加味丹玄口康对大鼠口腔黏膜下纤维化及其自噬水平的影响

doi:10.20047/j.issn1673-7210.2023.24.02

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摘要目的探讨加味丹玄口康对大鼠口腔黏膜下纤维化（OSF）及其自噬水平的影响。方法选取SPF级SD雄性大鼠24只（180～200 g，4～8周龄），采用随机数字表法将其分为空白组、模型组和加味丹玄口康组，每组8只。模型组和加味丹玄口康组在大鼠双侧颊黏膜组织注射槟榔碱0.2 ml（10 mg/ml），1次/2 d，同时每天使用小牙刷沾取适量槟榔碱均匀施力刷大鼠双侧颊黏膜，持续8周，观察到大鼠颊黏膜出现白色病变确认造模成功，空白组不做干预。造模成功后，加味丹玄口康组以8 ml/（kg·d）加味丹玄口康灌胃，空白组和模型组灌胃等量0.9%氯化钠溶液，三组均灌胃4周。实验后取三组双侧颊黏膜进行苏木精-伊红染色和Masson染色观察病理变化和胶原纤维沉积情况，透射电镜观察三组自噬小体数量。采用免疫组织化学染色检测三组上皮钙黏素和微管相关蛋白轻链3（LC3）阳性细胞表达情况；Western blot检测上皮钙黏素、LC3-Ⅱ和选择性自噬接头蛋白p62的表达；RT-PCR检测LC3-Ⅱ和p62 mRNA表达。结果模型组口腔黏膜上皮明显萎缩，胶原纤维显著变粗、增多，上皮层变薄，黏膜下层胶原纤维大量沉积，加味丹玄口康组口腔黏膜上皮层较厚，胶原纤维减少，上皮层萎缩程度减轻。模型组自噬小体数量低于空白组，上皮钙黏素、LC3阳性细胞数低于空白组，上皮钙黏素、LC3-Ⅱ蛋白表达低于空白组，LC3-Ⅱ mRNA表达低于空白组，p62蛋白及其mRNA表达高于空白组（P＜0.05）。加味丹玄口康组自噬小体数量高于模型组，上皮钙黏素、LC3-Ⅱ阳性细胞数高于模型组，上皮钙黏素、LC3-Ⅱ蛋白表达高于模型组，LC3-Ⅱ mRNA表达高于模型组，p62蛋白及其mRNA表达低于模型组（P＜0.05）。结论加味丹玄口康可改善大鼠OSF，可能与其自噬水平一定程度升高有关。

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	贺珺1*　　张婷1*　　胡亮1　　潘世杰1　　邹红1　　连科荃1　　谭劲2　　胡兆勇1▲

关键词 ：口腔黏膜下纤维化, 自噬, 加味丹玄口康, 轻链3Ⅱ

Abstract：Objective To investigate the influence of Jiawei Danxuan Koukang on oral submucous fibrosis (OSF) and the level of autophagy in rats. Methods Twenty-four SPF male SD rats (180-200 g, 4-8 weeks old) were randomly divided into blank group, model group, and Jiawei Danxuan Koukang group, with eight rats in each group. In the model group and Jiawei Danxuan Koukang group, arecoline 0.2 ml (10 mg/ml) was injected into the bilateral buccal mucosa of the rats, once every two days, and a small toothbrush was used to brush the bilateral buccal mucosa of the rats every day for eight weeks. White lesions were observed in the buccal mucosa of the rats to determine the success of the model. After the model was successfully established, the rats in the Jiawei Danxuan Koukang group were treated with 8 ml/(kg·d) Jiawei Danxuan Koukang by gavage, and the blank group and the model group were treated with the same amount of 0.9% sodium chloride solution by gavage for four weeks. After the experiment, the bilateral buccal mucosa of the three groups were collected for hema- toxylin-eosin staining and Masson staining to observe the pathological changes and collagen fiber deposition, and the number of autophagosomes in the three groups was observed by transmission electron microscope. Immunohistochemical staining was used to detect the expression of E-cadherin and microtubule- associated protein light chain 3 (LC3) positive cells in the three groups. Western blot was used to detect the expression of E-cadherin, LC3-Ⅱ, and selective autophagy adaptor protein p62. The mRNA expression of LC3-Ⅱ and p62 was detected by RT-PCR. Results In the model group, the oral mucosal epithelium was significantly atrophed, the collagen fibers were significantly thickened and increased, the epithelial layer was thinner, and a large amount of collagen fibers were deposited in the submucosal layer. In the Jiawei Danxuan Koukang group, the oral mucosal epithelium was thicker, the collagen fibers were reduced, and the atrophy of the epithelial layer was alleviated. In model group, the number of autophagosomes was lower than that in blank group, the number of E-cadherin and LC3 positive cells were lower than those in blank group, the expression of E-cadherin and LC3-Ⅱ protein were lower than those in blank group, the expression of LC3-Ⅱ mRNA was lower than that in blank group, and the expression of p62 protein and its mRNA were higher than those in blank group (P＜0.05). In Jiawei Danxuan Koukang group, the number of autophagosomes was higher than that in model group, the expression of E-cadherin and LC3-Ⅱ protein were higher than those in model group, the expression of LC3-Ⅱ mRNA was higher than that in model group, and the expression of p62 protein and its mRNA were lower than those in model group (P＜0.05). Conclusion Jiawei Danxuan Koukang can improve OSF in rats, which may be related to the increased level of autophagy.

