基于TOLL受体及PI3K/AKT/mTOR信号通路探讨愈口宁对复发性阿弗他溃疡大鼠炎性因子的调节

doi:10.13701/j.cnki.kqyxyj.2023.01.013

摘要/Abstract

摘要： 目的:探讨愈口宁针对复发性阿弗他溃疡(RAU)大鼠黏膜创面促进愈合作用及TLR4/PI3K/Akt/mTOR及相关通路的影响。方法:将40只SPF级SD大鼠随机分为4组:空白对照组、模型对照组、左旋咪唑组以及愈口宁组,每组10只。除空白对照组外,采用化学灼烧法对各组大鼠右侧颊黏膜灼烧,从而造成RAU大鼠模型,连续给药7 d。比较造模给药后0、1、3、7 d大鼠口腔溃疡面积的大小及愈合情况,给药7 d后麻醉下脱颈处死大鼠,将溃疡组织行HE染色。观察病理变化,Western blot检查溃疡组织中TOLL样受体4(TLR4)、磷脂酰肌醇-3-激酶(PI3K)、磷酸化磷脂酰肌醇-3-激酶(p-PI3K)、蛋白激酶B(Akt)、核因子-κB(NF-κB)、哺乳动物雷帕霉素靶蛋白(mTOR)蛋白表达量,酶联免疫吸附试验(ELISA)法检测血清白细胞介素-6(IL-6) 、肿瘤坏死因子-α(TNF-α)与炎症因子水平。结果:与空白对照组比较,余下3组大鼠均出现口腔溃疡,治疗后愈口宁组及左旋咪唑组大鼠溃疡面愈合情况良好,而模型组可见溃疡瘢痕。模型组大鼠血清中IL-6、TNF-α浓度显著上升,其中TLR4、Akt、NF-κB、mTOR表达量上调,p-PI3K/PI3K显著下调(P<0.01)。愈口宁组及左旋咪唑组与模型对照组比较,IL-6、TNF-α的含量明显下降(P<0.01),TLR4、Akt、NF-κB、mTOR表达量显著下调(P<0.01),p-PI3K/PI3K显著上调(P<0.01)。结论:愈口宁可以帮助RAU大鼠口腔溃疡的愈合,并且通过抑制TLR4/PI3K/Akt/mTOR通路的活性,降低炎性因子水平,改善口腔黏膜的损伤及炎症。

关键词: 愈口宁, 复发性阿弗他溃疡, TOLL受体, PI3K/Akt通路, 炎症因子

Abstract: Objective: To investigate the healing-promoting effect of yukouning on mucosal wounds and its effect on TLR4/PI3K/Akt/mTOR and related pathways in rats with RAU. Methods: Forty SPF-grade SD rats were divided into four groups randomly: blank control group, model control group, levamisole group, and yukouning group, with 10 rats in each group. Excluding the blank control group, the remaining three groups of rats were established by chemical cautery method for RAU modeling and the drugs were administered continuously for 7 days. The size and healing of oral ulcer surface of the rats were compared at day 0, 1, 3, and 7 after modeling administration. The rats were executed after 7 days of administration. The histopathological changes of oral mucosa of rats were observed by hematoxylin-eosin (HE) staining. Western blot was used to detect the expression of TOLL-like receptor4 (TLR4), phosphatidylinositol-3-kinase (PI3K), phosphorylated phosphatidylinositol-3-kinase (p-PI3K), protein kinase B (Akt), nuclear factor-κB (NF-κB), and mammalian target of rapamycin (mTOR) protein in the ulcerated tissues. The levels of tumor necrosis factor (TNF-α) and serum interleukin-6 (IL-6) inflammatory factors were measured by enzyme-linked immunosorbent assay (ELISA). Results: Except the blank control group, the other three groups all developed oral ulcers. After treatment, the ulcer surface of yukouning group and levamisole group healed well, while the ulcer scar was seen in the model group. In serum of model group, IL-6 and TNF-α were significant increased in concentration. Among them, TLR4, Akt, NF-κB, and mTOR were upregulated, and p-PI3K/PI3K was remarkable down-regulated (P<0.01). The contents of IL-6 and TNF-α in the yukouning and levamisole groups were significantly decreased compared with that of model control group (P<0.01), in which the expressions of TLR4, Akt, NF-κB, and mTOR were significantly downregulated (P<0.01) and p-PI3K/PI3K were significantly upregulated (P<0.01). Conclusion: Yukouning can help heal of oral ulcers in rats and reduce the level of inflammatory factors and improve oral mucosal damage and inflammation by inhibiting the activity of TLR4/PI3K/Akt/mTOR pathway.

Key words: yukouning, recurrent aphthous ulcer, TOLL receptor, PI3K/Akt pathway, inflammatory factors

刘佳文, 王汉明, 杨湘晖, 范志龙, 贺晓明, 甘雨娟, 刘荷婧. 基于TOLL受体及PI3K/AKT/mTOR信号通路探讨愈口宁对复发性阿弗他溃疡大鼠炎性因子的调节[J]. 口腔医学研究, 2023, 39(1): 68-73.

LIU Jiawen, WANG Hanming, YANG Xianghui, FAN Zhilong, HE Xiaoming, GAN Yujuan, LIU Hejing. Regulation of Yukouling on Inflammatory Cytokines in Rats with RAU Based on Toll Receptor and PI3K/AKT/mTOR Signal Pathway[J]. Journal of Oral Science Research, 2023, 39(1): 68-73.

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