| 摘要: |
| 目的：探讨象皮生肌膏在糖尿病肛瘘术后创面愈合中的作用及其作用机制与磷脂酰肌醇 3-激酶/蛋白激酶 B/哺乳动物雷帕霉素靶蛋白（phosphoinositide 3-kinase/protein kinase B/mechanistic target of rapamycin，PI3K / AKT / mTOR）信号通路之间的关联性。方法：将30只健康成年SD雄性大鼠随机选取6只设为正常肛瘘组，其余24只分为糖尿病假手术组（行糖尿病、肛瘘造模，不行瘘管切除术）及糖尿病肛瘘组、糖尿病象皮生肌膏组及糖尿病湿润烧伤膏组（造模同上，30 d后行瘘管切除术），每组6只。术后24 h开始换药，每天1次，连续10 d。换药第5、7、10天评估和记录创面面积与愈合率。末次给药次日麻醉大鼠，采集肛周瘘管组织、创面组织，苏木精-伊红染色（Hematoxylin and Eosin Staining，HE）分析组织病理学特征，实时荧光定量法（quantitative Reverse Transcription Polymerase Chain Reaction，qRT-PCR）评估碱性成纤维细胞生长因子（fibroblast growth factor-basic，bFGF）的mRNA转录水平，免疫组织化学检测技术（Immunohistochemistry，IHC）定位α-平滑肌肌动蛋白（α-smooth muscle actin，α-SMA）的表达，蛋白质免疫印迹检测（Western Blotting，WB）定量分析PI3K/AKT/mTOR信号通路关键蛋白的表达情况。结果：1）血糖及一般情况。糖尿病大鼠随机血糖为16.7～33.3 mmol/L；正常大鼠体型均衡，毛发光泽，瘘管处脓性分泌物少；糖尿病大鼠体型消瘦，毛发枯黄，瘘管处脓性分泌物较多。2）创面愈合情况及愈合率。正常肛瘘组创面面积第5、7天显著缩小，肉芽组织大量新生，至第10天基本愈合；糖尿病肛瘘组、糖尿病象皮生肌膏组、糖尿病湿润烧伤膏组创面愈合进程较正常肛瘘组慢，愈合率降低（P<0.05），创面肉芽组织较红；糖尿病肛瘘组创面缩小速度及愈合率显著低于糖尿病象皮生肌膏组、糖尿病湿润烧伤膏组（P<0.05）。3）组织病理学。糖尿病假手术组有少量炎性细胞浸润；正常肛瘘组炎性细胞浸润少，成纤维细胞致密，血管丰富，组织完整；糖尿病肛瘘组较正常肛瘘组炎症细胞浸润增多，血管扩张、炎症反应更明显，肉芽组织较少；糖尿病象皮生肌膏组、糖尿病湿润烧伤膏组上皮细胞层增厚，胶原纤维排列致密，毛细血管新生，创面修复较完整。4）指标检测结果。与糖尿病假手术组相比，糖尿病肛瘘组bFGF、α-SMA、p-PI3K/PI3K及p-AKT/AKT水平显著降低（P<0.05）；与正常肛瘘组相比，糖尿病肛瘘组bFGF、α-SMA、p-PI3K/PI3K及p-AKT/AKT水平显著降低（P<0.05）；与糖尿病肛瘘组相比，糖尿病象皮生肌膏组与糖尿病湿润烧伤膏组bFGF、α-SMA、p-PI3K/PI3K及p-AKT/AKT水平显著增高（P<0.05）；各组p-mTOR/mTOR差异无统计学意义（P>0.05）。结论：象皮生肌膏在糖尿病肛瘘术后创面修复方面具有确切疗效，其作用机制可能与PI3K/AKT/mTOR信号通路的激活有关。 |
| 关键词: 糖尿病 肛瘘术后 创面修复 象皮生肌膏 PI3K/AKT/mTOR |
| DOI： |
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| Effect of Xiangpi Shengji ointment on wound repair and the phosphoinositide-3 kinase/protein kinase-B/mammalian target of rapamycin signaling pathway in diabetic rats after surgery for anal fistula |
| OUYANG Limin,BIN Donghua |
| (The First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha 410007, Hunan, China) |
| Abstract: |
| Objective: To investigate the effect and mechanism of action of Xiangpi Shengji ointment on wound repair after surgery for anal fistula in the presence of diabetes, as well as its association with the phosphoinositide-3 kinase （PI3K）/protein kinase-B （AKT）/mammalian target of rapamycin （mTOR） signaling pathway. Methods: A total of 30 healthy adult male Sprague-Dawley rats were selected, among which 6 were randomly selected as normal anal fistula group, and the remaining 24 rats were used to establish a model of diabetes and anal fistula and were then divided into diabetes+sham-operation group （without fistula resection）, diabetes+anal fistula group, diabetes+Xiangpi Shengji ointment group, and diabetes+moisture-exposed burn ointment group, with 6 rats in each group, while fistula resection was performed for the latter three groups after 30 days. Dressing change was performed since 24 hours after surgery, once a day for 10 consecutive days, and wound area and healing rate were assessed and recorded on days 5, 7, and 10 of dressing change. The rats were anesthetized on the next day after the last administration to collect perianal fistula tissue and wound tissue, and HE staining was used to observe histopathological features; qRT-PCR was used to measure the mRNA transcription