Key words： Oral submucous fibrosis Autophagy Jiawei Danxuan Koukang Light chain 3 Ⅱ

基金资助:国家自然科学基金资助项目（81874496）；
湖南省教育厅科学研究一般项目（20C1389）；
湖南省级大学生创新创业训练计划项目（S202110541057）。

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引用本文:

贺珺1*　　张婷1*　　胡亮1　　潘世杰1　　邹红1　　连科荃1　　谭劲2　　胡兆勇1▲. 加味丹玄口康对大鼠口腔黏膜下纤维化及其自噬水平的影响[J]. 中国医药导报, 2023, 20(24): 9-15.
HE Jun1* ZHANG Ting1* HU Liang1 PAN Shijie1 ZOU Hong1 LIAN Kequan1 TAN Jin2 HU Zhaoyong1▲. Influence of Jiawei Danxuan Koukang on oral submucosal fibrosis and the level of autophagy in rats. 中国医药导报, 2023, 20(24): 9-15.

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[1] 翦新春，高兴.口腔黏膜下纤维性变的病因、致病机理、诊断与治疗[J].口腔疾病防治，2021，29（4）：217-225.
[2] Shen YW，Shih YH，Fuh LJ，et al. Oral Submucous Fibrosis：A Review on Biomarkers，Pathogenic Mechanisms，and Treatments [J]. Int J Mol Sci，2020，21（19）：7231.
[3] 唐瞻贵，成雨熹.口腔黏膜下纤维性变临床诊治的研究进展[J].口腔医学研究，2022，38（8）：705-709.
[4] Cai X，Yao Z，Liu G，et al. Oral submucous fibrosis：A clinicopathological study of 674 cases in China [J]. Oral Pathol Med，2019，48（ 4）：321-325.
[5] Korolchuk VI，Menzies FM，Rubinsztein DC. Mechanisms of cross-talk between the ubiquitin-proteasome and autophagy- lysosome systems [J]. FEBS Lett，2010，584（7）：1393-1398.
[6] Migneault F，Hébert MJ. Autophagy，tissue repair，and fibrosis：a delicate balance [J]. Matrix Biol，2021，100-101：182- 196.
[7] Hill C，Li J，Liu D，et al. Autophagy inhibition-mediated epithelial-mesenchymal transition augments local myofibroblast differentiation in pulmonary fibrosis [J]. CDDIS，2019， 10（8）：591.
[8] Liang S，Wu YS，Li DY，et al. Autophagy and Renal Fibrosis [J]. Aging Dis，2022，13（3）：712-731.
[9] Zhang XL，Chen ZN，Huang QF，et al. Methyl Helicterate Inhibits Hepatic Stellate Cell Activation Through Modulation of Apoptosis and Autophagy [J]. Cell Physiol Biochem，2018， 51（2）：897-908.
[10] Li J，Zhao TT，Zhang P，et al. Autophagy mediates oral sub mucous fibrosis [J]. Exp Ther Med，2016，11（5）：1859- 1864.
[11] More CB，Jatti Patil D，Rao NR. Medicinal management of oral submucous fibrosis in the past decade- A systematic review [J]. Oral Biol Craniofac Res，2020，10（4）：552-568.
[12] 王宗康，肖艳波，刘寻，等.加味丹玄口康调控口腔黏膜下纤维化大鼠Wnt/β-catenin信号通路机制研究[J].陕西中医，2022，43（6）：687-690，774.
[13] Chiang MH，Chen PH，Chen YK，et al. Characterization of a Novel Dermal Fibrosis Model Induced by Areca Nut Extract that Mimics Oral Submucous Fibrosis [J]. PLoS One，2016，11（11）：e0166454.
