摘要
目的基于辅助性T细胞17(Th17)/调节性T细胞(Treg)失衡探讨补肾固胎合剂治疗复发性流产(RSA)的可能作用机制。方法将80只SD大鼠(30只雄鼠、50只雌鼠)适应性喂养1周后进行合笼,检测成功怀孕后记为妊娠第0天,并随机分为空白对照组、模型组、地屈孕酮组、补肾固胎合剂低剂量组以及补肾固胎合剂高剂量组共5组,每组各10只。妊娠第1天至第9天下午,除空白对照组外,其他组均灌胃剂量为450 mg·kg^(-1)·d^(-1)的羟基脲片水溶液,妊娠第10天上午均灌胃4 mg·kg^(-1)·d^(-1)米非司酮水溶液。妊娠第1天至第14天,上午空白对照组与模型组均灌胃等体积生理盐水,地屈孕酮组灌胃3.02 mg·kg^(-1)·d^(-1)地屈孕酮水溶液,补肾固胎合剂低剂量组和高剂量组分别给予9.9 g·kg^(-1)·d^(-1),39.6 g·kg^(-1)·d^(-1)补肾固胎合剂水煎剂灌胃。观察孕鼠妊娠14天的体质量变化;计算孕鼠脾脏指数;观察各组孕鼠胚胎生长发育状况;对孕鼠蜕膜组织和脾组织进行苏木素-伊红(HE)染色,并观察其病理学变化;ELISA试剂盒检测孕鼠血清中白介素-17(IL-17)、肿瘤坏死因子-α(TNF-α)、白介素-10(IL-10)、转化生长因子-β1(TGF-β1)含量;免疫荧光染色(IF)检测蜕膜组织中视黄酸相关孤儿核受体γt(RORγt)、叉状头螺旋转录因子3(FOXP3)蛋白表达情况;蛋白免疫印迹法(Western blot)检测孕鼠蜕膜组织中IL-17、TNF-α、IL-10、TGF-β1蛋白表达。结果与空白对照组比较,模型组孕鼠第14天体质量极显著降低(P<0.01);脾脏指数极显著降低(P<0.01);孕鼠胚胎发育大小不均匀,部分胚胎出现瘀血、坏死情况;胚胎丢失率极显著升高(P<0.01);蜕膜组织细胞排列杂乱,胞核固缩,部分消失,胞质水肿,部分呈空泡样变;脾组织结构严重异常,部分脾小结结构紊乱,红髓与白髓界限杂乱、模糊不清;血清、蜕膜组织中IL-17、TNF-α、RORγt表达量,IL-17/TGF-β1、TNF-α/IL-10、RORγt/FOXP3比�
Objective Based on the imbalance of helper T cell 17(Th17)/regulatory T cell(Treg),the possible mechanism of Bushen Gutai Mixture in the treatment of recurrent abortion(RSA)was investigated.Methods After 1 week of adaptive feeding,80 SD rats(30 male and 50 female)were combined with cages,and the mark after successful pregnancy was detected as day 0 of gestation,and were randomly divided into 5 groups with 10 rats in each group,including blank control group,model group,dydrogesterone group,low-dose Bushen Gutai mixture group and high-dose Bushen Gutai mixture group.In the afternoon of the 1st to 9th day of gestation,except the blank control group,the other groups were intragaxed with 450 mg·kg^(-1)·d^(-1) hydroxyurea tablet aqueous solution,and intragaxed with 4 mg·kg^(-1)·d^(-1) mifepristone aqueous solution in the morning of the 10th day of gestation.From day 1 to day 14 of gestation,the blank control group and model group were given equal volume normal saline in the morning,the dydrogesterone group was given 3.02 mg·kg^(-1)·d^(-1) dydrogesterone aqueous solution,and the low dose group and high dose group were given 9.9 g·kg^(-1)·d^(-1) and 39.6 g·kg^(-1)·d^(-1) water decoction of Bushen Gutai mixture,respectively.The changes of body mass at 14 days gestation were observed.The spleen index of pregnant mice was calculated.The embryonic growth and development of pregnant mice in each group were observed.He⁃matoxylin-eosin(HE)staining was performed on decidua tissue and spleen tissue of pregnant mice,and the pathological changes were observed.The contents of interleukin-17(IL-17),tumor necrosis factor-α(TNF-α),interleukin-10(IL-10)and transfor⁃ming growth factor-β1(TGF-β1)in serum of pregnant mice were detected by ELISA kit.The expression of retinoid-related orphan nuclear receptorγt(RORγt)and forked head spiral transcription factor 3(FOXP3)in decidual tissue was detected by immunofluores⁃cence staining(IF).The expressions of IL-17,TNF-α,IL-10 and TGF-β1 in decidua of pregnant mice were dete
作者
潘琴
萧闵
江晓翠
姜兴宇
甘雨娟
姚国晋
田文静
张玮珩
张迎春
PAN Qin;XIAO Min;JIANG Xiao-cui;JIANG Xing-yu;GAN Yu-juan;YAO Guo-Jin;TIAN Wen-jing;ZHANG Wei-heng;ZHANG Ying-chun(First Clinical College,Hubei University of Chinese Medicine,Wuhan 430065,China;Experimental Centre of Traditional Chinese Medicine,Hubei University of Chinese Medicine,Wuhan 430065,China;Hubei Shizhen Laboratory,Hubei University of Chinese Medicine,Wuhan 430065,China;Hubei Maternal and Child Health Care Hospital,Wuhan 430070,China;School of Basic Medicine,Hubei University of Chinese Medicine,Wuhan 430065,China)
出处
《时珍国医国药》
CAS
CSCD
北大核心
2024年第13期2946-2954,共9页
Lishizhen Medicine and Materia Medica Research
基金
2022年全国名老中医药专家传承工作室建设项目(国中医药人教函[2022]75号)
湖北中医药大学中医药传承创新计划思政专项(2022SZXC001)。
