摘要
目的:研究补肾中药复方右归丸对自然衰老致卵巢功能低下的大鼠血管生成的影响与CXC趋化因子8(CXCL8)/趋化因子受体1/2(CXCR1/2)信号通路及血管生成素-1(Ang-1),血管生成素-2(Ang-2)的关系,探讨其改善卵巢功能的作用机制。方法:将56只雌性SD大鼠随机分为青年组8只、自然衰老致卵巢功能低下的雌性大鼠模型组48只,青年组大鼠常规饲养,48只模型组大鼠常规饲养过程中做5~7 d阴道脱落细胞学涂片,将连续4个动情周期阴道细胞学表现为动情周期长、之后持续动情、反复假妊娠的大鼠作为初老大鼠,连续10 d表现为角化细胞指数高于50%的青年大鼠作为青年组。模型建立后,将存活造模大鼠随机分为初老空白组,结合雌激素组(65μg·kg^(-1)·d^(-1)),左归丸组(33 g·kg^(-1)·d^(-1)),右归丸低、中、高剂量组(1.2,2.4,4.8 g·kg^(-1)d^(-1)),青年组和初老空白组以等体积生理盐水灌胃,持续30 d。实验结束后,苏木素-伊红(HE)染色法观察大鼠卵巢组织形态学变化;蛋白免疫印迹法(Western blot)检测大鼠卵巢组织中趋化因子CXCL8,CXCR1,CXCR2,Ang-1,Ang-2的蛋白表达水平;免疫组化标记大鼠卵巢组织中趋化因子CXCL8,CXCR1,CXCR2,Ang-1,Ang-2蛋白的表达;实时荧光定量聚合酶链式反应(Real-time PCR)检测大鼠卵巢组织中趋化因子CXCL8,CXCR1,CXCR2,Ang-1,Ang-2 mRNA表达。结果:与青年组比较,初老空白组各级生长卵泡数、黄体数、血管数目数量少且闭锁卵泡多(P<0.01),卵巢组织CXCL8,CXCR1及CXCR2水平蛋白和mRNA水平显著上升(P<0.01),卵巢组织中Ang-1及Ang-2蛋白和mRNA水平明显下降(P<0.05);与初老空白组相比,各用药组初老大鼠的各级生长卵泡数、黄体数、血管数目明显增加(P<0.05),而闭锁卵泡数明显减少(P<0.05),卵巢组织CXCL8,CXCR1及CXCR2水平蛋白和mRNA水平明显下降(P<0.05),卵巢组织中Ang-1及Ang-2蛋白和mRNA水平明显上升(P<0.05)。结论:右归丸可以通过下�
Objective:To study the effects of Chinese herbal compound Youguiwan on angiogenesis of rats with ovarian dysfunction caused by natural aging and its relationship with chemokine interleukin 8(CXCL8)/CXC chemokine receptor 1/2(CXCR1/2) signaling pathway, angiopoietin 1(Ang-1), and angiopoietin 2(Ang-2),so as to explore its mechanism in improving the ovarian function. Method: Fifty six female SD rats were randomly divided into the young control group(n=8)and modeling group(n=48,ovarian dysfunction caused by natural aging). Rats in both the young control and modeling groups were routinely fed,during which the ones in the modeling group underwent exfoliative cytology of vaginal smears for five to seven days. The ones presented with prolonged estrous cycle, followed by continuous estrus and repeated pseudopregnancy revealed by vaginal cytology during four consecutive estrous cycles indicated early aging,and the young rats with keratinocyte proliferation index higher than 50% for 10 consecutive days were classified into the young control group. The successfully modeled rats were randomly divided into the early-aged group,estrogen(65 μg·kg^(-1)·d^(-1))group,Zuoguiwan(33 g·kg^(-1)·d^(-1))group,as well as the low-,medium-,and highdose(1.2,2.4,4.8 g·kg^(-1)·d^(-1))Youguiwan groups. Rats in the young control group and the early-aged group were gavaged with the same volume of normal saline for 30 days. After the experiment,the morphological changes in rat ovary were observed by hematoxylin-eosin(HE)staining. The protein expression levels of chemokines CXCL8,CXCR1,CXCR2,Ang-1,and Ang-2 in rat ovary were detected by Western blotting and immunohistochemistry,and the mRNA expression levels of CXCL8,CXCR1,CXCR2,Ang-1,and Ang-2 by real-time fluorescence quantitative polymerase chain reaction(Real-time PCR). Result: Compared with the young control group,the early-aged group exhibited reduced number of growing follicles,corpus luteum,and blood vessels at all levels,elevated atretic follicles(P<0.01),up-regulated protein and mR
作者
魏小晶
段恒
唐立明
宋谦谦
WEI Xiao-jing;DUAN Heng;TANG Li-ming;SONG Qian-qian(College of Traditional Chinese Medicine(TCM),Chongqing Key Laboratory of TCM for Prevention and Cure of Metabolic Diseases,Chongqing 400016,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2022年第1期50-57,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
2020年重庆市研究生科研创新项目(CYS20216)
重庆市基础科学与前沿技术研究一般项目(cstc2017jcyjAX0350)。
