基于TLR4/MyD88/NF-κB通路介导的NLRP3炎性小体活性探讨振腹推拿改善CUMS模型大鼠海马组织炎性损伤的手法机制研究  

Based on the NLRP3 inflammasome activity mediated by TLR4/MyD88/NF-κB pathway,to investigate the manipulation mechanism of abdominal massage to improve the hippocampal inflammatory injury in CUMS model rats

作　　者：周可林 董硕 薛小娜[3] 国生[4] 魏培栋[4] 付国兵[4] 杨靖颐 ZHOU Kelin;DONG Shuo;XUE Xiaona;GUO Sheng;WEI Peidong;FU Guobing;YANG Jingyi(Beijing Hepingli Hospital,Beijing 100013,China)

机构地区：[1]北京市和平里医院康复医学科,100013 [2]北京市东城区交道口社区卫生服务中心 [3]北京中医药大学东方医院儿科 [4]北京中医药大学东方医院推拿理疗科

出　　处：《环球中医药》2024年第10期1948-1954,共7页Global Traditional Chinese Medicine

基　　金：国家自然科学基金(81574092)。

摘　　要：目的 观察振腹推拿对抑郁症模型大鼠海马组织Toll样受体4(Toll-like Receptor 4,TLR4)/髓样分化因子88(myeloiddifferentiationfactor88,MyD88)/核因子-κB(nuclear factor kappa-B,NF-κB)通路介导的NLR蛋白3(NLR protein 3,NLRP3)炎性小体活性的影响,探讨振腹推拿改善抑郁症模型大鼠海马区炎性损伤的作用机制。方法 将40只大鼠随机分为4组,采用慢性不可预见性温和应激方法复制抑郁症大鼠模型。采用离子钙接头蛋白-1(ionized calcium binding adaptor molecule-1,Iba1)免疫荧光染色检测各组大鼠海马组织海马回(cornuammonis, CA)的小胶质细胞活化程度,采用酶联免疫吸附法法检测大鼠海马组织中肿瘤坏死因子(tumor necrosis factor, TNF)-α、白细胞介素(interleukin, IL)-1β、IL-6、IL-10的含量,采用蛋白免疫印迹法和实时荧光定量PCR(Real time-PCR,RT-PCR)法检测各组大鼠海马组织中TLR4、MyD88、磷酸化核因子-κB(phospho-nuclear factor kappa-B,p-NF-κB)、NLRP3、凋亡相关斑点样蛋白(Apoptosis-associated speck-like protein containing CARD,ASC)蛋白及其基因表达。结果 (1)与正常组比较,模型组大鼠海马组织CA1、CA2、CA3、CA4区的活化小胶质细胞占比均显著升高(P<0.01)。与模型组大鼠比较,振腹组、氟西汀组大鼠海马组织CA1、CA2、CA4区的活化小胶质细胞占比均显著降低(P<0.01),氟西汀组大鼠海马组织CA3区的活化小胶质细胞占比与模型组相比降低有统计学差异(P<0.05),振腹组大鼠海马组织CA3区的活化小胶质细胞占比与模型组相比仅有降低趋势。(2)与正常组比较,模型组大鼠海马组织TNF-α、IL-1β、IL-6、含半胱氨酸的天冬氨酸蛋白水解酶-1含量显著升高(P<0.01),IL-10含量显著降低(P<0.01)。与模型组大鼠比较,振腹组、氟西汀组大鼠IL-10含量均显著升高(P<0.01),TNF-α、IL-1β、IL-6、Caspase-1含量均显著降低(P<0.01)。(3)与正常组比较,模型组大鼠海马组织TLR4、MyD88、p-NF-κBObjective To observe the effect of abdominal massage on the activity of NLR protein 3(NLRP3)inflammasome mediated by toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor-κB(NF-κB)pathway in hippocampus of rats with depression.To explore the mechanism of abdominal massage to improve the hippocampal inflammatory damage in rats with depression.Methods 40 rats were randomly divided into 4 groups and the depression rat model was reproduced by chronic unpredictability mild stress.Iba1 immunofluorescence staining was used to detect the activation degree of microglia in hippocampal CA region of rats,and ELISA was used to detect the contents of tumor necrosis factorα(TNF-α),interleukin-1β(IL-1β),white hormone 6(IL-6)and interleukin-10(IL-10)in hippocampal tissue of rats.Western blot and real-time quantitative PCR were used to detect the expression of TLR4,MyD88,p-NF-κB,NLRP3 and ASC proteins and their genes in hippocampal tissues of rats in each group.Results(1)Compared with the normal group,the proportion of activated microglia in CA1,CA2,CA3 and CA4 regions of hippocampus in the model group was significantly increased(P<0.01).Compared with the model group,the proportion of activated microglia in CA1,CA2 and CA4 regions of hippocampal tissue of rats in the abdominal oscillation group and the fluoxetine group were significantly decreased(P<0.01),the proportion of activated microglia in CA3 region of hippocampal tissue in the fluoxetine group was significantly lower than that in the model group(P<0.05),compared with the model group,the proportion of activated microglia in CA3 region of hippocampal tissue of rats in the shaking group had a decreasing trend.(2)Compared with the normal group,the contents of TNF-α,IL-1β,IL-6 and Caspase-1 in hippocampus of rats in the model group were significantly increased(P<0.01),IL-10 content was significantly decreased(P<0.01).Compared with the model group,the IL-10 content of rats in the abdominal oscillation group and the fluoxetine group was signi

关 键 词：振腹推拿 抑郁症 海马 炎性损伤 Toll样受体4/髓样分化因子88/核因子-κB通路 NLR蛋白3炎性小体 小胶质细胞活化 

分 类 号：R285.5[医药卫生—中药学]