经前舒颗粒对PMS肝气郁证大鼠下丘脑ERα,ERβ分布与表达的影响

Effects of Jingqianshu Granule on Distribution and Expression of Estrogen Receptor α and β in the Hypothalamus of Premenstrual Syndrome Model Rats with Liver-qi Stagnation

目的:观察经前舒颗粒对经前期综合征(PMS)肝气郁证模型大鼠下丘脑不同核团雌激素受体(ER)α,ERβ蛋白分布及表达的影响,研究经前舒颗粒治疗该病症的中枢作用机制。方法:将30只雌性SD大鼠随机分为正常对照组、模型组、模型给药组,每组10只。采用慢性束缚应激造模法复制PMS肝气郁证大鼠模型。造模同时,模型给药组按10 g.kg-1剂量ig给药。造模结束后,通过免疫组织化学技术ABC法检测各组大鼠雌激素受体α,β在下丘脑中的定位分布并分析其积分吸光度(IA);HPLC检测各组大鼠下丘脑中5-羟色按(5-HT)含量。结果:3组大鼠雌激素受体ERα,ERβ主要分布在下丘脑的腹内侧区(VMH)核团,分布区域无差异。模型组VMH区ERα,ERβIA分别为163 605±27 284和147 318±24 924与正常组IA(130 156±32 579,105 430±28 641)相比显著升高(P<0.05,P<0.01);给药组ERα,ERβIA为122 020±21 287和77 482±31112,与模型组相比显著降低(P<0.01,P<0.01)。模型组大鼠下丘脑5-HT含量(454.51±65.18)μg.L-1与正常组(144.47±42.20)μg.L-1相比显著升高(P<0.01),给药组(234.11±14.38)μg.L-1与模型组相比5-HT含量显著下降(P<0.01)。结论:下丘脑VMH区ERα,ERβ蛋白表达升高可能是PMS肝气郁证发病的中枢机制之一。经前舒颗粒可有效改善这种状态,提示下丘脑ERα,ERβ可能是该药治疗PMS肝气郁证的部分作用靶点。

Objective： To observe the effects of Jingqianshu granule on distribution and expression of estrogen receptor （ER） a and ERE in the hypothalamus of premenstrual syndrome model rats with liver-qi stagnation, and to find out the corresponding effect targets and mechanisms of Jingqianshu granule. Method ： Thirty female SD rats were randomly divided into three groups： the normal group, the model group, and the medication group. The premenstrual syndrome （PMS） model rats with Liver-qi stagnation were induced by bondage stimulation. Jingqianshu granule was given to rats in the medication group at the dose of 10 g .kg-~ by gastrogavagewhile equal volume of sterilized water was given to modeling, rats in the model group and the normal group, once daily. The protein distribution and integrated optical density （A） in ventromedial hypothalamic area （VMH） of rats in the three groups were detected by using immunohistochemistry ABC methods. HPLC method was used to detect 5-HT level in hypothalamus. Result： IHC demonstrated that there were no noticeable differences in the distribution of ERα, ERβ in VMH of hypothalamus in three groups. The IA of ERα, ERβ in model group which were 163 605 ± 27 284 and 147 318 ± 24 924 increased markedly compared with the normal group （130 156± 32 579,105 430±28 641） （P〈0.05, P〈0.01）. The IA of ERα and ERβof medication group which were 122 0120 ±21 287 and 77 482 ± 31 112 decreased markedly compared with model group （all P 〈 0. O1 ）. The 5-HT level （454.51±65.18）μg·L^-1 in hypothalamus of model group were noticeable increased compared with the normal group （144.47±42.20） μg·L^-1 （P〈0.01）, and the 5-HT level （234.11 ＋14.38）μg·L^-1of medication group decreased markedly compared with model group （P 〈 0.01 ）. Conclusion： The high expressions of the ERα and ERβ protein in VMH may have close relationship with liver-qi stagnation, and probably is one of the centre pathogenesis of PMS with liver-qi stagnation. The Jingqianshu granule can correct the high expression of ERα and ERβ protein of model rats in VMH, which may be one of the mechanism and effect target for Jingqianshu granule.