白念珠菌口腔黏膜感染小鼠模型的建立及评估

摘要/Abstract

摘要： ［摘要］目的建立白念珠菌口腔黏膜感染ICR小鼠模型，动态观察口腔黏膜及相应的全身感染情况，并对变化情况进行初步分析。方法将白念珠菌接种于免疫功能低下小鼠口腔，采用肉眼观察口腔舌背病损改变、口腔内白念珠菌菌量检测、组织病理学等方法评估口腔内黏膜感染情况，检测体重、载菌量（肾、肝、下降结肠内粪便）评估全身感染情况。观察1周，并记录数据。空白对照组为接种生理盐水的免疫功能低下组。结果 ①小鼠接种白念珠菌后第1天口腔白念珠菌菌量较低，随后迅速增长，3~7 d内趋于稳定，稳定于106~107CFU/mL。同时口腔舌背病损也出现类似趋势，第1天未见明显伪膜，3~7 d内可见舌背伪膜存在。②病理切片PAS染色显示接种后3~5 d内白念珠菌形成菌丝侵入并破坏上皮，到接种第7天时，黏膜上皮多见酵母细胞。③小鼠接种白念珠菌后体重逐渐下降。粪便载菌量逐渐上升，第5天达到最大，但肾、肝无白念珠菌感染。空白组未见白念珠菌感染。结论通过在ICR小鼠口腔内接种白念珠菌可以建立白念珠菌口腔黏膜感染动物模型。接种后白念珠菌感染程度在1周内存在变化。动物实验应根据感染状况选择合适的研究时段。

关键词: 【关键词】白念珠菌, 小鼠口腔白念珠菌病, 黏膜感染

Abstract: Abstract：Objective To establish a murine oral mucosal candidiasis model, observe oral mucosal infection and examine the corresponding systemic infectiondynamically, and to analyzethe changes of the infection. MethodsImmuno-suppressed mice were inoculated with C.albicansinfectionusing the oral swabs. Lesions in the murine tongue were observed using macroscopic observation. The viable cells of the C. albicans in the oral cavity were calculated and the histological study of the murine tongues was analyzed to determine C. albicans infections of the oral mucosal. Weight and fungal burdens in the kidney, liver and feces from the descending colon were examined to assess the severity of the systemic infections. The above indicators in the infected mice were examined and recorded for a week after inoculation. In the blank group immunosuppressed mice were swabbed with saline. Results①The number of viable C. albicans cells in the oral cavity was low on the 1st day after inoculation，then rapidly grew and tended to be stable between the range of 106-107CFU / ml from day 3 to day 7 after inoculation . Oral tongue lesions had a similar trend. There was no obvious pseudomembranous on day 1 and visible tongue dorsal pseudomembranous could be seen from day 3 to day 7 after inoculation .②Histopathological sections stained by PAS revealed that C. albicans had developed a mycelial form, penetrated into the mucosal epithelium and destroyed the epithelium within 3-5 days after inoculation. There were visible yeast cells on the mucosal epithelium on day 7 after inoculation.③The weight of the mice gradually decreased after inoculation. Candida cells that recovered from feces gradually increased and reached to the highest level on the 5th day afterinoculation, but in kidneys and livers , there were no viable Candida cells . The blank group was not infected with C. albicans . Conclusions：Inoculating C. albicans in the oral cavity of mice using oral swabs can establish a murine oral mucosal candidiasis model. The oral candidiasis of the infected mice has changes in a week. The appropriate time of animal experiments should have been chosen depending on the infection status

Key words: 【Key words】Candida albicans, Murine oral candidiasis, Mucosal infection

中图分类号:

R780.2

李成蹊郑林霞左露露黄云生陈圣琰魏昕. 白念珠菌口腔黏膜感染小鼠模型的建立及评估[J]. , 2018, 38(5): 394-398.

