槟榔碱及机械刺激构建大鼠口腔黏膜下纤维化模型

doi:10.7518/hxkq.2019.03.006

摘要/Abstract

摘要：

目的利用槟榔碱和机械刺激构建Sprague-Dawley（SD）大鼠口腔黏膜下纤维化（OSF）模型。方法采用两因素析因实验设计将48只大鼠分为8个组,每组6只。分别用不同浓度（0、0.5、2、8 mg·mL ^-1）槟榔碱涂擦及机械刺激 (有或无毛刷涂擦)。处理16周后测量开口度;取局部颊黏膜行苏木精-伊红（HE）染色观察病理变化;并检测组织内Ⅲ型胶原、转化生长因子-β1（TGF-β1）和γ干扰素（IFN-γ）的表达情况。结果 2和8 mg·mL ^-1（中、高）浓度槟榔碱处理16周,颊黏膜出现了典型的OSF病理特征;开口度显著减小并且Ⅲ型胶原、TGF-β1表达显著增加（P<0.05）。机械刺激虽可导致黏膜Ⅲ型胶原、TGF-β1和IFN-γ表达增高（P<0.05）,但无病理学改变,开口度改变不显著。结论中、高浓度槟榔碱有致OSF作用;机械刺激无法导致大鼠OSF的发生。

关键词: 槟榔碱, 机械刺激, 口腔黏膜下纤维化, 动物模型

Abstract:

Objective The aim of this study was to induce oral submucous fibrosis (OSF) in Sprague-Dawley(SD) rat models by arecoline and mechanical stimulation. Methods Two factors factorial design was used to divide 48 rats into 8 groups (n=6). Different concentrations of arecoline (0, 0.5, 2, and 8 mg·mL ^-1) and mechanical stimulation (with or without brush) were treated. After 16 weeks of treatment, the mouth opening was measured, the pathological changes of the buccal mucosa were observed, and the expressions of type Ⅲ collagen, transforming growth factor β1 (TGF-β1), and interferon-γ (IFN-γ) were detected. Results In rats with moderate and high concentrations of arecoline, typical OSF pathological features were observed in the buccal mucosa, the mouth openings were significantly reduced, and the expression levels of type Ⅲ collagen and TGF-β1 were significantly increased (P<0.05). Although mechanical stimulation can increase the three indexes of mucosa (P<0.05), no pathological change and difference in the mouth opening was observed (P>0.05). Conclusion Moderate and high concentrations of arecoline can induce OSF in SD rats, but mechanical stimulation cannot induce OSF.

Key words: arecoline, mechanical stimulation, oral submucous fibrosis, animal model

中图分类号:

R781.5

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.

Bo Yang,Mengfan Fu,Zhangui Tang. Rat model with oral submucous fibrosis induced by arecoline and mechanical stimulation[J]. West China Journal of Stomatology, 2019, 37(3): 260-264.

