象皮生肌膏对肛周脓肿术后创面愈合及 NLRP3/Caspase-1 通路的影响

The effects of Xiangpi Shengji Gao on wound healing and NLRP3/Caspase-1 pathway after perianal abscess surgery

关键词:  象皮生肌膏  肛周脓肿  创面愈合  NOD样受体蛋白3  半胱氨酸天冬氨酸蛋白酶1

Key words:  Xiangpi Shengji Gao  perianal abscess  wound healing  NOD-like receptor thermal protein domain-containing protein 3  Caspase-1

摘要:

[目的] 探索象皮生肌膏对肛周脓肿术后创面愈合及NOD样受体蛋白3（NLRP3）/半胱氨酸天冬氨酸蛋白酶1（Caspase-1）通路的影响。[方法] 建立肛周脓肿大鼠术后模型，并将其分为模型组、象皮生肌膏组（创面涂抹象皮生肌膏）、象皮生肌膏+激活剂组（象皮生肌膏涂抹+1 mg/kg NLRP3激活剂），并将执行假手术的大鼠作为对照组，每组10只；测量各组大鼠干预后4、9、14 d的创面并计算愈合率；酶联免疫吸附（ELISA）法检测大鼠血清白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）、白细胞介素-18（IL-18）、血管内皮生长因子（VEGF）、血管生成素-2（Ang-2）水平；苏木精-伊红（HE）染色法观察大鼠创面组织病理学形态；末端标记法（TUNEL）染色法检测大鼠创面组织细胞凋亡；免疫组化法检测大鼠创面组织新生毛细血管数、NLRP3及Caspase-1蛋白表达；Western blot法检测大鼠创面组织NLRP3、Caspase-1蛋白表达。[结果] 与对照组比较，模型组大鼠创面组织出血、血管扩张情况较为明显，炎性细胞浸润情况较多，创面愈合率、细胞凋亡率、新生毛细血管数、血清VEGF、Ang-2水平下降，而血清IL-1β、IL-6、IL-18水平及创面组织NLRP3、Caspase-1蛋白表达增加（P<0.05）；与模型组比较，象皮生肌膏组大鼠创面组织病理形态改善明显，创面愈合率、细胞凋亡率、新生毛细血管数及VEGF、Ang-2水平升高，IL-1β、IL-6、IL-18水平及NLRP3、Caspase-1蛋白表达降低（P<0.05）；在象皮生肌膏+激活剂组中，象皮生肌膏对肛周脓肿大鼠术后上述指标的改善作用可被NLRP3激活剂逆转（P<0.05）。[结论] 象皮生肌膏可通过抑制NLRP3/Caspase-1通路促进肛周脓肿术后创面愈合。

Abstract:

[Objective] To explore the effects of Xiangpi Shengji Gao on wound healing and NOD-like receptor thermal protein domain-containing protein 3（NLRP3）/Caspase-1 pathway after perianal abscess surgery. [Methods] A postoperative rat model of perianal abscess was established and assigned into model group，Xiangpi Shengji Gao group（applying Xiangpi Shengji Gao to the wound），and Xiangpi Shengji Gao+activator group（applying Xiangpi Shengji Gao+1 mg/kg NLRP3 activator）. Rats undergoing sham surgery were used as the control group，each with 10 rats. The wounds of rats in each group were measured at 4，9，and 14 days after intervention，and the healing rate was calculated. Enzyme-linked immunosorbent assay（ELISA） method was applied to detect the levels of interleukin-1β（IL-1β），interleukin-6（IL-6），interleukin-18（IL-18），vascular endothelial growth factor（VEGF），and angiopoietin-2（Ang-2） in rat serum. Hematoxylin-eosin（HE） staining method was applied to observe the pathological morphology of rat wound tissue. TdT-mediated dUTP Nick-End Labeling（TUNEL） staining method was applied to measure apoptosis of rat wound tissue cells. Immunohistochemistry was used to measure the number of newly formed capillaries，and the NLRP3，and Caspase-1 protein expression in rat wound tissue. Western blot was used to measure the NLRP3 and Caspase-1 proteins in rat wound tissue. [Results] Compared with the control group，the model group showed more obvious bleeding and vascular dilation in the wound tissue，more inflammatory cell infiltration，decreased wound healing rate，apoptosis rate，number of newly formed capillaries，serum VEGF and Ang-2 levels，while increased serum IL-1β，IL-6，IL-18 levels，and NLRP3 and Caspase-1 protein expression in the wound tissue（P<0.05）. Compared with the model group，the Xiangpi Shengji Gao group showed prominent improvement in the pathological morphology of rat wound tissue，increased wound healing rate，apoptosis rate，number of newly formed capillaries，VEGF，and Ang-2，while decreased IL-1β，IL-6，IL-18 levels，and NLRP3 and Caspase-1 protein expression（P<0.05）. In the Xiangpi Shengji Gao+activator group，the improvement effect of Xiangpi Shengji Gao on the above indicators in rats with perianal abscess after surgery could be reversed by NLRP3 activator（P<0.05）. [Conclusion] Xiangpi Shengji Gao can promote wound healing after perianal abscess surgery by inhibiting the NLRP3/Caspase-1 pathway.