西黄丸对大鼠垂体相关激素的基因与蛋白表达水平的影响

Effects of Xihuang Pill on genes and proteins expression levels of pituitary-related hormones in rats

目的探讨西黄丸对大鼠垂体卵泡刺激素(FSH)、促黄体生成素(LH)、促性腺激素释放激素受体(GnRHR)基因和GnRHR蛋白的影响。方法按照随机数表法将Wistar雌性未孕大鼠随机分为5组:空白组、对照组和高、中、低3个剂量实验组,每组10只。对照组每只大鼠按1.8 mg·kg^-1·d-1他莫昔芬灌胃;高、中、低3个剂量实验组分别给予2.16,1.08,0.54 g·kg^-1·d-1西黄丸灌胃;空白组灌胃等体积蒸馏水。以酶联免疫吸附法检测垂体FSH和LH含量;以蛋白免疫印迹技术检测垂体中GnRHR蛋白表达量;以实时荧光定量PCR检测GnRHR基因表达量。结果空白组、对照组和高、中、低3个剂量实验组的FSH含量分别为(46.88±15.66),(12.23±4.58),(20.23±7.37),(28.23±5.39)和(35.70±8.51)U·L^-1;上述这5组的LH含量分别为(48.72±3.03),(59.77±2.38),(55.71±4.56),(61.51±3.19)和(52.80±3.79)ng·L^-1;上述这5组的GnRHR蛋白表达水平(OD值)分别为4.45±0.05,1.86±0.04,1.95±0.05,2.23±0.02和2.31±0.05。上述指标:对照组和3个剂量实验组与空白组比较,差异均有统计学意义(P<0.05或P<0.01)。基因结果的趋势与蛋白一致。结论西黄丸能够对垂体的功能产生干预作用。

Objective To investigate the effects of Xihuang Pill on the pituitary follicle stimulating hormone(FSH),luteinizing hormone(LH),gonadotropin-releasing hormone receptor(GnRHR)gene,and GnRHR protein.Methods Non-pregnant Wistar female rats were randomly divided into five groups according to random number table:blank group,control group,as well as low,medium and high dose of experimental groups,with 10 rats in each group.Each rat in the control group was given tamoxifen according to 1.8 mg·kg^-1·d-1,each rat in the high,medium and low dose groups was given Xihuang Pill according to 2.16,1.08,0.54 g·kg^-1·d-1,and the blank group was given distilled water of equal volume.The contents of FSH and LH in pituitary gland were detected by enzyme linked immunosorbent assay.The expression of GnRHR protein in pituitary was detected by Western blot.The expression of GnRHR gene were detected by real-time fluorescence quantitative-PCR.Results The FSH content in blank group,control group,experimental-H,experimental-M,experimental-L groups were respectively(46.88±15.66),(12.23±4.58),(20.23±7.37),(28.23±5.39),(35.70±8.51)U·L^-1;the LH content in the above 5 groups were respectively(48.72±3.03),(59.77±2.38),(55.71±4.56),(61.51±3.19),(52.80±3.79)ng·L^-1;the expression of GnRHR protein(OD value)in the above 5 groups were respectively4.45±0.05,1.86±0.04,1.95±0.05,2.23±0.02,2.31±0.05.Comparison between the control group,experimental-H,experimental-M,experimental-L groups and blank group,the difference of the factors were statistical significance(P<0.05 or P<0.01).The trend of gene results is consistent with that of protein.Conclusion Xihuang Pill can interfere with the fuction of pituitary.