目的 研究少腹逐瘀汤对输卵管炎性不孕大鼠Nod样受体蛋白3（NLRP3）炎症小体的作用。方法 采用金黄色葡萄球菌法构建大鼠输卵管炎性不孕模型，造模成功雌性大鼠随机分为3组：模型组、少腹逐瘀汤（以生药计5g·kg^-1，临床等效剂量）组、盐酸左氧氟沙星（阳性药，80mg·kg^-1）组，另取10只大鼠作为对照组。各组于成模后第8天开始ig给药，每天1次，连续30d，对照组和模型组ig等量0.9%氯化钠注射液。各组雌鼠与成熟雄鼠按2∶1合笼，观察受孕率；HE染色观察各组大鼠输卵管病理变化；Western blotting法检测输卵管组织中NLRP3蛋白表达；实时荧光定量PCR（qRTPCR）法检测输卵管组织中NLRP3和白细胞介素-1β（IL-1β）mRNA水平。原代培养大鼠输卵管上皮细胞，10nmol·L^-1脂多糖（LPS）刺激24h制备炎症模型，造模同时给予10mg·L^-1盐酸左氧氟沙星、10mg·L^-1少腹逐瘀汤处理，Western blotting法检测细胞NLRP3蛋白表达；qRT-PCR法检测细胞中NLRP3、IL-1β、白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）mRNA水平。结果 对照组的受孕率为60%，模型组的受孕率为40%，盐酸左氧氟沙星组和少腹逐瘀汤组的受孕率均为80%。对照组大鼠输卵管结构清晰，黏膜皱褶丰富，黏膜上皮细胞排列整齐，管腔通畅；模型组大鼠的输卵管结构分界欠清，黏膜皱褶消失，黏膜上皮细胞排列紊乱，管腔狭窄，出现梗阻和大量炎性细胞浸润等情况；盐酸左氧氟沙星组与少腹逐瘀汤组大鼠输卵管结构有改善，管壁组织结构尚清晰，黏膜上皮细胞排列欠规整，管腔通畅，仅少量炎性细胞浸润。与对照组比较，模型组大鼠输卵管组织的NLRP3蛋白和NLRP3、IL-1βmRNA水平显著上调（P＜0.001）；与模型组比较，少腹逐瘀汤组输卵管组织的NLRP3蛋白和NLRP3、IL-1βmRNA水平显著下降（P＜0.05、0.001）。与对照组比较，模型组输卵管上皮细胞NLRP3蛋白和NLRP3、IL-1β、IL-6、TNF-αmRNA水平显著上调（P＜0.05、0.01、0.001）；与模型组比较，少腹逐瘀汤组NLRP3蛋白和NLRP3、IL-1β、IL-6、TNF-αmRNA水平显著下降（P＜0.05、0.01）。结论 少腹逐瘀汤治疗输卵管炎性不孕症的作用机制可能与调节NLRP3介导的炎症反应有关。

Objective To study the effect of Shaofu Zhuyu Decoction on NOD-like receptor protein 3 (NLRP3) inflammasome in salpingitis infertility rats.Methods Staphylococcus aureus method was used to construct a rat tubal inflammatory infertility model, and the female model rats were randomly divided into three groups: model group, Shaofu Zhuyu Decoction (5 g·kg^-1 crude drug, clinical equivalent dose) group, and levofloxacin hydrochloride (80 mg·kg^-1, positive drug) group, and 10 normal rats were selected as control group. Each group was given ig on the 8th day after modeling, once a day, for 30 days, control group and model group were given the same amount of 0.9% sodium chloride injection. After 30 days of continuous administration, female mice and mature male mice in each group were combined in cage at 2∶ 1 to observe the conception rate. HE staining was used to observe the pathological changes of oviduct in each group. NLRP3 protein expression in fallopian tubes was detected by Western blotting. Realtime quantitative fluorescence PCR (qRT-PCR) was used to detect NLRP3 and interleukin-1β (IL-1β) mRNA levels in fallopian tubes. The primary cultured rat oviduct epithelial cells were stimulated with 10 nmol·L^-1 lipopolysaccharide (LPS) for 24 h to make an inflammatory model. At the same time, the cells were treated with 10 mg·L^-1 levofloxacin hydrochloride group and 10 mg·L^-1 Shaofu Zhuyu Decoction. The NLRP3 protein expression was detected by Western blotting method. The mRNA levels of NLRP3, IL-1β, interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected by qRT-PCR.Results The pregnancy rate of the normal control group was 60%, that of the model group, Shaofu Zhuyu Decoction group, and levofloxacin hydrochloride group was 40%, 80%, 80% respectively. In control group, the structure of fallopian tube was clear, the mucosal folds were abundant, the mucosal epithelial cells were arranged neatly, and the lumen was unobstructed. In the model group, the structure boundary of fallopian tube was not clear, the mucosal folds disappeared, the arrangement of mucosal epithelial cells was disordered, the lumen was narrow, obstruction and a large number of inflammatory cell infiltration occurred. In Shaofu Zhuyu Decoction and levofloxacin hydrochloride group group, the structure of fallopian tube was improved, the tissue structure of tube wall was clear, the arrangement of mucosal epithelial cells was irregular, the lumen was unobstructed, and only a small amount of inflammatory cells infiltrated. Compared with control group, NLRP3 protein expression, NLRP3 and IL-1β mRNA level in model group were significantly upregulated (P <0.001). Compared with model group, NLRP3 protein expression, NLRP3 and IL-1β mRNA level in tubal tissue of Shaofu Zhuyu Decoction group were significantly decreased (P <0.05, 0.001). Compared with control group, NLRP3 protein expression and mRNA levels of NLRP3, IL-1β, IL-6 and TNF-α were significantly up-regulated in model group (P <0.05, 0.01, 0.001). Compared with model group, NLRP3 protein expression and mRNA levels of NLRP3, IL-1β, IL-6 and TNF-α in Shaofu Zhuyu Decoction group were significantly decreased (P <0.05, 0.01).Conclusion The mechanism of Shaofu Zhuyu Decoction in the treatment of salpingitc infertility may be related to the regulation of NLRP3 mediated inflammatory response.

R285.5

福建省自然科学基金青创项目(2020J05061);福建省中青年教师教育科研项目(JAT190249);福建省自然科学基金面上项目(2020J011041);正大天晴医学项目研究(201511669);福建省卫生计生科研人才培养项目-医学创新课题(2018-CX-44)