| 尹园缘,余 炼,刘 颖,黄静雯,邹巍莹,程 扬,宾东华.象皮生肌膏对肛瘘术后创面修复及PI3K/Akt/mTOR信号通路的影响[J].现代生物医学进展英文版,2024,(5):813-821. |
| 象皮生肌膏对肛瘘术后创面修复及PI3K/Akt/mTOR信号通路的影响 |
| Exploring the Mechanism of Xiangpi Shengji Ointment Promoting Wound Repair after Anal Fistula Based on PI3K/Akt Pathway |
| Received:October 04, 2023 Revised:October 27, 2023 |
| DOI：10.13241/j.cnki.pmb.2024.05.003 |
| 中文关键词: PI3K/ Akt/mTOR信号通路 象皮生肌膏 肛瘘 创面修复 |
| 英文关键词: PI3K/Aktt/mTOR pathway Xiangpi Shengji ointment Anal fistula Wound repair |
| 基金项目:国家自然科学基金项目（81603634）；湖南省临床医疗技术创新引导项目（2021SK51416；2020SK51404）；湖南省中医药管理局重点课题（C2022019）；长沙市自然科学基金项目（Kq2202458）；湖南中医药大学校级课题重点项目（2019XJJJ034）；湖南中医药大学中西医结合一流学科开放课题重点项目（2020ZXYJH05）；湖南中医药大学中西医结合一流学科开放课题一般项目（2020ZXYJH57）；2022年湖南中医药大学校级研究生创新课题（2022CX161；2022CX140） |
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| 中文摘要: |
| 摘要 目的：探究象皮生肌膏对肛瘘切除术后大鼠创面的治疗作用及其与PI3K/Akt/mTOR信号通路的相关性。方法：将SD雄性大鼠随机分为假手术组、模型组、象皮生肌膏组、湿润烧伤膏组，共4组，每组10只。4组均采用0号钢丝制造肛瘘模型，造模成功后，除假手术组外，均在麻醉下行肛瘘切除术，创面保持开放，使之形成"开放、渗血、渗液、有脓性分泌物"的感染性肛瘘术后创面，假手术组保留瘘管，不予换药，模型组予以生理盐水冲洗、络合碘消毒后不予换药，治疗组分别予以相应药物进行创面换药，共10天。10天后采用常规面积检测法比较模型组、象皮生肌膏组、湿润烧伤膏组大鼠肛瘘术后创面的创面愈合率及创面肉芽组织覆盖率；HE染色观察各组大鼠肛周组织病理情况；ELISA检测各组大鼠血清中bFGF、EGF、VEGF的表达水平，WB检测比较各组大鼠肛周肉芽组织中PI3K、Akt、mTOR、p70 S6K、p-PI3K（S473）、p-AKT（S473）、p-mTOR（Ser2448）、p-p70 S6K的蛋白表达水平。结果：与模型组比较，治疗第3、7、10天象皮生肌膏组和湿润烧伤膏组的创面愈合率及创面肉芽组织填充率显著升高（P＜0.01）。病理切片显示，假手术组炎性细胞浸润较少，其余各组均可见不同程度的炎性细胞浸润，且创面区可见不同程度炎性修复型肉芽组织增生、胶原纤维新生及血管扩张；其中模型组病理切片显示大量炎性细胞浸润，血管扩张明显，并有明显的血管出血；象皮生肌膏组病理切片显示炎性细胞较少，成纤维细胞成熟且分布整齐，真皮层内胶原纤维丰富且排列整齐，可见血管新生，无明显血管扩张及出血；湿润烧伤膏组病理切片显示少量炎性细胞浸润及血管扩张，真皮层内可见成纤维细胞生成。ELISA检测结果显示，与假手术组比较，模型组血清中bFGF、EGF、VEGF的含量显著降低（P＜0.01）；与模型组比较，象皮生肌膏组血清中bFGF、EGF、VEGF的含量显著升高（P＜0.01），湿润烧伤膏组大鼠血清中EGF、VEGF的含量显著升高（P＜0.01）。WB检测结果显示，与假手术组比较，模型组肛周组织中p-PI3K、p-Akt 、p-mTOR、p-p70 S6K蛋白表达显著降低（P<0.01）；与模型组比较，象皮生肌膏组肛周组织中p-PI3K、p-Akt 、p-mTOR、p-p70 S6K蛋白表达显著升高（P<0.05）。结论：象皮生肌膏能有效促进肛瘘术后创面修复，减轻肛瘘术后创面炎症反应，促进创面肉芽生长，其促愈机制可能与PI3K/Akt/mTOR信号通路相关蛋白表达有关，其通过上调PI3K/Akt/mTOR信号通路相关蛋白激活PI3K/Akt/mTOR信号通路，进而加快创面修复进程。 |
| 英文摘要: |
| ABSTRACT Objective: To explore the therapeutic effect of Xiangpi Shengji Ointment on the wounds of rats after anal fistula resection and its correlation with PI3K/Akt/mTOR pathway. Methods: SD male rats were randomly divided into four groups: sham operation group, model group, Xiangpi Shengji ointment group, and MEBO group, with 10 rats in each group. All four groups were used to create an anal fistula model using No. 0 steel wire. After successful modeling, except for the sham operation group, anal fistula resection was performed under anesthesia, and the wound was kept open to form an infectious anal fistula postoperative wound that was "open, oozing blood, oozing fluid, and purulent secretion." The sham operation group retained the fistula without changing the dressing. The model group was rinsed with physiological saline and disinfected with complex iodine without changing the dressing. The treatment group was given corresponding drugs to change the dressing for the wound, respectively, A total of 10 days. After 10 days, the wound healing rate and granulation tissue coverage rate of rats in the model group, the Xiangpi