西黄丸对大鼠卵巢相关激素的基因和蛋白表达水平的影响

Effect of Xihuang pill on the expression levels of ovarian hormone gene and protein in rats

目的研究西黄丸对大鼠卵巢雌二醇、孕激素、卵泡刺激素受体(FSHR)的基因和蛋白、促黄体生成素受体(LHR)基因和蛋白的影响及作用机制。方法按随机数字表法将雌性未孕Wistar大鼠随机分为5组:空白组(灌胃等体积的蒸馏水)、对照组(灌胃1. 8 mg·kg-1·d-1他莫昔芬)和高、中、低3个剂量实验组(灌胃2. 16,1. 08,0. 54 g·kg-1·d-1西黄丸),每组10只。以酶联免疫法检测卵巢雌二醇和孕激素含量;用实时荧光定量聚合酶链反应(PCR)法和蛋白免疫印迹技术分别检测FSHR基因、LHR基因和FSHR蛋白、LHR蛋白的表达量。结果空白组、对照组和高、中、低3个剂量实验组的雌二醇含量分别为(45. 85±17. 39),(19. 45±6. 41),(28. 74±14. 54),(29. 82±11. 57)和(44. 11±8. 25) ng·L-1;这5组的孕激素含量分别为(259. 67±12. 31),(301. 21±15. 01),(296. 47±14. 04),(301. 56±22. 81)和(287. 13±14. 16)pmol·L-1;这5组的FSHR蛋白相对表达量(光密度值)分别为1. 31±0. 11,0. 90±0. 04,0. 41±0. 01,0. 86±0. 02和1. 31±0. 02;这5组的LHR蛋白相对表达量(光密度值)分别为0. 87±0. 05,0. 91±0. 04,3. 01±0. 13,1. 96±0. 00和1. 21±0. 02,对照组和高、中2个剂量实验组与空白组比较,上述指标的差异均有统计学意义(均P <0. 01)。基因结果的趋势与蛋白一致。结论西黄丸能使大鼠卵巢中雌二醇的含量降低,能使孕激素的含量增加,对FSHR基因和FSHR蛋白的表达有抑制作用,对LHR基因和LHR蛋白的表达有促进作用。说明西黄丸能干预大鼠的卵巢功能。

Objective To investigate the effect of Xihuang pill on estradiol,progesterone,follicle stimulating hormone receptor(FSHR)gene and protein,luteinizing hormone receptor(LHR)gene and protein in rat ovary,and its mechanism.Methods Non-pregnant Wistar female rats were randomly divided into five groups:blank group(given distilled water of equal volume),control group(given 1.8 mg·kg-1·d-1 tamoxifen),and high,medium and low dose experimental groups(given Xihuang pill of 2.16,1.08,0.54 g·kg-1·d-1),with 10 rats in each group.The contents of estradiol and progesterone in ovaries were detected by enzyme linked immunosorbent assay.The expressions of FSHR,LHR gene and FSHR,LHR protein were detected by real-time fluorescence quantitative PCR and Western blot,respectively.Results The estradiol in blank group,control group,and high,medium and low dose experimental groups were respectively(45.85±17.39),(19.45±6.41),(28.74±14.54),(29.82±11.57),and(44.11±8.25)ng·L-1;the progesterone in the 5 groups were respectively(259.67±12.31),(301.21±15.01),(296.47±14.04),(301.56±22.81),and(287.13±14.16)pmol·L-1;the relative expressions of FSHR protein of these 5 groups were respectively1.31±0.11,0.90±0.04,0.41±0.01,0.86±0.02,and 1.31±0.02;the relative expressions of LHR protein in the 5 groups were respectively 0.87±0.05,0.91±0.04,3.01±0.13,1.96±0.00,and 1.21±0.02.The differences of the factors between the control group,high and medium dosage experimental groups with blank group were statistically significant(all P<0.01).The trend of gene results is consistent with that of protein.Conclusion Xihuang pill can reduce estradiol content in rat ovary,promote progesterone content,inhibit expression of FSHR gene and FSHR protein,and promote expression of LHR gene and LHR protein,which indicates that Xihuang pill can interfere with ovarian function.