补肾活血方对不明原因复发性流产小鼠蜕膜化过程中p53、p21的影响  

Effects of Bushen Huoxue Decoction on p53 and p21 in decidualization of mice with unexplained recurrent abortion

作　　者：常卓 朱慧[1] 周雪明[1] 张杨[2] 王炜 陈璐[2] 姜北[1] 冯晓玲[3] CHANG Zhuo;ZHU Hui;ZHOU Xue-ming;ZHANG Yang;WANG Wei;CHEN Lu;JIANG Bei;FENG Xiao-ling(Heilongjiang University of Chinese Medicine,Harbin 151000,China;First Affiliated Hospital,Heilongjiang University of Chinese Medicine,Harbin 151000,China)

机构地区：[1]黑龙江中医药大学,哈尔滨151000 [2]黑龙江中医药大学附属第一医院,哈尔滨151000 [3]黑龙江中医药大学附属第一医院妇科二科,哈尔滨151000

出　　处：《中华中医药杂志》2022年第1期473-477,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81973894,No.81803999);黑龙江中医药大学校科研基金资助项目(No.2019MS34)。

摘　　要：目的:观察在不明原因复发性流产(URSA)小鼠蜕膜化过程中补肾活血方对p53、p21的影响,探讨补肾活血方治疗URSA的作用机制。方法:将动情周期稳定的CBA/J雌鼠分别与BALB/c、DBA/2雄鼠合笼,构建正常妊娠模型与URSA妊娠模型,正常妊娠雌鼠记做对照组(25只),URSA妊娠雌鼠随机分为模型组(25只)、中药组(25只)。3组雌鼠于发现妊娠第1~8天进行灌胃(对照组、模型组予0.9%氯化钠溶液,中药组予补肾活血方中药水煎剂)。于蜕膜化全过程(妊娠第4~8天)连续每天每组处死5只小鼠并取材。免疫组化分析妊娠第8天蜕膜中p53、p21的阳性表达,RT-PCR、Western Blot分别检测妊娠第4~8天小鼠蜕膜组织中p53、p21 mRNA及蛋白变化。结果:妊娠第8天,对照组、中药组p53、p21蛋白阳性面积百分率显著低于模型组(P<0.05);妊娠第6~8天,对照组、中药组p53、p21 mRNA水平与蛋白表达均显著低于模型组(P<0.05)。结论:补肾活血方体内干预可参与调节URSA小鼠的蜕膜化过程,可显著降低蜕膜化中后期的p53、p21表达。Objective:To observe the effects of Bushen Huoxue Decoction on p53,p21 in unexplained recurrent spontaneous abortion(URSA)in process of mice decidualization and to discussion on the mechanism of Bushen Huoxue Decoction in treating URSA diseases.Methods:The CBA/J female mice with stable emotional cycle were caged with BALB/c and DBA/2 males to build the normal pregnancy female mice model and URSA pregnant female mice model.Normal pregnant female mice were recorded as control group(25 mice),whereas URSA pregnant females were randomly divided into model group(25 mice)and traditional Chinese medicine group(25 mice).Three groups of female mice were perfused from d1 to d8 of pregnancy(control group,model group were perfused with normal saline,and traditional Chinese medicine group was perfused with Bushen Huoxue Decoction).In the whole process of decidualization(from d4 to d8 of pregnancy),each group was executed with 5 mice for drawing materials every day.With immunohistochemical analysis of positive expression of p53 and p21 in d8 during pregnancy,by adopting methods of RT-PCR and Western Blot,the mRNA and protein changes of p53,p21 in decidual tissue of mice on d4 to d8 day of pregnancy were detected.Results:On the 8 th day of pregnancy,the percentage of positive area of p53 and p21 protein in control group and traditional Chinese medicine group was significantly lower than that in model group(P<0.05);On the d6 to d8 day of pregnancy,p53,p21 mRNA and protein expression of control group and traditional Chinese medicine group were lower than that in the model group(P<0.05).Conclusion:The in vivo intervention of Bushen Huoxue Decoction can regulate the decidualization process of URSA mice and significantly reduce the p53,p21 expression in the middle and late stages of decidualization.

关 键 词：不明原因复发性流产 蜕膜化 补肾活血方 P53 P21 机制 动物模型 

分 类 号：R285.5[医药卫生—中药学]