level of basic fibroblast growth factor （bFGF）; immunohistochemistry was used to localize the expression of alpha-smooth muscle actin （α-SMA）; Western blotting was used to quantitatively analyze the expression levels of the proteins in the PI3K/AKT/mTOR signaling pathway. Results: As for blood glucose and general status, the diabetic rats had a random blood glucose level of 16.7-33.3 mmol/L; the normal rats had a well-proportioned body, glossy fur, and a small amount of purulent discharge from the fistula, while the diabetic rats were emaciated with lusterless hair and a large amount of purulent discharge from the fistula. As for wound healing and healing rate, the normal anal fistula group had a significant reduction in wound area on days 5 and 7, with a large amount of new granulation tissues, and the wound was basically healed on day 10; compared with the normal anal fistula group, the diabetes+anal fistula group, the diabetes+Xiangpi Shengji ointment group, and the diabetes+moisture-exposed burn ointment group had slower wound healing, a significantly lower healing rate, and red wound granulation tissues; compared with the diabetes+Xiangpi Shengji ointment group and diabetes+moisture-exposed burn ointment group, the diabetes+anal fistula group had significantly lower wound reduction rate and healing rate （P < 0.05）. As for histopathology, the diabetes+sham-operation group had a small amount of inflammatory cell infiltration; the normal anal fistula group had little inflammatory cell infiltration with dense fibroblasts, abundant blood vessels, and intact tissue; compared with the normal anal fistula group, the diabetes+anal fistula group had an increase in inflammatory cell infiltration with vasodilation, significant inflammatory response, and a lower number of granulation tissues; the diabetes+Xiangpi Shengji ointment group and diabetes+moisture-exposed burn ointment group had the thickening of epithelial layer, dense arrangement of collagen fibers, capillary regeneration, and relatively complete wound repair. As for the measurement of related indicators, compared with the diabetes+sham-operation group, the diabetes+anal fistula group had significant reductions in the levels of bFGF, α-SMA, p-PI3K/PI3K, and p-AKT/AKT （P < 0.05）; compared with the normal anal fistula group, the diabetes+anal fistula group had significant reductions in the levels of bFGF, α-SMA, p-PI3K/PI3K, and p-AKT/AKT （P < 0.05）; compared with the diabetes+anal fistula group, the diabetes+Xiangpi Shengji ointment group and diabetes+moisture-exposed burn ointment group had significant increases in the levels of bFGF, α-SMA, p-PI3K/PI3K, and p-AKT/AKT （P < 0.05）; there was no significant difference in p-mTOR/mTOR between groups （P > 0.05）. Conclusion: Xiangpi Shengji ointment has a marked clinical effect on wound repair after surgery for anal fistula in the presence of diabetes, possibly by activating the PI3K/AKT/mTOR signaling pathway. |
| Key words: diabetes after surgery for anal fistula wound repair Xiangpi Shengji ointment phosphoinositide-3 kinase/protein kinase-B/mammalian target of rapamycin |
