壬苯醇醚-9诱导小型猪阴道黏膜免疫炎症反应模型的建立

doi:10.3969/j.issn.1674-5817.2016.02.001

实验动物与比较医学 ›› 2016, Vol. 36 ›› Issue (2): 81-86.DOI: 10.3969/j.issn.1674-5817.2016.02.001

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壬苯醇醚-9诱导小型猪阴道黏膜免疫炎症反应模型的建立

黄超¹, 弓艺涓¹, 田芳¹, 王玉柱¹, 孙冰^1,2, 夏敏杰¹, 职瑞娜^1,2, 丁训诚¹, 周新初³, 李卫华¹

1.上海市计划生育科学研究所, 国家计划生育药具重点实验室, 上海 200032;
2.复旦大学公共卫生学院, 上海 200032;
3.上海交通大学农业与生物学院, 上海, 201101

收稿日期:2015-12-07 出版日期:2016-04-25 发布日期:2016-04-25
作者简介:黄超, 男, 本科, 主要从事新药研发及生殖药理评价研究。E-mail: 553026696@qq.com; 同第一作者, 弓艺涓, 女, 硕士, 主要从事药物研发。E-mail: gongyijuan@126.com
基金资助:
国家科技重大专项(2013ZX10001006-003-002); 国家科技重大专项(2012ZX10001007-009-003); 十二五国家科技支撑计划(2012BAI31B04); 上海市科委项目(13431900800)

Establishment of Vaginal Mucosal Immuno-inflammatory Response Model in Minipig

HUANG Chao¹, GONG Yi-juan¹, TIAN Fang¹, WANG Yu-zhu¹, SUN Bing^1,2, ZHI Rui-na^1,2, IA Min-jie¹, DING Xun-cheng¹, ZHOU Xin-chu³, LI Wei-hua¹

1. State Key Lab of Family Planning Drugs and Devices, Shanghai Institute of Planned Parenthood Research, Shanghai 200032, China;
2. School of Public Health, Fudan University, Shanghai 200032;
3. Shanghai Jiao Tong University School of Agriculture and Biology, Shanghai 201101, China

Received:2015-12-07 Online:2016-04-25 Published:2016-04-25

摘要/Abstract

摘要： 目的探索建立广西巴马小型猪阴道黏膜免疫炎症反应模型的方法。方法将6只雌性小型猪随机分为基质对照组和壬苯醇醚-9(N-9)模型组(40 mg/mL)(每组3只)。肌肉注射苯甲酸雌二醇12 d后,各组阴道给药5 mL/d,连续7 d。造模后取阴道黏膜组织进行病理检查,并根据上皮溃疡、坏死、细胞浸润、出血、充血、水肿等指标进行病理评分; 造模后24 h与72 h,采集阴道灌流液(CVL), 采用流式细胞术对CVL沉淀细胞中的中性粒细胞比例和CD45⁺细胞百分率进行分析; 免疫组织化学法(IHC)检测阴道黏膜组织中CD45⁺细胞的表达及分布。结果组织病理学观察显示,N-9模型组阴道黏膜上皮损伤明显,出现不同程度的溃疡、坏死、炎性浸润、出血、水肿等。病理评分结果显示,N-9模型组病理总分值(18.0±3.2)与基质对照组(3.3±0.2)相比明显升高(P<0.01)。N-9 模型组CVL的中性粒细胞比例为41.70%±1.63% (24 h)和51.17%±3.81%(72 h), CD45⁺细胞比例为58.5%±4.73%(24 h)和88.0%±3.57 %(72 h),与基质对照组相比均显著增高(P<0.01)。N-9模型组阴道黏膜CD45⁺细胞表达明显增多,累积光密度值为33.95±6.69,显著高于基质对照组(23.42±3.30)(P<0.05)。结论阴道连续给予N-9凝胶7 d可以诱发小型猪阴道黏膜明显的炎性反应,符合阴道黏膜免疫炎症反应模型的特征。

关键词: 小型猪, 壬苯醇醚-9(N-9), 病理组织学, 阴道灌流液(CVL), 免疫炎症反应, 白细胞共同抗原CD45

Abstract: Objective To establish the vaginal mucosal immuno-inflammatory model response of minipig. Methods Six female minipigs were randomly divided into the control and Nonoxynol-9 (N-9) model group (40 mg/mL). After 12 days of estradiol benzoate intramuscular injection, the test drugs (N-9) were administered through vagina to minipigs at 5 mL/d for 7 consecutive days. On the 7th day, histopathological examination of vaginal mucosa was undertaken and the pathological scores were calculated based on histological indexes including epithelial ulceration/erosion, necrosis, cell infiltration, hemorrhage, congestion and edema. Cervicovaginal lavage (CVL) was collected at 24 h and 72 h after drug administration. The percentage of neutrophilic granulocytes and CD45⁺ cells in the CVL deposited cells were detected by flow cytometry. The expression and location of CD45⁺ cells in the vaginal mucosa were assessed by using immuno-histochemical (IHC) assay. Results Histopathological examination showed obvious lesions including inflammation, bleeding and edema in the vaginal epithelium of the N-9 group. Histological score of the N-9 group (18.0±3.2) was significantly higher than that of the control (3.3±0.2) (P<0.01). The percentage of neutrophilic granulocytes in the CVL cells of N-9 group was (41.70±1.63)% (24 h) and (51.17±3.81)% (72 h), and the percentage of CD45⁺ cells in the CVL cells of N-9 group was (58.5±4.73) % (24 h) and (88.0±3.57)% (72 h), both showed significant increase compared to that of the control (P<0.01). CD45⁺ cell expression increased significantly in the N-9 group. The IOD value of N-9 group was 33.95±6.69, which was significantly higher than that of the control group (28.42±8.96) (P<0.05). Conclusion The vaginal mucosal immuno-inflammatory response model of mini-pigs could be induced by giving 40 mg/mL N-9 gel to the mini-pigs through vagina for 7 consecutive days.

