α-1 抗胰蛋白酶可改善肠道上皮细胞模型吻合口愈合。
Alpha-1-antitrypsin improves anastomotic healing in intestinal epithelial cells model.
机构信息
Department of Pediatric Surgery, Hannover Medical School, Hannover, Germany.
Department of Respiratory Medicine, Hannover Medical School, Hannover, Germany.
出版信息
Pediatr Surg Int. 2024 Sep 30;40(1):258. doi: 10.1007/s00383-024-05841-7.
PURPOSE
Intestinal anastomosis is a routine procedure in pediatric surgery, with leakage being a significant complication. Human alpha1-antitrypsin (AAT), whose physiological serum concentrations range from 0.9-2.0 mg/ml, is known to accelerate wound healing and stimulate the expression of cell proliferation-related genes. We hypothesized that AAT might enhance anastomotic healing.
METHODS
In a monolayer of non-tumorigenic HIEC-6 epithelial cells derived from fetal intestine a scratch was created. Standard medium without (control) or with AAT (0.5 and 1 mg/ml) was added. Cells were observed using a Life-Cell Imaging System. Cell proliferation was assessed, and the expression of proliferation-related genes was measured by qRT-PCR.
RESULTS
In the presence of AAT, the scratch closed significantly faster. Cells treated with 1 mg/ml AAT showed 53% repopulation after 8 h and 97% after 18 h, while control cells showed 24% and 60% repopulation, respectively (p < 0.02). The treatment with AAT induced HIEC-6-cell proliferation and significantly increased the mRNA-expression of CDKN1A, CDKN2A, ANGPTL4, WNT3 and COL3A1 genes. AAT did not change the mRNA-expression of CXCL8 but decreased levels of IL-8 as compared to controls.
CONCLUSION
At physiological concentrations AAT accelerates the confluence of intestinal cells and increases cell proliferation. The local administration of AAT may bear therapeutic potential to improve anastomotic healing.
目的
肠吻合术是小儿外科的常规手术，其并发症之一是吻合口漏。人α1-抗胰蛋白酶（AAT）的生理血清浓度范围为 0.9-2.0mg/ml，已知其可加速伤口愈合并刺激细胞增殖相关基因的表达。我们假设 AAT 可能增强吻合口愈合。
方法
在源自胎儿肠的非致瘤性 HIEC-6 上皮细胞单层上划痕。加入不含 AAT（对照）或含 AAT（0.5 和 1mg/ml）的标准培养基。使用 Life-Cell 成像系统观察细胞。评估细胞增殖，并通过 qRT-PCR 测量与增殖相关的基因表达。
结果
在 AAT 的存在下，划痕明显更快地闭合。用 1mg/ml AAT 处理的细胞在 8 小时后有 53%的再增殖，在 18 小时后有 97%的再增殖，而对照细胞分别有 24%和 60%的再增殖（p < 0.02）。AAT 处理诱导 HIEC-6 细胞增殖，并显著增加 CDKN1A、CDKN2A、ANGPTL4、WNT3 和 COL3A1 基因的 mRNA 表达。与对照组相比，AAT 没有改变 CXCL8 的 mRNA 表达，但降低了 IL-8 的水平。
结论
在生理浓度下，AAT 加速肠细胞的融合并增加细胞增殖。AAT 的局部给药可能具有改善吻合口愈合的治疗潜力。
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