温阳、解郁及温阳解郁方对母婴分离结合慢性神经疼痛应激小鼠疼痛敏感性及抑郁样行为的调节作用  

Regulatory Effect of Wenyang Prescription,Jieyu Prescription,and Wenyang Jieyu Prescription on Pain Sensitivity and Depression-like Behaviors in Mice Induced by Maternal Separation and Chronic Neuropathic Pain

作　　者：左阳 赵永烈 巩子汉 孟丹华 佘楷杰 张艺嘉 梁文青 楚天云 岳广欣[3] ZUO Yang;ZHAO Yonglie;GONG Zihan;MENG Danhua;SHE Kaijie;ZHANG Yijia;LIANG Wenqing;CHU Tianyun;YUE Guangxin(Beijing University of Chinese Medicine,Beijing 100029,China;Third Affiliated Hospital,Beijing University of Chinese Medicine,Beijing 100029,China;Institute of Basic Theories of Traditional Chinese Medicine(TCM),China Academy of Chinese Medical Sciences,Beijing 100700,China;TCM School,Henan University of Chinese Medicine,Zhengzhou 453000,China;Henan Provincial Orthopedic Hospital,Zhengzhou 453000,China)

机构地区：[1]北京中医药大学,北京100029 [2]北京中医药大学第三附属医院,北京100029 [3]中国中医科学院中医基础理论研究所,北京100700 [4]河南中医药大学中医学院,郑州453000 [5]河南省骨科医院,郑州453000

出　　处：《中国实验方剂学杂志》2022年第14期44-53,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上项目(82174251,81573846);中国中医科学院科技创新工程重大攻关项目(CI2021A00607);中央级公益性科研院所基本科研业务费专项资金资助自主选题项目(YZ1883,YZ202047,YZ-202107,YZ-202148)。

摘　　要：目的:观察母婴分离小鼠给予慢性神经疼痛应激后的行为、痛阈改变,以及血清和中枢抑郁、疼痛相关指标的变化,以温阳方、解郁方、及温阳解郁方进行干预探讨温阳解郁法抗抑郁、改善疼痛敏感的可能机制。方法:以小鼠出生日为PD0,小鼠在出生后随机分为对空白组和母婴分离组,母婴分离组在PD5~14进行8 h·d;的母婴分离,离乳后将其分为母婴分离组、温阳组、解郁组、温阳解郁组和氟西汀组,每组15只,同时以15只未经母婴分离的同期雄性仔鼠作为空白组。空白组、母婴分离组小鼠于PD21~90以普通发育饲料喂养,余组分别以温阳方二仙汤、解郁方逍遥散、温阳解郁方及氟西汀药混饲料喂养(用药剂量分别为5.84、12.00、16.68 g·kg;及2.60 mg·kg;)。于PD91行坐骨神经结扎手术,建立母婴分离+慢性神经病理性疼痛模型。采用旷场实验观察各组小鼠抑郁样行为,测量机械痛阈和温度痛阈检测各组小鼠疼痛敏感性,酶联免疫吸附测定法(ELISA)检测血清皮质酮(CORT)、P物质及β-内啡肽(β-EP)含量,免疫组化检测杏仁核糖皮质激素受体(GR)和下丘脑β-EP蛋白表达,实时荧光定量聚合酶链式反应(Real-time PCR)检测杏仁核糖皮质激素受体基因(Nr3c1)、FK506结合蛋白5基因(FKBP5)、代谢型谷氨酸受体5基因(GRM5)及脑源性神经营养因子(BDNF)mRNA的表达量。结果:与空白组比较,母婴分离组小鼠5 min旷场中心停留时间和运动总距离均显著减少(P<0.01),机械痛阈显著降低(P<0.01)。血清CORT和β-EP均显著上升(P<0.01),FKBP5 mRNA表达显著下降(P<0.01)。下丘脑β-EP表达上升,差异具有明显统计学意义(P<0.05)。与母婴分离组比较,各给药组5 min旷场中心停留时间均明显增加(P<0.05,P<0.01),温阳组、温阳解郁组和氟西汀组小鼠在5 min旷场运动总距离明显增加(P<0.05,P<0.01),各给药组痛阈均不同程度上调。解郁组小鼠血清CORT显著下降(PObjective:To observe the behavioral and pain threshold alterations,as well as the changes in indexes related to depression and pain in the serum and central system in mice stressed by maternal separation and chronic neuropathic pain,and explore the underlying mechanism of Wenyang prescription(WY),Jieyu prescription(JY),and Wenyang Jieyu prescription(WYJY)in improving depression and pain sensitivity.Method:The birth date of mice was recorded as PD0. After birth,the mice were divided into a blank group and an experimental group. The neonatal mice in the experimental group underwent maternal separation in PD5-14 at8 h·d;. After ablactation,the mice were divided into a maternal separation group,a WY group(Erxian decoction,5.84 g·kg;),a JY group(Xiaoyaosan,12.00 g·kg;),a WYJY group(16.68 g·kg;),and a fluoxetine group(2.60 mg·kg;),with 15 mice in each group. Meanwhile,15 male mice of the same age without maternal separation were assigned to the normal control group. Mice in the blank group and the maternal separation group were fed on a regular chow diet in PD21-PD90,while the remaining groups were fed on the corresponding drugs. In PD91,sciatic nerve ligation was performed to induce a model of maternal separation and chronic neuropathic pain. The open field test was used to observe the depression-like behaviors of mice in each group,and the mechanical and temperature pain thresholds were measured to detect the pain sensitivity of mice in each group. The serum levels of corticosterone(CORT),substance P,and β-endorphin(β-EP)were determined by enzyme-linked immunosorbent assay(ELISA),and the expression of the glucocorticoid receptor(GR)in the amygdala and β-EP protein in the hypothalamus was detected by immunohistochemistry. The mRNA expression levels of amygdala GR gene(Nr3c1),FK506 binding protein 5 gene(FKBP5),metabolic glutamate receptor 5 gene(GRM5), and brain-derived neurotrophic factor(BDNF) were detected by real-time fluorescence quantitative polymerase chain reaction(Real-time PCR). Result: Compared with

关 键 词：母婴分离 温阳解郁方 杏仁核 慢性神经疼痛 抑郁 

分 类 号：R2-0[医药卫生—中医学] R33[哲学宗教—心理学] R289[医药卫生—医学心理学] R395.3