目的 研究少腹逐瘀湯對(duì)輸卵管炎性不孕大鼠Nod樣受體蛋白3（NLRP3）炎癥小體的作用。方法 采用金黃色葡萄球菌法構(gòu)建大鼠輸卵管炎性不孕模型，造模成功雌性大鼠隨機(jī)分為3組：模型組、少腹逐瘀湯（以生藥計(jì)5g·kg^-1，臨床等效劑量）組、鹽酸左氧氟沙星（陽(yáng)性藥，80mg·kg^-1）組，另取10只大鼠作為對(duì)照組。各組于成模后第8天開始ig給藥，每天1次，連續(xù)30d，對(duì)照組和模型組ig等量0.9%氯化鈉注射液。各組雌鼠與成熟雄鼠按2∶1合籠，觀察受孕率；HE染色觀察各組大鼠輸卵管病理變化；Western blotting法檢測(cè)輸卵管組織中NLRP3蛋白表達(dá)；實(shí)時(shí)熒光定量PCR（qRTPCR）法檢測(cè)輸卵管組織中NLRP3和白細(xì)胞介素-1β（IL-1β）mRNA水平。原代培養(yǎng)大鼠輸卵管上皮細(xì)胞，10nmol·L^-1脂多糖（LPS）刺激24h制備炎癥模型，造模同時(shí)給予10mg·L^-1鹽酸左氧氟沙星、10mg·L^-1少腹逐瘀湯處理，Western blotting法檢測(cè)細(xì)胞NLRP3蛋白表達(dá)；qRT-PCR法檢測(cè)細(xì)胞中NLRP3、IL-1β、白細(xì)胞介素-6（IL-6）和腫瘤壞死因子-α（TNF-α）mRNA水平。結(jié)果 對(duì)照組的受孕率為60%，模型組的受孕率為40%，鹽酸左氧氟沙星組和少腹逐瘀湯組的受孕率均為80%。對(duì)照組大鼠輸卵管結(jié)構(gòu)清晰，黏膜皺褶豐富，黏膜上皮細(xì)胞排列整齊，管腔通暢；模型組大鼠的輸卵管結(jié)構(gòu)分界欠清，黏膜皺褶消失，黏膜上皮細(xì)胞排列紊亂，管腔狹窄，出現(xiàn)梗阻和大量炎性細(xì)胞浸潤(rùn)等情況；鹽酸左氧氟沙星組與少腹逐瘀湯組大鼠輸卵管結(jié)構(gòu)有改善，管壁組織結(jié)構(gòu)尚清晰，黏膜上皮細(xì)胞排列欠規(guī)整，管腔通暢，僅少量炎性細(xì)胞浸潤(rùn)。與對(duì)照組比較，模型組大鼠輸卵管組織的NLRP3蛋白和NLRP3、IL-1βmRNA水平顯著上調(diào)（P＜0.001）；與模型組比較，少腹逐瘀湯組輸卵管組織的NLRP3蛋白和NLRP3、IL-1βmRNA水平顯著下降（P＜0.05、0.001）。與對(duì)照組比較，模型組輸卵管上皮細(xì)胞NLRP3蛋白和NLRP3、IL-1β、IL-6、TNF-αmRNA水平顯著上調(diào)（P＜0.05、0.01、0.001）；與模型組比較，少腹逐瘀湯組NLRP3蛋白和NLRP3、IL-1β、IL-6、TNF-αmRNA水平顯著下降（P＜0.05、0.01）。結(jié)論 少腹逐瘀湯治療輸卵管炎性不孕癥的作用機(jī)制可能與調(diào)節(jié)NLRP3介導(dǎo)的炎癥反應(yīng)有關(guān)。

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

福建省自然科學(xué)基金青創(chuàng)項(xiàng)目(2020J05061);福建省中青年教師教育科研項(xiàng)目(JAT190249);福建省自然科學(xué)基金面上項(xiàng)目(2020J011041);正大天晴醫(yī)學(xué)項(xiàng)目研究(201511669);福建省衛(wèi)生計(jì)生科研人才培養(yǎng)項(xiàng)目-醫(yī)學(xué)創(chuàng)新課題(2018-CX-44)