术中给予 VPA 或生理盐水单次剂量。在 3 或 24 小时后，收集腹膜液并测量纤维蛋白溶解活性。或者，在术后 30 分钟收集按钮组织以通过实时聚合酶链反应或 Western blot 测量组织因子、纤维蛋白原和血管内皮生长因子（VEGF）。
单次术中给予 VPA 可使粘连相对于对照组减少 50%（P<0.001）。延迟给药不能减少粘连。在手术动物中，各组间腹膜纤维蛋白溶解活性无差异。与生理盐水组相比，给予 VPA 的动物组织因子 mRNA 下调 50%（P=0.02），蛋白下调 34%（P<0.01）。VPA 使纤维蛋白原蛋白减少 56%，VEGF 蛋白减少 25%，与生理盐水相比（P=0.03）。
这些发现表明，VPA 可迅速减少关键致粘连底物渗出到腹膜中。单次术中干预提供了理想的给药策略，并表明 HDACIs 在预防粘连方面具有令人兴奋的新作用。