Shengji ointment group, and the MEBO group after anal fistula surgery were compared using conventional area detection methods; HE staining was used to observe the pathological changes of perianal tissues of rats in each group; The expression levels of bFGF, EGF, and VEGF in serum of rats in each group were detected by ELISA, and the protein expression levels of PI3K, Akt, mTOR, p70 S6K, p-PI3K (S473), p-AKT (S473), p-mTOR (Ser2448), and p-p70 S6K in perianal granulation tissue of rats in each group were compared by WB detection. Results: Compared with the model group, the wound healing rate and granulation tissue filling rate of the Xiangpi Shengji Ointment group and the MEBO group significantly increased on the 3rd, 7th, and 10th day of treatment (P<0.01); Pathological sections showed that in the sham operation group, there was less inflammatory cell infiltration, while in the other groups, there were different degrees of inflammatory cell infiltration, and different degrees of inflammatory repair type granulation tissue proliferation, collagen fiber regeneration, and vascular dilation were visible in the wound area. Pathological sections of the model group showed a large amount of inflammatory cell infiltration, significant vascular dilation, and significant vascular bleeding. Pathological sections of the Xiangpi Shengji Paste group showed fewer inflammatory cells, Fibroblasts are mature and evenly distributed, and collagen fibers are abundant and neatly arranged in the dermis. Angiogenesis is visible, without significant vasodilation and bleeding. Pathological sections in the MEBO group show a small amount of inflammatory cell infiltration and vasodilation, and fibroblasts are visible in the dermis. The results of ELISA showed that the levels of bFGF, EGF, and VEGF in the serum of the model group were significantly lower than those of the sham operation group (P<0.01); Compared with the model group, the content of bFGF, EGF, and VEGF in the serum of the Xiangpi Shengji Ointment group was significantly increased (P<0.01), while the content of EGF, VEGF in the serum of the rats in the MEBO group was significantly increased (P<0.01). The WB test results showed that compared with the sham operation group, the expression of p-PI3K, p-Akt, p-mTOR, and p-p70 S6K protein in the perianal tissue of the model group was significantly decreased (P<0.01); Compared with the model group, the expression of p-PI3K, p-Akt, p-mTOR, and p-p70 S6K protein in perianal tissue in the Xiangpi Shengji ointment group was significantly increased (P<0.05). Conclusion: Xiangpi Shengji Ointment can effectively promote wound repair after anal fistula surgery, alleviate inflammatory reactions on the wound after anal fistula surgery, and promote the growth of wound granulation. Its healing mechanism may be related to the expression of PI3K/Akt/mTOR signal pathway related proteins, which can accelerate the process of wound repair by upregulating PI3K/Akt/mTOR signal pathway related proteins, activating PI3K/Akt/mTOR signal pathway. |
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