摘要
目的基于肿瘤转移抑制基因Kisspeptin/G蛋白偶联受体54(GPR54)系统探究当归有效组分对原发性痛经(PD)模型小鼠下丘脑-垂体-卵巢轴(HPOA)的调控作用。方法采用戊酸雌二醇联合缩宫素建立小鼠原发性痛经模型,将72只雌性未孕KM小鼠随机分为正常组、模型组、阳性对照组(67 mg·kg^(-1)布洛芬灌胃给药)和低、中、高剂量实验组(分别用150、300、600 mg·kg^(-1)当归效应组分灌胃给药),每组12只。以酶联免疫吸附法测定各组小鼠血清中雌二醇(E2)、孕酮(P)的含量;以免疫组化法检测各组小鼠下丘脑GPR54蛋白表达水平;以逆转录聚合酶链反应法检测各组小鼠下丘脑组织中GPR54 mRNA水平的表达。结果正常组、模型组、阳性对照组和低、中、高剂量实验组血清中E2含量分别为(23.62±2.26)、(30.29±2.57)、(24.49±2.13)、(29.46±3.68)、(27.04±3.57)和(27.15±3.25)pmol·L^(-1);P含量分别为(885.13±78.71)、(522.63±61.33)、(716.96±56.80)、(588.81±81.11)、(591.63±43.69)和(634.30±84.96)pmol·L^(-1);下丘脑GPR54蛋白表达水平分别为0.17±0.03、0.32±0.04、0.25±0.04、0.26±0.04、0.25±0.05和0.21±0.03;GPR54蛋白表达水平分别为1.00±0.03、1.46±0.12、1.28±0.09、1.41±0.09、1.36±0.07和1.35±0.08。模型组上述指标与正常组比较,差异均有统计学意义(均P<0.01);中、高剂量实验组上述指标与模型组比较,差异均有统计学意义(P<0.01,P<0.05)。基因结果的趋势与蛋白一致。结论当归有效组分可有效防治PD,其机制可能与下调下丘脑-垂体-卵巢轴Kisspeptin/GPR54系统表达有关。
Objective To investigate the regulatory effect of Angelica sinensis effective components on hypothalamic-pituitary-ovarian axis in primary dysmenorrhea model mice based on kisspeptin/GPR54 system.Methods The mouse model of primary dysmenorrhea was established by estradiol valerate combined with oxytocin.And then the 72 unfertilized female KM mice were randomly divided into the normal group,the model group,positive control group(67 mg·kg^(-1)ibuprofen irrigation stomach),and experimental-L,-M,-H groups(150,300,600 mg·kg^(-1)Angelica sinensis effective components irrigation stomach),with 12 mice in each group.Enzyme-linked immunosorbent assay was used to measure the levels of E2,P in serum of mice.Immunohistochemical method was used to detect the expression levels of GPR54 in hypothalamus.Reverse transcription polymerase chain reaction was also used to detect the expression of GPR54 mRNA in hypothalamus tissues.Results The serum E2content in normal group,model group,positive control group and experimental-L,-M,-H groups were(23.62±2.26),(30.29±2.57),(24.49±2.13),(29.46±3.68),(27.04±3.57)and(27.15±3.25)pmol·L^(-1),respectively;P content were(885.13±78.71),(522.63±61.33),(716.96±56.80),(588.81±81.11),(591.63±43.69)and(634.30±84.96)pmol·L^(-1),respectively;the relative expression of GPR54 protein in hypothalamus tissues were 0.17±0.03,0.32±0.04,0.25±0.04,0.26±0.04,0.25±0.05 and 0.21±0.03,respectively;the relative expression of GPR54 mRNA in hypothalamus tissues were 1.00±0.03,1.46±0.12,1.28±0.09,1.41±0.09,1.36±0.07 and 1.35±0.08,respectively.Compared with the control group,the above indexes in the model group were all significantly different(all P<0.01);Compared with the model group,the above indexes in the experimental-M,-H groups were all significantly different(P<0.01,P<0.05).The trend of gene results is consistent with that of protein.Conclusion The Angelica sinensis effective components can effectively prevention and cure PD,and its mechanism may be related to down regulating the expression of kisspeptin/GPR54system in hypothalamic-pituitary-ovarian axis.
作者
王瑞琼
王志旺
杜丽东
李泉洋
王晓禹
吴国泰
WANG Rui-qiong;WANG Zhi-wang;DU Li-dong;LI Quan-yang;WANG Xiao-yu;WU Guo-tai(College of Pharmacy,Gansu University of Traditional Chinese Medicine,Lanzhou 730000,GansuProvince,China;KeyLaboratory of Pharmacology and Toxicology for Traditional Chinese Medicine of Gansu Province,Gansu University of Traditional Chinese Medicine,Lanzhou 730000,GansuProvince,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2023年第5期708-712,共5页
The Chinese Journal of Clinical Pharmacology
基金
国家自然科学基金地区基金资助项目(81660653)
甘肃省青年博士基金资助项目(2021QB-074)。
关键词
当归效应组分
原发性痛经
下丘脑-垂体-卵巢轴
肿瘤转移抑制基因
G蛋白偶联受体54
Angelica sinensis effective components
primary dysmenorrhea
hypothalamic-pituitary-ovarian axis
metastasis suppressor gene
G protein-coupled receptor 54