参考文献

[1]Campisi G, Panzarella V, Matranga D, et al.Risk factors of oral candidosis: a twofold approach of study by fuzzy logic and traditional statistic[J].Arch Oral Biol, 2008, 53(4):388-397
[2]Soysa NS, Samaranayake LP, Ellepola AN.Diabetes mellitus as a contributory factor in oral candidosis[J].Diabet Med, 2006, 23(5):455-459
[3]Weerasuriya N, Snape J.Oesophageal candidiasis in elderly patients risk factors,prevention and management[J].Drugs Aging, 2008, 25(2):119-130
[4]Soysa NS, Samaranayake LP, Ellepola AN.Antimicrobials as a contributory factor in oral candidosis--a brief overview[J].Oral Dis, 2008, 14(2):138-143
[5]Samaranayake YH, Samaranayake LP.Experimental oral candidiasis in animalmodels[J].Clin Microbiol Rev, 2001, 14(2):398-429
[6]Junqueira JC, Vilela SF, Rossoni RD, et al.Oral colonization by yeasts in HIV-positive patientsin Brazil[J].Rev Inst Med Trop Sao Paulo, 2012, 54(1):17-24
[7]Badiee P, Alborzi A, Davarpanah MA, etal.Distributions and antifungal susceptibility of Candida species from mucosal sites in HIV positive patients[J].Arch Iran Med, 2010, 13(4):282-287
[8]卫彦, 陆慧君, 张新成, 等.白念珠菌对唑类抗真菌药物耐药机制的研究进展[J].口腔医学, 2005, 25(5):311-313
[9] Costa AC, Pereira CA, Junqueira JC, etal.Recent mouse and rat methods for the study of experimental oral candidiasis[J]. Virulence, 2013, 4:(5) 391-399.
[10]沈佳娜, 张钰, 黄韧，等.小鼠念珠菌感染模型和抗感染免疫[J].中国实验动物学报, 2008, 16(3):233-237
[11]Takakura N, Sato Y, Ishibashi H, et al.A novel murine model of oral candidiasis with local symptoms characteristic of oral thrush[J].Microbiol Immunol, 2003, 47(5):321-326
[12]Solis NV, Filler SG.Mouse model of oropharyngeal candidiasis[J].Nat Protoc, 2012, 7(4):637-642
[13]Dangi YS, Soni ML, Namdeo KP.Oral candidiasis: A review[J].International Journal of Pharmacy & Pharmaceutical Sciences, 2010, 2(4):36-41
[14]Bader T, Schr?ppel K, Bentink S, et al.Role of calcineurin in stress resistance,morphogenesis,and virulence of a Candida albicans wild-type strain[J].Infect Immun, 2006, 74(7):4366-4369
[15]Wang L, Wang C, Mei H, et al.Combination of Estrogen and Immunosuppressive Agents to Establish a Mouse Model of Candidiasis with Concurrent Oral and Vaginal Mucosal Infection[J].Mycopathologia, 2016, 181(1-2):29-39
[16]Okada M, Hisajima T, Ishibashi H, et al.Pathological analysis of the Candida albicans-infected tongue tissues of a murine oral candidiasis model in the early infection stage[J].Arch Oral Biol, 2013, 58(4):444-450
[17]罗银珠, 潘金春, 何丽芳, 等.白色念珠菌经口感染小鼠建立系统性感染模型[J].中国实验动物学报, 2016, 24(6):591-595
[18] 杨连娟.IL-22和TNF-α在口腔粘膜抗白念珠菌感染固有免疫中的协同作用[D].上海第二军医大学，2013.
[19]de Campos Rasteiro VM, da Costa AC, Araujo CF, et al.Essential oil of Melaleuca alternifolia for the treatment of oral candidiasis induced in an immunosuppressed mouse model[J].BMC Complement Altern Med, 2014, 14(1):1-10
[20] Freire F, Ferraresi C, Jorge AO, et al.Photodynamic therapy of oral Candida infection in a mouse model. [J].J Photochem Photobiol B. 2016, 159: 161-168.
[21]Hisajima T, Maruyama N, Tanabe Y, et al.Protective effects of farnesol againstoral candidiasis in mice[J].Microbiol Immunol, 2008, 52(7):327-333
[22]Junqueira JC.Models hosts for the study of oral candidiasis[J].Adv Exp Med Biol, 2012, 7(10):95-105