左归丸促初老大鼠生殖器官血管生成与EPCs促动员因素GM-CSF、SDF-1及其受体关系的研究  

Relationship between promoting effect of Zuogui Pills on angiogenesis of reproductive organs and mobilization factors GM-CSF,SDF-1,and their receptors of EPCs in early-aging rats

作　　者：宋谦谦 段恒[1] 彭万春 魏小晶 SONG Qian-qian;DUAN Heng;PENG Wan-chun;WEI Xiao-jing(Chongqing Key Laboratory of Traditional Chinese Medicine Prevention and Treatment of Metabolic Diseases,College of Traditional Chinese Medicine,Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学中医药学院中医药防治代谢性疾病重庆市重点实验室,重庆400016

出　　处：《中国中药杂志》2023年第16期4467-4474,共8页China Journal of Chinese Materia Medica

基　　金：重庆市科卫联合项目(2021ZY023852)。

摘　　要：该实验旨在研究左归丸促进初老大鼠生殖器官卵巢、阴道血管生成与内皮祖细胞(EPCs)促动员因素粒-巨噬细胞集落刺激因子(GM-CSF)、基质细胞衍生因子-1(SDF-1)及其受体的关系,探讨左归丸改善初老大鼠生殖功能低下的作用机制。采用超高效液相色谱-串联质谱(UHPLC-MS/MS)对左归丸浸膏进行中药化学成分分析。然后选用14月龄动情周期紊乱的雌性初老大鼠40只,随机分为初老空白组、结合雌激素组、左归丸低剂量组、左归丸高剂量组,每组各10只;另外选用10只4月龄雌性大鼠作为青年对照组。初老空白组和青年对照组均用20 g·kg-1蒸馏水灌胃,结合雌激素组用65μg·kg-1结合雌激素悬液进行灌胃,低剂量组用11g·kg-1左归丸浸膏灌胃,高剂量组用33 g·kg-1左归丸浸膏进行灌胃,每日1次,连续灌胃15 d。收集各组大鼠血清,用酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)法检测血清中促EPCs动员因素SDF-1、GM-CSF水平;苏木精-伊红(hematoxylin-eosin,HE)染色观察卵巢各级卵泡数目、黄体数目、阴道上皮层数、阴道皱襞数以及卵巢、阴道组织血管等在用药组的变化;蛋白免疫印迹(Western blot)法检测卵巢、阴道组织雌激素受体(ER)、粒-巨噬细胞集落刺激因子受体(GM-CSFR)、趋化因子受体(CXCR)4、CXCR7蛋白表达水平。结果发现,初老空白组大鼠黄体数、各级生长卵泡数和血管数减少(P<0.05);阴道黏膜厚度、上皮层数、皱襞数、固有层血管数明显减少(P<0.05);外周血中SDF-1、GM-CSF含量明显降低(P<0.05);大鼠卵巢、阴道组织中ER、CXCR4、CXCR7、GM-CSFR蛋白水平明显下降(P<0.05)。各用药组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),阴道黏膜厚度、上皮层数、阴道黏膜皱襞数和固有层血管数增加(P<0.05);外周血中SDF-1、GM-CSF含量升高(P<0.05);卵巢、阴道组织中ER、This study aimed to investigate the relationship between the promoting effect of Zuogui Pills on ovarian and vaginal angiogenesis in early-aging rats and mobilization factors granulocyte-macrophage colony-stimulating factor(GM-CSF),stromal cell-derived factor-1(SDF-1),and their receptors of endothelial progenitor cells(EPCs)and explore the mechanism of Zuogui Pills in improving reproductive hypofunction in early-aging rats.Ultra-high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS)was used to analyze the chemical components of the extract of Zuogui Pills.Forty 14-month-old female early-aging rats with estrous cycle disorder were randomly divided into a blank group,a conjugated estrogen group(conjugated estrogen suspension,65μg·kg-1),and low-(11 g·kg-1)and high-dose(33 g·kg-1)Zuogui Pills groups,with 10 rats in each group.In addition,104-monthold female rats were assigned to the youth control group.The rats in the blank group and the youth control group were treated with 20 g·kg-1 distilled water by gavage,while those in the groups with drug intervention were treated with corresponding drugs by gavage,once a day for 15 days.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of SDF-1 and GM-CSF in the mobilization of EPCs in serum.Hematoxylin-eosin(HE)staining was used to observe the changes in the number of ovarian follicles at all levels and corpus luteum,the number of vaginal epithelial layers,the number of vaginal folds,and the blood vessels of ovarian and vaginal tissues in the groups with drug intervention.Western blot was used to detect the expression of ER,GM-CSFR,CXCR4,and CXCR7 proteins in ovarian and vaginal tissues.As revealed by the results,the blank group showed decreased number of corpus luteum,growing follicles at all levels,and blood vessels(P<0.05),decreased thickness of vaginal mucosa,the number of epithelial layers,the number of vaginal folds,and the number of vessels in the lamina propria(P<0.05),reduced content of SDF-1 and GM-CSF in the periphera

关 键 词：左归丸 生殖器官血管生成 内皮祖细胞 粒-巨噬细胞集落刺激因子 基质细胞衍生因子-1 

分 类 号：R285.5[医药卫生—中药学]