尼非韦罗微乳热敏凝胶对小型猪阴道黏膜炎症刺激性的研究

摘要

Objective:To evaluate the inflammatory irritation of nifeviroc microemulsion thermosensitive gel (NFVR/ME-TISG) in the vaginal mucosa of mini-pigs.Methods:Nine female pigs were randomly divided into matrix gel group,NFVR/ME-TISG group (25 mg·d-1) and nonoxynol-9 (N-9) jelly group (25 mg·d-1).Twelve days after estradiol benzoate intramuscular injection,5 mL of gel was administered intravaginally once daily to the mini-pigs for 7 days.Cervicovaginal lavages (CVLs) were collected predose and at 24,72 h after the final administration.The numbers of neutrophilic granulocytes were analyzed by flow cytometry.Vaginal tissue was collected at 72 h after the final administration.Vaginal sections from each specimen were assessed after hematoxylin & eosin (HE) staining,and scored for the following histological features:epithelial ulceration/erosion,necrosis,cell infiltration,hemorrhage,congestion and edema.The CD45+ expression in the vaginal mucosa was examined by the immunohistochemical (IHC) assay.Results:Compared to the matrix gel group,the number of neutrophilic granulocytes of the N-9 jelly group was significantly increased (P ＜ 0.01).The vaginal mucosa of the N-9 jelly group showed obvious histopathological changes including epithelium loss,inflammation,bleeding and edema.CD45+ expression in the vaginal mucosa in N-9 jelly group was increased significantly (P ＜ 0.05).However,NFVR/METISG group showed no obvious changes.Conslusion:NFVR/ME-TISG did not induce the inflammatory irritation in the vaginal mocusa of mini-pigs,so they are promising to be developed into a topical microbicide.%目的:研究尼非韦罗微乳热敏凝胶(NFVR/ME-TISG)对小型猪阴道黏膜的炎症刺激性.方法:将9只雌性小型猪随机分为基质凝胶组、NFVR/ME-TISG组(25 mg·d-1)和壬苯醇醚-6(N-9)凝胶组(25 mg·d-1).肌注苯甲酸雌二醇后12d,开始阴道给药5 mL·d-1,连续7d,给药前和末次给药后24与72 h,收集阴道灌洗液(cervicovaginal lavages,CVL),流式细胞仪检测CVL中的中性粒细胞,末次给药后72 h收集阴道组织,苏木素伊红染色法(hematoxylin&eosin,HE)评价小型猪阴道黏膜组织溃疡、坏死、细胞浸润、出血、充血、水肿等指标,免疫组织化学法(immuno-histochemical,IHC)检测阴道黏膜组织中CD45+表达的分布.结果:与基质凝胶组相比,N-9凝胶组CVL中炎性细胞的中性粒细胞数目明显增加(P＜0.01),阴道黏膜出现上皮局部脱落、炎性细胞增多、出血和水肿,CD45+累积光密度(integrated optical density,IOD)值(33.95±6.69)显著增高(P＜0.05);而NFVR/ME-TISG组未见明显变化.结论:NFVR/ME-TISG未明显引起小型猪阴道黏膜的炎症刺激性,安全性较好,有望作为阴道外用杀微生物剂候选物进一步研发.