Migaly John, Lieberman Jonathan, Long Walter, Fisher Carol, Rolandelli Rolando H
Department of Surgery, Temple University School of Medicine, Philadelphia, Pennsylvania, USA.
Dis Colon Rectum. 2004 Oct;47(10):1699-705. doi: 10.1007/s10350-004-0647-3.
Transforming growth factor-beta1 plays a central role in colonic repair. We examined the temporal effect of vector-mediated transfer of transforming growth factor-beta1 on colonic anastomotic healing.
Male Sprague-Dawley rats (n = 24) underwent transection of the distal colon and single-layer anastomosis. Proximal to the anastomosis, the colon was again transected and a colostomy was matured proximally. The distal colon was intubated with a silicone catheter, tunneled along subcutaneous tissues, and connected to a swivel apparatus for postoperative luminal infusion. Rats were randomized into four groups (n = 6 each). Two control groups received 10(10) plaque-forming units of a Type 5 E1-deleted adenovirus carrying the bacterial beta-galactosidase gene either immediately following surgery or on postoperative Day 3. The treatment groups received transforming growth factor-beta1 with the same viral construct at parallel time points. On postoperative Day 6, anastomotic bursting pressure and site were determined in situ with the anastomotic tissue subsequently harvested and analyzed by enzyme-linked immunosorbent assay for beta-galactosidase and transforming growth factor-beta1.
When compared with its corresponding control, the group that received the transforming growth factor-beta1 gene on postoperative day 3 had a significantly higher bursting pressure (mmHg; 119 +/- 16 vs. 160 +/- 12, mean +/- SD; P = 0.001). While the majority of colons (5/6) from the control group burst at the anastomosis, none of the colons in the group that received transforming growth factor-beta1 on day 3 burst at the anastomotic site (P = 0.007). Beta-Galactosidase levels (pg/ml) in anastomotic tissue were significantly increased in both control groups when compared with their respective treatment groups (101 +/- 43 vs. 38 +/- 30, P = 0.01 when infused the day of surgery and 243 +/- 92 vs. 50 +/- 30, P = 0.009 when infused on day 3). Anastomotic levels of transforming growth factor-beta1 were also increased in the group receiving the transforming growth factor-beta1 gene on day 3 (214 +/- 66 vs. 135 +/- 24, P = 0.02).
Gene transfer into the healing colonic anastomosis can be effectively achieved via intraluminal administration of adenoviral vectors. Transfer of transforming growth factor-beta1 increased the strength of colonic anastomoses when given at Day 3 but not at Day 0, demonstrating its diverse effects in the wound healing sequence. Thus, gene transfer of transforming growth factor-beta1 may avoid the need for a diverting stoma in cases of rectal surgery and impaired healing resulting from chemotherapy or radiation.
转化生长因子β1在结肠修复中起核心作用。我们研究了载体介导的转化生长因子β1转移对结肠吻合口愈合的时间效应。
雄性Sprague-Dawley大鼠（n = 24）接受远端结肠横断和单层吻合术。在吻合口近端，再次横断结肠并在近端做成熟的结肠造口术。用硅胶导管插入远端结肠，沿皮下组织隧道化，并连接至旋转装置以便术后进行腔内输注。大鼠被随机分为四组（每组n = 6）。两个对照组在术后即刻或术后第3天接受携带细菌β-半乳糖苷酶基因的5型E1缺失腺病毒的10(10)个噬斑形成单位。治疗组在平行时间点接受携带相同病毒构建体的转化生长因子β1。术后第6天，原位测定吻合口破裂压力和部位，随后采集吻合口组织并通过酶联免疫吸附测定法分析β-半乳糖苷酶和转化生长因子β1。
与相应对照组相比，术后第3天接受转化生长因子β1基因的组具有显著更高的破裂压力（mmHg；119±16对160±12，平均值±标准差；P = 0.001）。虽然对照组的大多数结肠（5/6）在吻合口处破裂，但术后第3天接受转化生长因子β1的组中没有结肠在吻合口部位破裂（P = 0.007）。与各自的治疗组相比，两个对照组吻合口组织中的β-半乳糖苷酶水平（pg/ml）均显著升高（手术当天输注时为101±43对38±30，P = 0.01；术后第3天输注时为243±92对50±30，P = 0.009）。术后第3天接受转化生长因子β1基因的组中吻合口处的转化生长因子β1水平也升高（214±66对135±24，P = 0.02）。
通过腔内给予腺病毒载体可有效实现基因转移至愈合中的结肠吻合口。术后第3天给予转化生长因子β1可增加结肠吻合口的强度，但术后第0天给予则无此效果，这表明其在伤口愈合过程中的作用具有多样性。因此，转化生长因子β1的基因转移可能避免直肠手术以及化疗或放疗导致愈合受损情况下的转流造口需求。
