调更汤通过ERα/JNK/c-Jun通路对去势SD大鼠下丘脑神经细胞损伤的保护作用

Neuroprotective Effect of Tiao Geng Decoction on Hypothalamic Tissue Injury of Castrated SD Rat Through ERα/JNK/c-Jun Pathway

目的:研究调更汤通过干预雌激素受体α(estrogen receptorα,ERα),c-Jun氨基末端激酶(c-Jun N-terminal kinase,JNK),c-Jun蛋白改善去势5个月龄SD大鼠下丘脑神经细胞损伤的作用机制。方法:将40只4个月龄的SPF级SD雌性大鼠随机分为对照组、模型组、西药组、调更汤组。对照组行假手术处理,其余3组行双侧卵巢切除术,其中对照组与模型组给予生理盐水灌胃,西药组给予补佳乐(0.12 mg·kg^-1)灌胃,调更汤组给予中药调更汤[19.37 g(生药)·kg^-1]灌胃。Nissl染色法观察下丘脑神经细胞病理形态学改变;免疫荧光法检测ERα蛋白表达;免疫组织化学法检测JNK、磷酸化JNK(Phospho-c-Jun N-terminal kinase,p-JNK)、c-Jun蛋白表达。结果:对照组下丘脑神经细胞结构清晰,尼氏体分布均匀,形态正常呈“虎斑”状,与对照组比较,模型组下丘脑尼氏体减少、肥大,染色变浅;模型组下丘脑组织中ERα蛋白表达显著减少(P<0.01),p-JNK、c-Jun蛋白表达显著增加(P<0.01)。调更汤干预后,与模型组比较,调更汤组尼氏体增多,形态正常,染色加深;ERα蛋白表达显著增多(P<0.01),p-JNK、c-Jun蛋白的表达显著降低(P<0.01)。结论:调更汤可能通过干预下丘脑组织中ERα、p-JNK、c-Jun蛋白起到神经保护的作用。

Objective:To study the neuroprotective effect of Tiao Geng Decoction to relieve the injury of ovariectomized 5-month-old SD rats by interfering with estrogen receptorα(ERα),c-Jun N-terminal kinase(JNK),and c-Jun protein.The mechanism of hypothalamic nerve cell injury in aged SD rats.Methods:404-month-old SPF female SD rats were randomly divided into control group,model group,western medicine group,and TiaoGeng Decoction group.The control group underwent sham operation,and the other three groups underwent bilateral oophorectomy.The control group and the model group were given saline intragastric administration,and the western medicine group was given Progynova(0.12 mg·kg^-1)intragastric administration.The decoction group was given traditional Chinese medicine Tiao Geng Decoction[19.37 g(crude medicine)·kg^-1]by gavage.Nissl staining method to observe the pathological and morphological changes of hypothalamic nerve cells;immunofluorescence method to detect ERαprotein expression;immune-histochemical method to detect JNK,phosphorylated JNK(p-JNK),c-Jun Protein.Results:In the control group,the structure of hypothalamic nerve cells was clear,the Nissl bodies were evenly distributed,and the morphology was normal in a"tabbed"shape.Compared with the control group,the hypothalamic Nissl bodies of the model group were reduced,hypertrophy,and the staining became shallow;the expression of ERαprotein in tissues was significantly reduced(P<0.01),and the expression of p-JNK and c-Jun proteins was significantly increased(P<0.01).After Tiaogengtang intervention,compared with the model group,the Tiao Geng Decoction group had more Nissl bodies,normal morphology,and darkened staining;ERαprotein expression was significantly increased(P<0.01),and p-JNK,c-Jun protein expression was significantly reduced(P<0.01).Conclusion:Tiao Geng Decoction may play a neuroprotective effect by interfering with ERα,p-JNK and c-Jun proteins in hypothalamic tissue.