摘要
目的研究低聚原花青素(oligomeric proanthocyanidins,OPCs)调控Toll样受体4 (TLR4)/p38丝裂原活化蛋白激酶(p38 MAPK)/核因子-κB (NF-κB)信号通路相关蛋白质表达保护过度训练大鼠脾脏的作用机制。方法雄性Wistar大鼠分为对照组(C组,n=12)、过度训练组(OM组,n=12)、OPCs+过度训练组(OOM组,n=13)。C组不进行任何运动干预,OM组、OOM组大鼠进行6w递增负荷游泳训练。训练期间,OOM组以5 ml/kg (150 mg/kg bw·d)OPCs灌胃,其他组灌胃等体积蒸馏水。末次训练后24 h取材,称重计算脾脏指数,HE染色观察脾脏组织病理学变化,Tunel法检测脾脏组织细胞凋亡水平,免疫组化法检测脾脏p38 MAPK、磷酸化p38 MAPK (p-p38 MAPK)、NF-κB、B淋巴细胞瘤因子-2 (Bcl-2)和Bcl-2相关X蛋白(Bax)蛋白质表达水平,流式细胞术检测单核细胞表面TLR4表达率,酶联免疫吸附法检测脾脏肿瘤坏死因子-α(TNF-α)、白细胞介素-6 (IL-6)和白细胞介素-10 (IL-10)水平,放射免疫法检测血清睾酮(T)和皮质酮(Cor)水平。结果与OM组相比,OOM组大鼠体重、脾脏指数及血清T/Cor比值均显著升高(P<0.05或P<0.01),脾脏单核细胞表面TLR4表达率显著降低(P<0.05),脾脏p38 MAPK、p-p38 MAPK和NF-κB蛋白质表达水平及p-p38/p38 MAPK比值均降低(P<0.05),脾脏TNF-α和IL-6水平显著降低(P<0.01),IL-10水平显著升高(P<0.01),脾脏Bcl-2/Bax比值升高(P<0.05),脾脏细胞凋亡水平降低(P<0.05),组织形态明显改善。结论 6 w递增负荷游泳训练期间补充OPCs可以通过调控TLR4/p38 MAPK/NF-κB信号通路相关蛋白质表达,有效缓解过度训练,抑制脾脏炎症因子过度表达及细胞凋亡,保护脾脏。[营养学报,2020,42 (5):479-484]
Objective To study the mechanism of spleen protection of oligomeric proanthocyanidins(OPCs) in overtraining rats by regulating the expression of toll-like receptor 4(TLR4)/p38 mitogen-activated protein kinase(p38 MAPK)/nuclear factor-kappa B(NF-κB) signaling pathway-related proteins.Methods Male Wistar rats were divided into control group(C group,n=12),overtraining group(OM group,n=12),OPCs+overtraining group(OOM group,n=13).C Group did not undergo any exercise intervention.OM and OOM group underwent 6-week incremental load swimming training.During the training,the OOM group was given 150 mg/(kg·d) OPCs in the volume of 5 ml/kg,and the other groups were given an equal volume of distilled water.twenty-four hours after the last training,the spleen index was calculated by weighing,splenic pathological changes were observed by HE staining,and splenic apoptosis level was detected by Tunel method.The expressions of splenic p38 MAPK,phosphorylated p38 mitogen-activated protein kinase(p-p38 MAPK),NF-κB,B cell lymphoma-2 protein(Bcl-2) and Bcl-2 associated X protein(Bax) were detected by immunohistochemical method.TLR4 expression rate on splenic monocytes surface was detected by flow cytometry.Splenic tumor necrosis factor-a(TNF-α),interleukin-6(IL-6) and interleukin-10(IL-10) levels were detected by enzyme-linked immunosorbent assay.Serum testosterone(T) and corticosterone(Cor) levels were detected by radioimmunoassay.Results Compared with the OM group,the body weight,spleen index and serum T/Cor ratio were increased(P<0.05 or P<0.01),TLR4 expression rate on splenic monocytes surface decreased(P<0.05),the expressions of splenic p38 MAPK,p-p38 MAPK,NF-κB and p-p38/p38 MAPK ratio decreased(P<0.05),splenic TNF-α and IL-6 level decreased(P<0.01),IL-10 level increased(P<0.01),splenic Bcl-2/Bax ratio increased(P<0.05),splenic apoptosis level reduced(P<0.05),and splenic histomorphological changes relieved in OOM group,.Conclusion OPCs supplementation during 6-week incremental load swimming training can regulate the expression of TLR4/p38 MAPK/NF-κB signaling pathway-related proteins,inhibit overexpression of splenic inflammatory cytokines and apoptosis,and thereby protect spleen effectively.[ACTA NUTRIMENTA SINICA,2020,42(5):479-484]
作者
胡戈
曹建民
秦菲
周海涛
郭娴
牛衍龙
霍海玉
HU Ge;CAO Jian-min;QIN Fei;ZHOU Hai-tao;GUO Xian;NIU Yan-long;HUO Hai-yu(Physical Education College,Changzhou University,Changzhou 213164;Beijing Sport University,Beijing 100084;Beijing Union University,Beijing 100023;Key Laboratory of Bioactive Substances and Functional Foods,Beijing Union University,Beijing 100191;Gannan Medical University,Ganzhou 341000,China)
出处
《营养学报》
CAS
CSCD
北大核心
2020年第5期479-484,共6页
Acta Nutrimenta Sinica
基金
北京市朝阳区协同创新项目(CYXC1817)
北京市高等学校高水平人才交叉培养“实培计划”项目
北京联合大学“启明星”大学生科技创新创业项目(No.201911417SJ159)
北京联合大学科研项目(No.ZK70202005)。
作者简介
胡戈(1982-),男,博士,讲师,E-mail:wizardhugo@163.com;通信作者:周海涛,E-mail:zsettle@sina.com。
