白头翁汤正丁醇提取物对外阴阴道念珠菌病小鼠阴道黏膜中性粒细胞趋化的影响

Effect of butyl alcohol extract of Baitouweng Decoction on vaginal mucosal neutrophil chemotaxis in vulvovaginal candidiasis mice

该文探讨白头翁汤正丁醇提取物(butyl alcohol extract of Baitouweng Decoction,BAEB)对外阴阴道念珠菌病(vulvovaginal candidiasis,VVC)小鼠阴道黏膜中性粒细胞趋化的影响。将SPF级雌性昆明小鼠72只,随机分为正常组,模型组,氟康唑组,BAEB低、中、高剂量组,皮下注射苯甲酸雌二醇致假发情期,再阴道接种白念珠菌2×106 CFU·mL-1,药物治疗7 d。革兰染色法观察小鼠阴道分泌物中白念珠菌形态;平板法检测小鼠阴道真菌载荷;HE染色法观察小鼠阴道组织病理情况;酶联免疫吸附(ELISA)法检测小鼠灌洗液中LDH,IL-6,TNF-α水平;巴氏染色法观察阴道灌洗液中性粒细胞并计数;ELISA法检测阴道黏膜趋化因子中IL-8和MIP-2水平;qRT-PCR法检测小鼠阴道黏膜中IL-8和MIP-2的mRNA水平。结果表明,与正常组相比,VVC模型组阴道内有大量菌丝生成且大量的真菌载荷,阴道黏膜层结构完全破坏,中性粒细胞数量增多,IL-8和MIP-2的蛋白和mRNA水平上调。经BAEB治疗,与模型组相比,治疗组小鼠阴道内菌丝减少,真菌载荷量降低,黏膜损伤有所改善,炎症因子含量降低,IL-8和MIP-2的蛋白和mRNA水平均下调。上述结果提示,BAEB可能通过抑制趋化因子以阻断中性粒细胞向阴道腔的募集而发挥治疗VVC的作用。

To investigate the effects of butyl alcohol extract of Baitouweng Decoction(BAEB)on neutrophil chemotaxis in vaginal mucosa of mice with vulvovaginal candidiasis(VVC).Seventy-two SPF female Kunming mice were randomly divided into normal control group,model group,fluconazole group,BAEB low-dose group,middle-dose group and high-dose group.Subcutaneous injection of estradiol benzoate was conducted to induce pseudo-estrus,and then 2×106 CFU·mL-1of Candida albicans was inoculated into vaginal lumen,followed by drug treatment for 7 days.Gram staining was used to observe the morphological changes of C.albicans in vagina;vaginal fungal load was detected on agar plate.Histological changes of vaginal tissues in mice were observed by HE staining.Lactate dehydrogenase(LDH),interleukin-6(IL-6)and tumor necrosis factor(TNF-α)levels in mouse lavage fluid were detected by enzyme-linked immunosorbent assay(ELISA).Neutrophils in vaginal lavage fluid was observed and counted by using Pap smear.The levels of IL-8 and MIP-2 in vaginal mucosa were detected by ELISA.IL-8 and MIP-2 mRNA levels in vaginal mucosa of mice were detected by qRT-PCR.The results showed that as compared with the normal group,VVC model group had a large number of hyphae and a high level of fungal loadinvagina.The vaginal mucosa was completely destroyed,the number of neutrophils increased,and the protein and mRNA levels of IL-8 and MIP-2 were up-regulated.After BAEB treatment,the hyphae of the treatment group was decreased,the fungal load was decreased,the impaired mucosa showed different degrees of improvement,the inflammatory factors were decreased to varying degrees,and the protein and mRNA levels of chemokine IL-8 and MIP-2 were down-regulated.In conclusion,BAEB may be used to treat VVC by inhibiting vulvovaginal candidiasis via blocking neutrophils recruitment into vagina.