Key words: Minipig, Nonoxynol-9 (N-9), Histopathology, Cervicovaginal lavage(CVL), Immuno-inflammation response, Leuckocyte common antigen-CD45

中图分类号:

Q95-33

黄超, 弓艺涓, 田芳, 王玉柱, 孙冰, 夏敏杰, 职瑞娜, 丁训诚, 周新初, 李卫华. 壬苯醇醚-9诱导小型猪阴道黏膜免疫炎症反应模型的建立[J]. 实验动物与比较医学, 2016, 36(2): 81-86.

HUANG Chao, GONG Yi-juan, TIAN Fang, WANG Yu-zhu, SUN Bing, ZHI Rui-na, IA Min-jie, DING Xun-cheng, ZHOU Xin-chu, LI Wei-hua. Establishment of Vaginal Mucosal Immuno-inflammatory Response Model in Minipig[J]. Laboratory Animal and Comparative Medicine, 2016, 36(2): 81-86.

参考文献

[1] Thompson IO, van der Bijl P, van Wyk CW, et al. A comparative light-microscopic, electron-microscopic and chemical study of human vaginal and buccal epithelium[J]. Arch Oral Biol, 2001, 46(12):1091-1098.
[2] Davis CC, Kremer MJ, Schlievert PM, et al.Penetration of toxic shock syndrome toxin-1 across porcine vaginal mucosa ex vivo: permeability characteristics, toxin distribution, and tissue damage[J]. Am J Obstet Gynecol, 2003, 189(6):1785-1791.
[3] D’Cruz OJ, Erbeck D, Uckun FM. A study of the potential of the pig as a model for the vaginal irritancy of benzalkonium chloride in comparison to the nonirritant microbicide PHI-443 and the spermicide vanadocene dithiocarbamate[J]. Toxicol Pathol, 2005, 33(4):465-476.
[4] Wikinson D, Tholandi M, Ramjea G, et al.Nonoxynol-9 Apermicide for prevention of vaginally acquired HIV and other sexually transmitted infections: sgstematic review and meta-analgsis of randomized controlled trials including more than 5000 women[J]. Lancet Infect Dis, 2002, 2(1):613-617.
[5] Obiero J, Mwethera PG, Hussey GD, et al.Vaginal microbicides for reducing the risk of sexual acquisition of HIV infection in women: systematic review and meta- analysis[J]. BMC Infect Dis, 2012, 12(5):289-310.
[6] Patton DL, Cosgrove SY, Paul KJ.A summary of preclinical topical microbicide vaginal safety and chlamydial efficacy evaluations in a pigtailed macaque model[J]. Sex Transm Dis,2008, 35(10):889-897.
[7] Peuster M, Fink C, Reckers J, et al.Assessment of subacute inflammatory and proliferative response to coronary stenting in a porcine model by local gene expression studies and histomorphometry[J]. Biomaterials, 2004, 25(6):957-963.
[8] Catalone BJ, Kislr-Catalone TM, Budgeon LR, et al.Mouse model of cericoraginal toxicity and inflammation for preclinical eraluation of topical vaginal microbicides[J]. Antimicrob Agents Chemother, 2004, 48(5):1837-1847.
[9] Lard-Whiteford SL, Matecxcka D, O'Rear JJ, et al. Recommendations for the nonclinical development of topical microbicides for prevention of HIV transmission: an update[J]. J Acquir Immune Defic Syndr, 2004, 36(1):541-552.
[10] D’Cruz OJ, Uckun FM. Preclinical evaluation of a dual-acting microbicidal prodrug WHI-07 in combination with vanadocene dithiocarbamate in the female reproductive tract of rabbit, pig, and cat[J]. Toxicol Pathol, 2007, 35(7):910-927.
[11] Doncel GF, Chandra N, Fichorova RN.Preclinical assessment of the proinflammatory potential of microbicide candidates[J]. J Acquir Immune Defic Syndr, 2004, 37(Suppl 3):S174-S180.
[12] Zalenskaya IA, Cerocchi OG, Joseph T, Donaghay MA, et al.Increased cox-2 expression in human vaginal epithelial cells exposed to nonoxynol-9, a vaginal contraceptive microbicide that failed to protect women from HIV-1 infection[J]. Am J Reprod Immunol, 2011, 65(2):569-577.
[13] Cummins JE, Doncel GF.Biomarkers of cervicovaginal inflammation for the assessment of microbicide safety[J]. Sex Transm Dis, 2009, 36(3 Suppl):S84-S91.

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壬苯醇醚-9诱导小型猪阴道黏膜免疫炎症反应模型的建立

Establishment of Vaginal Mucosal Immuno-inflammatory Response Model in Minipig

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