[14] Maria S，Kamath VV，Satelur K，et al. Evaluation of transforming growth factor beta1 gene in oral submucous fibrosis induced in Sprague-Dawley rats by injections of areca nut and pan masala（commercial areca nut product）extracts [J]. Cancer Res Ther，2016，12（1）：379-385.
[15] 杨博，符梦凡，唐瞻贵.槟榔碱及机械刺激构建大鼠口腔黏膜下纤维化模型[J].华西口腔医学杂志，2019，37（3）：260-264.
[16] 李群，谭劲，刘寻等.环氧合酶-2、Runt相关转录因子3及生存素在口腔黏膜下纤维性变癌变中的表达[J].中国临床研究，2021，34（12）：1601-1604.
[17] Levine B，Kroemer G. Biological Functions of Autophagy Genes：A Disease Perspective [J]. Cell，2019，176（1/2）：11- 42.
[18] Dikic I，Elazar Z. Mechanism and medical implications of mammalian autophagy [J]. Nat Rev Mol Cell Biol，2018，19（6）：349-364.
[19] Kocaturk NM，Akkoc Y，Kig C，et al. Autophagy as a molecular target for cancer treatment [J]. Eur J Pharm Sci，2019， 134：116-137.
[20] Runwal G，Stamatakou E，Siddiqi FH，et al. LC3-positive structures are prominent in autophagy-deficient cells [J]. Sci Rep，2019，9（1）：10147.
[21] Jeong SJ，Zhang X，Rodriguez-Velez A，et al. p62/SQSTM1 and Selective Autophagy in Cardiometabolic Diseases [J]. Antioxid Redox Signal，2019，31（6）：458-471.
[22] Tao Y，Wang N，Qiu T，et al. The Role of Autophagy and NLRP3 Inflammasome in Liver Fibrosis [J]. Biomed Res Int，2020，2020：7269150.
[23] 李元聪.从痰瘀论治口腔黏膜下纤维化临证体会[J].湖南中医药大学学报，2018，38（7）：755-756.
[24] 李元聪，谭劲，文倩，等.口腔黏膜下纤维化中医临床诊疗方案[J].实用口腔医学杂志，2018，34（6）：838-840.
[25] 王雅莉，胡光，张倩，等.活血化瘀中药丹参药对的研究进展[J].重庆理工大学学报（自然科学），2020，34（3）：197-204.
[26] 郭锦材，谢辉.中医药治疗口腔黏膜下纤维化的临床研究进展[J].口腔医学研究，2019，35（5）：423-426.
[27] 魏佳涵，谭劲，胡淳.丹红注射液联合活血化瘀解毒法对口腔黏膜下纤维化的临床效果及对MCP-1、VEGF的影响[J].中国医药科学，2023，13（10）：4-7，19.
[28] Cao S，Chen S，Qiao X，et al. Protocatechualdehyde Rescues Oxygen-Glucose Deprivation/Reoxygenation-Induced Endothelial Cells Injury by Inducing Autophagy and Inhibiting Apoptosis via Regulation of SIRT1 [J]. Front Pharmacol，2022，13：846513.
[29] Shi X，Sun R，Zhao Y，et al. Promotion of autophagosome- lysosome fusion via salvianolic acid A-mediated SIRT1 up-regulation ameliorates alcoholic liver disease [J]. RSC Adv，2018，8（36）：20411-20422.
[30] Xu C，Ni S，Zhuang C，et al. Polysaccharide from Angelica sinensis attenuates SNP-induced apoptosis in osteoarthritis chondrocytes by inducing autophagy via the ERK1/2 pathway [J]. Arthritis Res Ther，2021，23（1）：47.
[31] Ye J，Lu S，Wang M，et al. Hydroxysafflor Yellow A Protects Against Myocardial Ischemia／Reperfusion Injury viaSuppressing NLRP3 Inflammasome and Activating Autophagy [J]. Front Pharmacol，2020，11：1170.