柴胡疏肝散调节SIK2-CRTC1通路改善围绝经期肝郁证的机制  

Mechanism of Chaihu Shugan San on regulating the SIK2-CRTC1 pathway to improve liver depression syndrome in perimenopausal mice

作　　者：王素英 蔡晓诗 陈朝玺 钟瑶 李欣媛 沈建英[1,2,3] 龚琳 梁文娜 WANG Suying;CAI Xiaoshi;CHEN Chaoxi;ZHONG Yao;LI Xinyuan;SHEN Jianying;GONG Lin;LIANG Wenna(School of Chinese Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;Fujian Province Collaborative Innovation Center of TCM Health Management in 2011,Fuzhou 350122,China;Key Laboratory of TCM Health Status Identification of Fujian Province,Fuzhou 350122,China)

机构地区：[1]福建中医药大学中医学院,福州350122 [2]福建省2011中医健康管理协同创新中心,福州350122 [3]福建省中医健康状态辨识重点实验室,福州350122

出　　处：《中华中医药杂志》2024年第11期5869-5874,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.82274409,No.82174248);福建省自然科学基金项目(No.2023J01868)。

摘　　要：目的:基于盐诱导激酶2(SIK2)-CREB调节转录共激活因子1(CRTC1)通路探讨柴胡疏肝散改善围绝经期肝郁证的相关机制。方法:采用去势+慢性温和不可预知性应激(CUMS)法建立围绝经期肝郁证模型。24只雌性C57BL/6小鼠按体质量分层随机分为假手术组、围绝组、围绝肝郁组和柴胡疏肝散组,每组6只。通过旷场实验(OFT)、强迫游泳实验(FST)和糖水偏好实验检测小鼠行为学变化;ELISA法测定小鼠血清雌二醇(E2)、促卵泡素(FSH)、5羟色胺(5-HT)和多巴胺(DA)水平;尼氏染色观察海马神经元形态结构;Western blot法检测海马SIK2、CRTC1、磷酸化CRTC1(pCRTC1)、环磷酸腺苷反应元件结合蛋白(CREB)和脑源性神经营养因子(BDNF)的蛋白相对表达量;免疫荧光标记法检测海马突触后致密蛋白95(PSD95)和缝隙连接蛋白36(Cx36)表达。结果:与围绝肝郁组比较,柴胡疏肝散组小鼠的焦虑、抑郁样行为明显改善,FST不动时间显著缩短(P<0.05),1%蔗糖水偏好率显著升高(P<0.01);血清FSH水平显著降低,E2、5-HT和DA水平显著升高(P<0.01);海马神经元损伤程度减轻;SIK2-CRTC1通路相关蛋白表达被不同程度逆转,以pCRTC1表达降低最为显著(P<0.01);PSD95、Cx36蛋白阳性表达亦显著提高(P<0.01,P<0.05)。结论:柴胡疏肝散可在一定程度上调控SIK2-CRTC1通路的表达,减轻海马神经元损伤,改善围绝经期肝郁证小鼠焦虑抑郁样行为。Objective:To explore the mechanisms underlying the improvement of liver depression syndrome in perimenopausal mice by Chaihu Shugan San(CSS)based on the salt-inducible kinase(SIK2)-CREB regulated transcription coactivator 1(CRTC1)pathway.Methods:The perimenopausal liver depression syndrome model was established using the ovariectomy+chronic unpredictable mild stress(CUMS)method.A total of 24 female C57BL/6 mice were randomly divided into sham surgery group,perimenopausal group,perimenopausal liver depression group,and CSS group,with 6 mice in each group.Behavioral changes of mice were assessed by open field test(OFT),forced swimming test(FST),and 1%sucrose preference test(SPT).Serum levels of estradiol(E2),follicle-stimulating hormone(FSH),5-hydroxytryptamine(5-HT),and dopamine(DA)were measured by ELISA.Nissl staining was performed to observe the morphological structure of hippocampal neurons.Western Blot was used to detect the expression of SIK2,CRTC1,pCRTC1,AMP responsive element binding protein(CREB),and brain derived neurotrophic factor(BDNF)in the hippocampus.Immunofluorescence labeling was conducted to detect the expression of postsynaptic density protein 95(PSD95)and connexin 36(Cx36)in the hippocampus.Results:Compared with the perimenopausal liver depression group,mice in the CSS group showed significant improvement in anxiety and depression-like behaviors,with a significant shorteing of the FST immobility duration(P<0.05)and a marked increase in the preference for 1%sucrose water(P<0.01);serum FSH levels were significantly decreased,while E2,5-HT,and DA levels were significantly increased(P<0.01);the degree of hippocampal neuron damage was reduced;the expression of SIK2-CRTC1 pathway-related proteins was reversed to varying degrees,with the most significant decrease observed in pCRTC1 expression(P<0.01);positive expression of PSD95 and Cx36 proteins were also significantly increased(P<0.01,P<0.05).Conclusion:CSS could alleviate anxiety and depression-like behaviors in perimenopausal liver depression synd

关 键 词：围绝经期综合征 肝郁证 盐诱导激酶2(SIK2)-CREB调节转录共激活因子1(CRTC1) 神经元 柴胡疏肝散 

分 类 号：R285.5[医药卫生—中药学]