论文部分内容阅读
目的采用优化家兔阴道刺激模型评价乙酸阴道热敏凝胶剂(acetic acid vaginal thermosensitive in situ gel,Acid-TISG)对家兔阴道黏膜的免疫炎症毒性。方法将24只♀家兔随机分为空白凝胶组、Acid-TISG组(12.11 mg·m L~(-1))、壬苯醇醚(nonoxynol-9,N-9)凝胶组(5.6 mg·m L~(-1)),阴道给予3 m L凝胶,每天1次,连续10 d,给药前和给药后24 h收集家兔阴道灌洗液(cervicovaginal lavages,CVLs),检测CVL中白介素6(interleukin-6,IL-6)和肿瘤坏死因子α(tumor necrosis factorα,TNF-α)的含量。家兔处死后取阴道组织做病理学检查,HE染色观察家兔阴道黏膜组织损伤并进行评分。运用免疫组化技术检测阴道黏膜内白细胞共同抗原-CD45RB和核增殖抗原(proliferating cell nuclear antigen,PCNA)的表达。结果与空白凝胶组相比,Acid-TISG组黏膜上皮完整,无明显炎症浸润、充血和水肿,N-9凝胶组黏膜上皮出现脱落,固有层炎性细胞明显增多(P<0.05);空白凝胶组和Acid-TISG组阴道黏膜无明显CD45RB阳性表达,N-9凝胶组阴道黏膜固有层出现较多的CD45RB阳性表达细胞。与空白凝胶组相比,Acid-TISG组的阴道黏膜PCNA表达增多,但无显著性差异,N-9凝胶组的阴道黏膜PCNA阳性表达细胞明显增多(P<0.05)。与给药前相比,给药后24 h时CVL中IL-6和TNF-α含量在空白凝胶组和Acid-TISG组无明显改变,N-9凝胶组明显升高(P<0.05)。结论 Acid-TISG对家兔阴道黏膜无明显免疫炎症毒性,优化的家兔阴道刺激模型可用来评价生殖道外用药物对阴道黏膜的免疫炎症毒性。
Objective To evaluate the immune-inflammatory cytotoxicity of acetic acid vaginal thermosensitive in situ gel (Acid-TISG) on vaginal mucosa in rabbits. Methods Twenty-four rabbits were randomly divided into blank gel group, Acid-TISG group (12.11 mg · mL -1), nonoxynol-9 (5.6 mg · mL -1) The vaginal lavages (cervicovaginal lavages, CVLs) were collected before vaginal administration and 3 h after vaginal administration once daily for 10 days. The levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in CVL were measured. Vaginal tissues were taken from the rabbits for histopathology examination. The lesions of the vaginal mucosa were observed by HE staining and scored. Immunohistochemistry was used to detect the expression of CD45RB and proliferating cell nuclear antigen (PCNA) in the vaginal mucosa. Results Compared with the blank gel group, the mucosal epithelium of Acid-TISG group was intact with no inflammatory infiltration, congestion and edema. The mucosal epithelium of N-9 gel group was exfoliated and the number of inflammatory cells in lamina propria was significantly increased (P <0.05). There was no significant positive expression of CD45RB in the vaginal mucosa of the blank gel group and Acid-TISG group, while more CD45RB positive cells appeared in the lamina propria of N-9 gel group. Compared with blank gel group, the expression of PCNA in Acid-TISG group increased, but there was no significant difference. The positive expression of PCNA in vaginal mucosa of N-9 gel group was significantly increased (P <0.05). The levels of IL-6 and TNF-α in CVL were not changed in the blank group and Acid-TISG group at 24 hours after administration, but were significantly increased in the N-9 gel group (P <0.05) ). Conclusion Acid-TISG has no obvious immunological and inflammatory cytotoxicity on the vaginal mucosa in rabbits. The optimized vaginal stimulation model in rabbits can be used to evaluate the immune and inflammatory cytotoxicity of vaginal mucosa.