摘要
目的本研究旨在观察补肾温阳化瘀方对大鼠肾阳虚血瘀型子宫内膜异位症(Endometriosis,EMs)模型的治疗作用,并以FoxO1介导的细胞自噬为切入点阐明其治疗EMs的可能作用机制。方法选择SPF级雌性SD大鼠100只,8-10周龄,随机分为假手术组、模型组、中药高剂量组、中药低剂量组和西药组,每组20只。采用自体移植法结合冰浴复制肾阳虚血瘀型EMs大鼠模型。造模成功后,中药高剂量组和中药低剂量组给予补肾温阳化瘀方颗粒水溶剂灌胃,西药组给予孕三烯酮悬浮液灌胃,假手术组和模型组给予同体积纯净水灌胃,各组连续灌胃3周。给药结束后采用机械性电子测痛仪测定机械痛阈值,计算脏器指数;HE染色观察在位内膜及异位灶的病理形态学改变;免疫组化检测在位内膜及异位灶组织FoxO1、NF-κB、Beclin-1、LC3的蛋白定位表达;RT-PCR检测在位内膜及异位灶FoxO1、NF-κB、Beclin-1、LC3-I及LC3-II mRNA的表达水平;Western Blot测定在位内膜及异位病灶FoxO1、NF-κB、Beclin-1、LC3-I及LC3-II蛋白的表达水平。结果与假手术组相比,模型组FoxO1、NF-κB、Beclin-1、LC3-I及LC3-II蛋白表达及mRNA水平显著升高,差异具有统计学意义(P<0.05)。与模型组相比,中药高剂量组、中药低剂量组在位内膜及异位灶的FoxO1、NF-κB、Beclin-1、LC3-I及LC3-II蛋白表达及mRNA水平显著下降,差异具有统计学意义(P<0.05)。结论FoxO1、NF-κB及自噬相关因子Beclin-1、LC3参与EMs疾病的的发展并具有协同作用,补肾温阳化瘀方具有改善EMs的作用,这种作用可能与降低FoxO1、NF-κB的表达从而抑制细胞自噬水平有关。
Objective We aimed to evaluate the therapeutic effect of Bushen Wenyang Huayu Recipe(BWHR)on a rat model of endometriosis(EM)that involves kidney yang deficiency and blood stasis and determine whether its mechanism of action involves FoxO1-mediated autophagy.Methods Female-specific pathogen-free SD rats aged 8-10 weeks were randomly allocated to a sham surgery group,a model group,a high-dose BWHR group,a low-dose BWHR group,or a western medicine group(n=20).A rat model of EM associated with kidney yang deficiency and blood stasis was established by autologous transplantation combined with an ice bath.After confirmation of the successful induction of EM,the high-dose and low-dose groups were administered BWHR,the western medicine group was administered gestrinone suspension,and the sham and model groups were administered the same volume of water by gavage for 3 weeks.Subsequently,the mechanical pain threshold of the rats was measured using a mechanical electronic pain meter,and organ indices were calculated.The histopathological changes in the eutopic endometrium and ectopic foci were evaluated on hematoxylin and eosin-stained sections.The expression of FoxO1,NF-κB,Beclin-1 and LC3 in eutopic endometrium and ectopic lesions was assessed using immunohistochemistry.The mRNA and protein expression of FoxO1,NF-κB,Beclin-1,LC3-I,and LC3-II was measured using Real time PCR and Western Blot,respectively.Results The protein and mRNA expression of FoxO1,NF-κB,Beclin-1,LC3-I,and LC3-II was significantly higher in the model group than in the control group(P<0.05).However,the protein and mRNA expression of FoxO1,NF-κB,Beclin-1 and LC3 in eutopic endometrium and ectopic lesions in the high-dose and low-dose BWHR groups were significantly lower(P<0.05).Conclusions FoxO1,NF-κB,and the autophagy related factors Beclin-1 and LC3 may be involved in the development of EMs and have synergistic effects.BWHR may ameliorate EM by reducing the expression of FoxO1 and NF-κB,which inhibits autophagy.
作者
孟鑫
李颖
张宇
曹璨
白羽
陈景伟
MENG Xin;LI Ying;ZHANG Yu;CAO Can;BAI Yu;CHEN Jingwei(Hebei Key Laboratory of Integrative Medicine on Liver-Kidney Patterns,Institute of Integrative Medicine,Hebei University of Chinese Medicine,Shijiazhuang 050200,China)
出处
《中国实验动物学报》
CAS
CSCD
北大核心
2021年第4期417-427,共11页
Acta Laboratorium Animalis Scientia Sinica
基金
国家自然科学基金项目(82074483)
河北省重点研发计划项目卫生健康创新专项(21377725D)
省级大学生创新创业训练计划项目(202014432067)。
