蛇床子-海螵蛸对白色念珠菌性阴道炎模型大鼠阴道黏膜修复的影响

Effect of the common cnidium fruit- cuttlebone on the repair of vaginal mucosa in rats with candida albicans vaginitis model

关键词:  蛇床子  海螵蛸  白色念珠菌  阴道黏膜

Key words:  common cnidium fruit  cuttlebone  candida albicans  vaginal mucosa

摘要:

[目的] 观察蛇床子-海螵蛸对白色念珠菌性阴道炎模型大鼠阴道黏膜修复的影响,比较蛇床子-海螵蛸和蛇床子治疗白色念珠菌性阴道炎模型大鼠阴道黏膜的作用差异。[方法] 取40只SPF级SD雌鼠(体质量180~220 g),并将大鼠随机分为空白组、模型组、蛇床子组、蛇床子-海螵蛸组,每组10只,建立白色念珠菌阴道炎(VVC)大鼠模型,空白组和模型组均采用生理盐水冲洗阴道,蛇床子组及蛇床子-海螵蛸组使用5 g生药/mL的发酵药物进行阴道灌洗治疗,每只每次阴道灌洗1 mL,每日1次,共灌洗10 d。采用pH试纸测定大鼠阴道pH值、苏木素-伊红(HE)染色观察大鼠阴道组织学病理、酶联免疫吸附测定法(ELISA)法检测大鼠阴道组织中白细胞酯酶(LE)、乙酰胺葡萄糖苷酶(OGA)蛋白水平及进行转录组学分析修复阴道黏膜机制。[结果] 成功建立VVC大鼠模型,与空白组比较,模型组大鼠阴道黏膜结构不完整,黏膜厚度降低、炎症浸润加剧,pH值及LE、OGA活性因子蛋白水平显著升高(P<0.01);与模型组相比,蛇床子-海螵蛸组及蛇床子组上述指标均下降(P<0.01),蛇床子-海螵蛸组上述指标均低于蛇床子组(P<0.05),且蛇床子-海螵蛸组炎症细胞浸润减少、阴道黏膜结构得到了一定恢复,黏膜厚度增加,而蛇床子组无明显的阴道黏膜修复作用;根据转录组结果可知,与模型组相比,蛇床子-海螵蛸发酵组差异基因一共523个,KEGG富集通路表达上调的主要为过氧化物酶体增殖物激活受体(PPAR)信号通路、细胞色素P450等。表达下调的主要为破骨细胞分化、白介素-17(IL-17)、肿瘤坏死因子(TNF)等信号通路。[结论] 蛇床子海螵蛸能够有效修复VVC模型大鼠阴道黏膜结构,能通过调节阴道pH值,改善阴道菌群,修复并保护阴道黏膜结构。蛇床子-海螵蛸可能是通过调控PPAR通路以及细胞P450酶,达到修复阴道黏膜的作用。

Abstract:

[Objective] To observe the effect of the common cnidium fruit-cuttlebone on the the repair of vaginal mucosa in rats with candida albicans vaginitis model and to compare the effects of common cnidium fruit-cuttlebone and common cnidium fruit on the vaginal mucosa in the candida albicans vaginitis model. [Methods] Forty Specific pathogen Free(SPF)-grade Sprague-Dawley(SD) female rats(body mass 180-220 g) were randomly divided into the blank group and model group,the common cnidium fruit group,the common cnidium fruit-cuttlebone group,10 rats in each group,to establish a rat model of candida albicans vaginitis(vulvovaginal candidiasis,VVC),saline was used for vaginal irrigation in both the blank group and model group,the vaginal irrigation treatment was carried out with a fermented drug at a concentration of 5 g of raw drug/mL in the common cnidium fruit group and common cnidium fruit-cuttlebone group. 1 mL of each vaginal irrigation was carried out once daily for 10 days. Determination of vaginal potentialofhydrogen(pH) in rats using pH paper,the histological pathology of rat vagina was observed by hematoxylin-eosin staining(HE) staining,the expression levels of leukocyte esterase(LE) and O-GlcNAcase(OGA) protein in rat vaginal tissue were detected by enzyme-linked immunosorbent assay(ELISA) and the mechanism of vaginal mucosa repair was analyzed by transcriptomics. [Results] The VVC rat model was successfully established,and compared with the blank group,the vaginal mucosa of the model group was structurally incomplete,the mucosal thickness was reduced,the inflammatory infiltration was intensified,and the pH value and the expression levels of LE and OGA activity factors were significantly increased(P<0.01);Compared to the model group,the above indexes were decreased in the common cnidium fruit-cuttlebone group and common cnidium fruit group(P<0.01),the common cnidium fruit-cuttlebone group is lower than common cnidium fruit group(P<0.05),and inflammatory cell infiltration was reduced,vaginal mucosal structure was restored to some extent,and mucosal thickness was increased in the common cnidium fruit-cuttlebone group,whereas the common cnidium fruit did not have a significant vaginal mucosal repair effect. According to the transcriptome results,there were 523 differential genes in the common cnidium fruit-cuttlebone group compared with the model group. The KEGG enrichment pathway was mainly up-regulated by the peroxisome proliferator-activated receptor(PPAR) signalling pathway and cytochrome P450. The main genes that were down-regulated were osteoclasts differentiation,interleukin-17(IL-17),tumor necrosis factor(TNF) and other signaling pathways. [Conclusion] The common cnidium fruit-cuttlebone can effectively repair the structure of vaginal mucosa in VVC model rats,can helps to repair and protect the structure of the vaginal mucosa by regulating vaginal pH and improving vaginal flora. The common cnidium fruit-cuttlebone may be able to repair the vaginal mucosa by regulating the PPAR pathway and cellular P450 enzymes.