图/表 7

表 1

图 1

表 2

表 3

表 4

图 2

图 3

参考文献 18

[1]	Arakeri G, Rai KK, Hunasgi S , et al. Oral submucous fibrosis: an update on current theories of pathogenesis[J]. J Oral Pathol Med, 2017,46(6):406-412. doi: 10.1111/jop.2017.46.issue-6 URL
[2]	Caro V, Sutera FM, Giannola LI . In situ delivery of corticosteroids for treatment of oral diseases[J]. Ther Deliv, 2017,8(10):899-914. doi: 10.4155/tde-2017-0055 URL
[3]	Yang YH, Lee HY, Tung S , et al. Epidemiological survey of oral submucous fibrosis and leukoplakia in aborigines of Taiwan[J]. J Oral Pathol Med, 2001,30(4):213-219. doi: 10.1034/j.1600-0714.2001.300404.x URL
[4]	Lee CH, Ko AMS, Warnakulasuriya S , et al. Intercountry prevalences and practices of betel-quid use in south, southeast and eastern Asia regions and associated oral preneoplastic disorders: an international collaborative study by Asian betel-quid consortium of south and east Asia[J]. Int J Cancer, 2011,129(7):1741-1751. doi: 10.1002/ijc.v129.7 URL
[5]	Tang JG, Jian XF, Gao ML , et al. Epidemiological survey of oral submucous fibrosis in Xiangtan City, Hunan Province, China[J]. Community Dent Oral Epidemiol, 1997,25(2):177-180. doi: 10.1111/com.1997.25.issue-2 URL
[6]	Rajendran R, Raju GK, Nair SM , et al. Prevalence of oral submucous fibrosis in the high natural radiation belt of Kerala, south India[J]. Bull World Health Organ, 1992,70(6):783-789.
[7]	Sirsat SM, Khanolkar VR . Submucous fibrosis of the palate in diet-preconditioned Wistar rats. Induction by local painting of capsaicin: an optical and electron microscopic study[J]. Arch Pathol, 1960,70:171-179.
[8]	Sumeth Perera MW, Gunasinghe D, Perera PA , et al. Development of an in vivo mouse model to study oral submucous fibrosis[J]. J Oral Pathol Med, 2007,36(5):273-280. doi: 10.1111/jop.2007.36.issue-5 URL
[9]	Zhang SS, Gong ZJ, Xiong W , et al. A rat model of oral submucous fibrosis induced by bleomycin[J]. Oral Surg Oral Med Oral Pathol Oral Radiol, 2016,122(2):216-223. doi: 10.1016/j.oooo.2015.07.042 URL
[10]	Wen QT, Wang T, Yu DH , et al. Development of a mouse model of arecoline-induced oral mucosal fibrosis[J]. Asian Pac J Trop Med, 2017,10(12):1177-1184. doi: 10.1016/j.apjtm.2017.10.026 URL
[11]	Sharma A, Kumar R, Johar N , et al. Oral submucous fibrosis: an etiological dilemma[J]. J Exp Ther Oncol, 2017,12(2):163-166. URL pmid: 29161786
[12]	Shanbhag VK . Medical management of oral submucous fibrosis[J]. Asia Pac J Oncol Nurs, 2015,2(1):51. doi: 10.4103/2347-5625.143766 URL
[13]	Razi A, Iqbal A, ul Wahab N . Topical application of tocopherol with corticosteroid in reducing the early signs and symptoms of oral submucous fibrosis patients[J]. Annals Abbasi Shaheed Hospital Karachi Med Dent College, 2016,21(2):100-105.
[14]	Utsunomiya H, Tilakaratne WM, Oshiro K , et al. Extracellular matrix remodeling in oral submucous fibrosis: its stage-specific modes revealed by immunohistochemistry and in situ hybridization[J]. J Oral Pathol Med, 2005,34(8):498-507. doi: 10.1111/jop.2005.34.issue-8 URL
[15]	谢辉, 刘健, 凌天牖 . 细胞因子IL-6, TGF-β1在口腔黏膜下纤维性变中的表达[J]. 临床口腔医学杂志, 2012,28(10):592-594.
	Xie H, Liu J, Ling TY . Expressions of chemokine IL-6, TGF-β1 in oral submucous fibrosis[J]. J Clin Stomatol, 2012,28(10):592-594.
[16]	Zhang ZR, Chen LY, Qi HY , et al. Expression and clinical significance of periostin in oral lichen planus[J]. Exp Ther Med, 2018,15(6):5141-5147.
[17]	Stoppel WL, Kaplan DL, Black LD . Electrical and mechanical stimulation of cardiac cells and tissue constructs[J]. Adv Drug Deliv Rev, 2016,96(1):135-155. doi: 10.1016/j.addr.2015.07.009 URL
[18]	王涛, 温琦涛, 于大海 , 等. 一种口腔黏膜下纤维化小鼠模型的构建方法: 中国, CN106511345A[P]. [2017-03-22].
	Wang T, Wen QT, Yu DH , et al. Establishment of a mouse model of oral submucousfibrosis: China, CN106511345A[P], [2017-03-22].

组别	开口度/cm	Ⅲ型胶原	TGF-β1	IFN-γ
A	2.203±0.115	0.08±0.06	0.18±0.05	0.16±0.08
B	2.191±0.069	0.11±0.06	0.19±0.02	0.17±0.09
C	2.000±0.104	0.12±0.06	0.25±0.04	0.12±0.09
D	1.942±0.073	0.14±0.08	0.24±0.03	0.15±0.07
E	2.190±0.084	0.21±0.06	0.28±0.03	0.51±0.10
F	2.167±0.070	0.23±0.09	0.29±0.05	0.41±0.09
G	2.003±0.156	0.38±0.08	0.39±0.06	0.41±0.11
H	1.978±0.078	0.41±0.08	0.39±0.06	0.38±0.09

源	开口度		Ⅲ型胶原		TGF-β1		IFN-γ
源	F值	P值	F值	P值	F值	P值	F值	P值
槟榔碱浓度（a）	18.591	0.000^*	82.144	0.000^*	13.390	0.000^*	1.441	0.245
机械刺激（b）	0.002	0.965	9.244	0.000^*	87.125	0.000^*	109.539	0.000^*
槟榔碱浓度×机械刺激（a×b）	0.232	0.873	3.016	0.041^*	1.172	0.333	1.035	0.388

槟榔碱浓度（I）/	槟榔碱浓度（J）/	开口度		Ⅲ型胶原		TGF-β1
（mg·mL^-1）	（mg·mL^-1）	平均值差值（I-J）	P值	平均值差值（I-J）	P值	平均值差值（I-J）	P值
0	0.5	0.018 3	0.647	-0.026 7	0.384	-0.005 8	0.757
	2	0.195 0	0.000^*	-0.111 7	0.001^*	-0.090 0	0.000^*
	8	0.237 5	0.000^*	-0.135 0	0.000^*	-0.083 3	0.000^*
0.5	2	0.176 7	0.000^*	-0.085 0	0.008^*	-0.084 2	0.000^*
	8	0.219 2	0.000^*	-0.108 3	0.001^*	-0.077 5	0.000^*
2	8	0.042 5	0.291	-0.023 3	0.446	0.006 7	0.724