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目的研究左归饮对围绝经期大鼠卵巢P450芳香化酶(P450 arom)及CYP19 mRNA表达的影响及机制。方法采用围绝经期大鼠动物模型,分别给予低、中、高剂量左归饮灌胃处理8周。用放射免疫法(RIA)检测血清中雌二醇(E_2)含量;用RT-PCR和免疫组化法检测卵巢CYP19mRNA及P450 arom的表达。未成年大鼠注射孕马血清后收集培养颗粒细胞,用RT-PCR方法检测给予不同处理后CYP19 mRNA的表达水平。结果围绝经期大鼠血清E_2水平及卵巢P450 arom及其基因CYP19的表达明显低于青年组(P<0.01),给予中、高剂量左归饮处理后,血清E_2水平及P450 arom、CYP19表达显著升高(P<0.01)。培养颗粒细胞给予不同处理因素,结果显示,与对照组相比,染料木黄酮(Genistein)、福司考林(Forskolin)、左归饮提取物可促进CYP19 mRNA表达(P<0.05);与单纯加入左归饮提取物相比,同时加入左归饮提取物和佛波酯或Genistein可进一步促进CYP19 mRNA的表达(P<0.05)。结论中、高剂量左归饮可通过上调卵巢CYP19及其产物P450 arom的表达,使围绝经期大鼠血清E_2含量升高。左归饮对CYP19 mRNA表达的促进作用可能是通过激活PKC途径或加强Genistein的作用实现的。
Objective To study the effect and mechanism of Zuoguiyin on the expression of P450 arom and CYP19 mRNA in the ovary of perimenopausal rats. Methods Peri-menopausal rat model was established. The rats were treated with low, medium and high doses of Zuogui Yin for 8 weeks. The content of estradiol (E2) in serum was detected by radioimmunoassay (RIA). The expression of CYP19mRNA and P450 arom in ovary was detected by RT-PCR and immunohistochemistry. The immature rats were injected with pregnant and mare serum collected after cultured granulosa cells, using RT-PCR method to detect the different treatment of CYP19 mRNA expression levels. Results The levels of serum E_2 and P450 arom gene and CYP19 in ovary were significantly lower than those in young group (P <0.01). After middle and high doses of Zuogui Yin were treated, the levels of E_2 and the expressions of P450 arom and CYP19 Significantly increased (P <0.01). The results showed that Genistein, Forskolin and Zuoguiyin extracts could promote the expression of CYP19 mRNA (P <0.05). Compared with the control group, Compared with Zuoguiyin extract, the addition of Zuoguiyin extract and phorbol ester or Genistein can further promote the expression of CYP19 mRNA (P <0.05). Conclusion The medium and high doses of Zuoguiyin can increase the content of E_2 in perimenopausal rats by up-regulating the expression of CYP19 in ovary and P450 arom. The effect of Zuoguiyin on CYP19 mRNA expression may be through activation of PKC pathway or by strengthening the effect of genistein.