四逆散对抑郁大鼠NLRP3炎症小体及抑郁样行为的作用  

Effect of Sinisan on NLRP3 Inflammasomes and Depression-like Behaviors in Depressed Rats

作　　者：王威[1] 周艳艳[1] 喻小明[1] 江晓翠 田代志[1] 萧闵[1] WANG Wei;ZHOU Yan-yan;YU Xiao-ming;JIANG Xiao-cui;TIAN Dai-zhi;XIAO Min(Hubei University of Chinese Medicine,Wuhan 430065,China)

机构地区：[1]湖北中医药大学,武汉430065

出　　处：《中国实验方剂学杂志》2022年第12期22-30,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(82104704,81703954)。

摘　　要：目的观察四逆散对慢性温和不可预知应激(CUMS)诱导的抑郁症大鼠行为学及NOD样受体热蛋白结构域3(NLRP3)炎症小体的作用,进一步探讨四逆散抗抑郁的作用机制。方法将50只雄性大鼠随机分为正常组、模型组、NLRP3抑制剂(MCC950)组、四逆散低、高剂量,每组10只。除正常组外,均采用为期42 d的CUMS诱导各组大鼠抑郁。造模第22天开始药物干预,四逆散低、高剂量每天以四逆散(2.5、5 g·kg^(-1))进行灌胃,MCC950组以每天腹腔注射MCC950(10 mg·kg^(-1))。同时,除MCC950组外,余下4组腹腔注射10 mg·kg^(-1)生理盐水;而模型组、正常组、MCC950组灌胃3 mL生理盐水。21 d后进行糖水偏好实验、旷场实验;蛋白免疫印迹法(Western blot)检测各组大鼠海马组织NLRP3、相关斑点样蛋白(ASC)、胱天蛋白酶-1(Caspase-1)、B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、离子钙结合衔接蛋白1(Iba1)、CD68蛋白表达量。酶联免疫吸附测定法(ELISA)检测大鼠海马组织白细胞介素-1β(IL-1β)、白细胞介素-18(IL-18)含量。尼氏染色评估大鼠海马CA1区病理改变,原位末端标记法(TUNEL)检测大鼠CA1区凋亡水平。结果与正常组比较,模型组大鼠糖水偏好率及消耗体积,旷场实验中总距离、中央区运动距离百分比和停留时间百分显著下降(P<0.01);与模型组比较,四逆散高、低剂量组和MCC950组大鼠抑郁行为,海马CA1区凋亡水平及神经元丢失不同程度的减轻。四逆散可明显降低海马组织IL-1β、IL-18、Bax、Iba1、CD68水平(P<0.05,P<0.01),上调Bcl-2水平(P<0.05,P<0.01),并且抑制NLRP3炎症小体相关蛋白NLRP3、ASC、Caspase-1的表达(P<0.05,P<0.01)。结论四逆散可改善CUMS大鼠的抑郁样行为、海马区神经元病理性损伤,其机制可能与NLRP3炎症小体信号通路介导的炎症反应有关。Objective To observe the effects of Sinisan on behaviors and NOD-like receptor protein 3(NLRP3)inflammasomes of depressed rats induced by chronic unpredictable mild stress(CUMS)and further explore the anti-depressant mechanism of Sinisan.Method Fifty male rats were randomly divided into a normal group,a model group,an NLRP3 inhibitor(MCC950)group(10 mg·kg^(-1)),and low-(2.5 g·kg^(-1))and high-dose(5 g·kg^(-1))Sinisan groups,with 10 rats in each group.The depression model was induced by 42 d CUMS in rats except for those in the normal group.Drug intervention was performed on the 22^(nd) day of modeling by gavage in the Sinisan groups and by intraperitoneal injection in the MCC950 group.Except for the MCC950 group,the remaining four groups received 10 mg·kg^(-1)physiological saline by intraperitoneal injection,while the rats in the model group,the normal group,and the MCC950 group were administered with 3 mL of physiological saline by gavage.Twenty-one days later,the sucrose preference test and open field test were performed.Western blot was used to detect the protein expression of NLRP3,apoptosis-associated speck-like protein containing a CARD(ASC),cysteinyl aspartate-specific protease-1(Caspase-1),B-cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax),ionized calcium-binding adapter molecule 1(Iba1),and CD68 in the hippocampus of rats in each group.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of interleukin-1β(IL-1β)and interleukin-18(IL-18)in the hippocampus of rats.Nissl staining and TUNEL were used to assess the pathological changes and apoptosis level in the hippocampal CA1 region of rats,respectively.Result The sucrose preference rate and consumption volume in the sucrose preference test,the total distance,the percentage of central movement distance,and the percentage of residence time in the open field test of rats in the model group were lower than those in the normal group(P<0.01).Compared with the model group,the Sinisan groups and the MCC950 group showed improved depress

关 键 词：四逆散 慢性温和不可预知应激(CUMS) 神经炎症 核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎症小体 抑郁症 凋亡 

分 类 号：R2-0[医药卫生—中医学] R22