瓜蒌牛蒡汤对哺乳期乳腺炎小鼠的改善作用及机制研究  

Study on the Improvement Effect and It’s Mechanism of Gualou Niubang Decoction on Lactation Mastitis Mice

作　　者：张立清 李婕 洪秀宇 聂娟 肖建琴 ZHANG Liqing;LI Jie;HONG Xiuyu(Department of Surgical,Anji County Chinese Medicine Hospital,Anji County,Zhejiang Province 313300;不详)

机构地区：[1]浙江省安吉县中医医院外科,313300 [2]浙江省安吉县中医医院医教科,313300

出　　处：《医学理论与实践》2023年第1期1-5,9,共6页The Journal of Medical Theory and Practice

基　　金：湖州市公益性应用研究项目(2019GY51)。

摘　　要：目的:通过体内外实验探讨瓜蒌牛蒡汤治疗哺乳期乳腺炎(LA)的作用机制。方法:制备瓜蒌牛蒡汤和头孢曲松钠含药血清,脂多糖(LPS)诱导小鼠乳腺上皮细胞EMT6细胞模拟体外乳腺炎细胞模型后,用5%、10%、15%瓜蒌牛蒡汤血清和10%头孢曲松钠血清干预24h,CCK8实验测定细胞活性,ELISA测定细胞上清液白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)、IL-1β含量。另将LPS注入产后5~7d的ICR雌鼠乳腺进行LA造模,随后瓜蒌牛蒡汤和头孢曲松钠灌胃治疗4周,行HE染色观察雌鼠乳腺组织炎症病理变化,ELISA测定血清IL-12、IL-10、IL-1β含量。Western blot测定细胞和组织中Toll样受体4(TLR4)、核因子κB(NF-κB-p65)、p-NF-κB-p65、TNF-α蛋白表达。结果:不同剂量瓜蒌牛蒡汤或含药血清干预下,乳腺炎细胞活力显著上升(P<0.05或P<0.01),IL-6、TNF-α、IL-1β含量显著下降(P<0.05或P<0.01),乳腺炎组织病理损伤减轻,血清IL-12、IL-1β含量显著下降(P<0.05或P<0.01),IL-10含量显著上升(P<0.05或P<0.01);乳腺炎细胞和组织中TLR4、p-NF-κB-p65/NF-κB-p65、TNF-α蛋白表达显著下降(P<0.05或P<0.01)。结论:瓜蒌牛蒡汤对LA的改善作用可能与抑制TLR4/NF-κB信号通路有关。Objective:To investigated and explore the effective and mechanism of Gualou Niubang Decoction(GND)on lactation mastitis in vitro and vivo.Methods:Preparation of containing GND or ceftriaxone sodium serum respectively.Mouse mammary epithelial EMT6 cells were induced by lipopolysaccharides(LPS)to model mastitis cell,then intervention with containing 5%,10%,15%GND and 10%ceftriaxone sodium serum for 24 hours,CCK8 test was used to detect cell viability,ELISA was used to detect IL-6,TNF-αand IL-1βlevels.In addition,LPS was injected into the mammary glands of ICR female mice at 5~7 days postpartum to establish LA model mice,then mice were gavaged with 7.5,15,30g/kg GND and ceftriaxone sodium(100mg/kg)for 4 weeks,HE staining was used to observe pathological changes of the mammary glands,and ELISA was used to detect serum IL-12,IL-10 and IL-1βlevels.Western blot was used to determine the protein expressions of TLR4,NF-κB-p65,p-NF-κB-p65 and TNF-αin mastitis cells and tissue.Results:Intervention with different dose of GND and containing ceftriaxone sodium serum,the viability of mastitis cells significantly increased(P<0.05 or P<0.01),and IL-6,TNF-αand IL-1βlevels significantly decreased(P<0.05 or P<0.01),mastitis tissue also LA mice serum IL-12 and IL-1βlevels significantly decreased(P<0.05 or P<0.01),while IL-10 level significantly increased(P<0.05 or P<0.01);in addition,protein expression of TLR4,p-NF-κB-p65/NF-κB-p65 and TNF-αin mastitis cells and tissues were significantly decreased(P<0.05 or P<0.01).Conclusion:The improvement of Gualou Niubang Decoction(GND)on LA may related to inhibition of TLR4/NF-κB pathway.

关 键 词：哺乳期乳腺炎 瓜蒌牛蒡汤 TLR4/NF-κB信号通路 炎症因子 

分 类 号：R-33[医药卫生] R711