基于CX3CL1-CX3CR1轴探讨左归降糖解郁方对糖尿病并发抑郁症模型大鼠的神经保护作用及机制  

Neuroprotective effect and mechanism of Zuogui Jiangtang Jieyu Formula on diabetes mellitus complicated with depression model rats based on CX3CL1-CX3CR1 axis

作　　者：李萍[1] 刘洋[1] 邹蔓姝 王婷婷 郭海鹏 任廷婷 何璎 王华[3] 王宇红[1,2] LI Ping;LIU Yang;ZOU Man-shu;WANG Ting-ting;GUO Hai-peng;REN Ting-ting;HE Ying;WANG Hua;WANG Yu-hong(Technology Innovation Center,Hunan University of Chinese Medicine,Changsha 410208,China;Hunan Provincial Key Laboratory of Prevention and Treatment of Depressive Diseases with Traditional Chinese Medicine,Changsha 410208,China;the First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410007,China)

机构地区：[1]湖南中医药大学科技创新中心,湖南长沙410208 [2]抑郁类疾病中医药防治湖南省重点实验室,湖南长沙410208 [3]湖南中医药大学第一附属医院,湖南长沙410007

出　　处：《中国中药杂志》2023年第21期5822-5829,共8页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81874464,82174357);湖南省科药联合基金项目(2023JJ60476);湖南省自然科学基金青年基金项目(2022JJ40323);湖南省中医药管理局科研课题(B2023141);长沙市自然科学基金项目(kq2202266)。

摘　　要：基于CX3C趋化因子配体1(CX3C chemokine ligand 1,CX3CL1)-CX3C趋化因子受体1(CX3C chemokine receptor 1,CX3CR1)轴,探讨左归降糖解郁方改善糖尿病并发抑郁症(diabetes mellitus complicated with depression,DD)模型大鼠神经炎症及增强神经保护作用的相关机制。通过4周高脂饲料饲养联合链脲佐菌素(streptozotocin,STZ)腹腔注射及5周慢性温和不可预知应激(chronic unpredictable mild stress,CUMS)联合孤笼饲养建立DD大鼠模型,实验分为对照组、模型组、阳性药组、抑制剂组、中药组。旷场实验和强迫游泳实验评估大鼠抑郁情况,酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)检测血浆炎症因子白细胞介素-1β(interleukin-1β,IL-1β)和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的水平,免疫荧光检测海马离子钙结合衔接分子1(ionized calcium-binding adapter molecule 1,Iba1)、突触后致密蛋白-95(postsynaptic density protein-95,PSD95)、突触蛋白-1(synapsin-1,SYN1),苏木素-伊红(hematoxylin-eosin,HE)染色、尼氏(Nissl)染色和原位末端标记法(TdT-mediated dUTP nick end labeling,TUNEL)荧光染色检测海马神经元损伤情况,免疫印迹法(Western blot)检测CX3CL1-CX3CR1轴相关蛋白CX3CL1、CX3CR1、腺苷A2A受体(A2A adenosine receptor,A2AR)、谷氨酸受体2A(glutamate receptor 2A,NR2A)、谷氨酸受体2B(glutamate receptor 2B,NR2B)和脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)的表达情况。与模型组比较,左归降糖解郁方可以改善DD大鼠抑郁行为,增强神经保护作用;显著升高PSD95、SYN1、BDNF表达(P<0.01),降低Iba1、IL-1β和TNF-α表达(P<0.01)以及CX3CL1、CX3CR1、A2AR、NR2A和NR2B表达(P<0.01)。结果表明,左归降糖解郁方可能通过抑制CX3CL1-CX3CR1轴和海马小胶质细胞(microglia,MG)激活,进一步改善神经炎症与N-甲基-D-天冬氨酸受体(N-methyl-D-aspartate receptor,NMDA)亚基异常活化,从而提高海马中突触相关蛋白PSD95、SYN1和BBased on the CX3C chemokine ligand 1(CX3CL1)-CX3C chemokine receptor 1(CX3CR1) axis,this study explored the potential mechanism by which Zuogui Jiangtang Jieyu Formula(ZGJTJY) improved neuroinflammation and enhanced neuroprotective effect in a rat model of diabetes mellitus complicated with depression(DD).The DD rat model was established by feeding a high-fat diet combined with streptozotocin(STZ) intraperitoneal injection for four weeks and chronic unpredictable mild stress(CUMS) combined with isolated cage rearing for five weeks.The rats were divided into a control group,a model group,a positive control group,an inhibitor group,and a ZGJTJY group.The open field test and forced swimming test were used to assess the depression-like behaviors of the rats.Enzyme-linked immunosorbent assay(ELISA) was performed to measure the expression levels of the pro-inflammatory cytokines interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) in plasma.Immunofluorescence staining was used to detect the expression of ionized calcium-binding adapter molecule 1(Iba1),postsynaptic density protein-95(PSD95),and synapsin-1(SYN1) in the hippocampus.Hematoxylin-eosin(HE) staining,Nissl staining,and TdT-mediated dUTP nick end labeling(TUNEL) fluorescence staining were performed to assess hippocampal neuronal damage.Western blot was used to measure the expression levels of CX3CL1,CX3CR1,A2A adenosine receptor(A2AR),glutamate receptor 2A(NR2A),glutamate receptor 2B(NR2B),and brain-derived neurotrophic factor(BDNF) in the hippocampus.Compared with the model group,the ZGJTJY group showed improved depression-like behaviors in DD rats,enhanced neuroprotective effect,increased expression of PSD95,SYN1,and BDNF(P<0.01),and decreased expression of Iba1,IL-1β,and TNF-α(P<0.01),as well as the expression of CX3CL1,CX3CR1,A2AR,NR2A,and NR2B(P<0.01).These results suggest that ZGJTJY may exert its neuroprotective effect by inhibiting the CX3CL1-CX3CR1 axis and activation of hippocampal microglia,thereby improving neuroinflammation and abnormal

关 键 词：左归降糖解郁方 糖尿病并发抑郁症 CX3C趋化因子配体1(CX3CL1) CX3C趋化因子受体1(CX3CR1) 突触蛋白 

分 类 号：R285.5[医药卫生—中药学]