莉芙敏对去卵巢大鼠下丘脑视前区5-HT1AR和5-HT2AR表达的影响

Effect of Remifemin on Expression of 5-HT1AR and 5-HT2AR in Preoptic Area of Hypothalamus of Ovariectomized Rats

目的:观察去卵巢大鼠应用黑升麻异丙醇提取物——莉芙敏(ICR)治疗1～4周后,免疫组化方法检测5-羟色胺1A受体(5-HT1AR)和5-HT2AR在大鼠下丘脑视前区表达的变化情况,为莉芙敏缓解围绝经期潮热症状的机制研究提供形态学依据。方法:雌性大鼠分为假手术组(Sham组)、去卵巢组(OVX组)、OVX后戊酸雌二醇治疗(OVX+E2)组和OVX后ICR治疗(OVX+ICR)组,每组40只。所有大鼠术后恢复2周后,再用相应的药物分别治疗1,2,3,4周,麻醉,心脏灌注,取脑,制作冰冻切片。免疫组化法观察各组大鼠下丘脑视前区5-HT1AR和5-HT2AR的表达。结果:①OVX组1～4周时大鼠下丘脑视前区5-HT1AR阳性细胞数量和吸光度均较同期Sham组增加;ICR和戊酸E2治疗1～4周后,OVX+E2组和OVX+ICR组大鼠下丘脑视前区室周带表达5-HT1AR的阳性细胞数量和吸光度均较同期OVX组减少。②下丘脑视前区中间带和外侧区5-HT2AR阳性细胞数量和吸光度的变化:OVX组大鼠,1～4周均较同期Sham组增加;ICR和戊酸E2治疗1周和2周后,OVX+E组和OVX+ICR组较同期OVX组增加;治疗3周和4周后,OVX+E2组和OVX+ICR组均较同期OVX组减少。结论:5-HT1AR和5-HT2AR在去卵巢大鼠下丘脑视前区的表达量均增加;ICR和戊酸E2治疗后,5-HT1AR在大鼠下丘脑视前区的表达量减少,而5-HT2AR在下丘脑视前区的表达先增多后减少。ICR可能通过调节下丘脑视前区体温调节中枢的5-HT1AR和5-HT2AR的表达,以缓解围绝经期的潮热症状。

Objective：To investigate regional changes in 5-hydroxytryptamine（5-HT）1A receptors（5-HT1AR） and 5-hydroxytryptamine（5-HT） 2A receptors（5-HT2AR） in the OVX rat preoptic area of hypothalamus after 1-4 weeks treatment with isopropanolic black cohosh extract—remifemin（ICR）,and explore its potential mechanism of alleviating hot flushes during the peri-menopausal period.Methods：The rats were divided into four groups,ovariectomy（OVX） group,sham group,ovariectomy with estradiol valerate（E2） treatment（OVX＋E2）group,and ovariectomy with ICR treatment（OVX＋ICR） group.The sham group underwent a sham operation without ovariectomies,while the other three groups underwent bilateral ovariectomies under sterile conditions.All rats were given 2 weeks to recover after the operation,then each group was given a different treatment lasting 1 week,2 weeks,3 weeks and 4 weeks.The expression of 5-HT1AR and 5-HT2AR in the rat preoptic area of hypothalamus（POAH）were detected by immunohistochemistry.Results：①After 1-4 weeks＇ treatment,the number of 5-HT1AR positive cells and the optical density（OD） in POAH of OVX group significantly increased compared with that in the same period of sham group;while the number of 5-HT1AR positive cells and the optical density in periventricular zone of OVX＋ICR group and OVX＋E2 group significantly decreased compared with that in the same period of OVX group.②After 1-4 weeks＇ treatment,the number of 5-HT2AR positive cells and the optical density（OD） in intermediate zone and lateral zone of OVX group significantly increased compared with that in the same period of sham group.The number of 5-HT2AR positive cells and the optical density（OD） in intermediate zone and lateral zone of OVX＋ICR group and OVX＋E2 group significantly increased compared with that in the same period of OVX group after 1-2 weeks＇ treatment.While the result is reverse after 3-4 weeks＇ treatment.Conclusions：Ovariectomized rats had increased expression of 5-HT1AR and 5-HT2AR.After treatment with remifemin or estradiol valerate,the expression of 5-HT1AR in POAH significantly decreased.While the expression of 5-HT2AR in POAH decreased following an increased period.Regulating the expression of 5-HT1AR and 5-HT2AR in POAH may be one of the mechanisms of remifemin alleviating menopausal hot flush.