基于LTB4/BLTR通路探讨黄芩苷-栀子苷配伍对局灶性脑缺血大鼠再灌注损伤的影响  

Effect of baicalin-geniposide compatibility on the brain tissue injury in rats with focal cerebral ischemia-reperfusion via LTB4/BLTR pathway

作　　者：周爽 李慧敏 张行行 史永恒[1] 王川[1] 王国全[1] 李敏[1] 王斌[1] ZHOU Shuang;LI Hui-min;ZHANG Hang-hang;SHI Yong-heng;WANG Chuan;WANG Guo-quan;LI Min;WANG Bin(Shaanxi University of Traditional Chinese Medicine,Xianyang Shaanxi 712046)

机构地区：[1]陕西中医药大学,陕西咸阳712046

出　　处：《中南药学》2022年第6期1255-1261,共7页Central South Pharmacy

基　　金：国家自然科学基金项目(No.81473385);陕西省中医药优秀中青年科技骨干人才队伍建设项目(序号:2);2020年度陕西中医药大学创新创业项目(No.2020CX36)。

摘　　要：目的基于LTB4/BLTR通路探讨预防及治疗脑损伤的分子机制,探究黄芩苷-栀子苷治疗脑缺血的抗炎作用机制。方法将SD雄性大鼠随机分为假手术组,模型组,白三烯B4受体(BLTR)抑制剂组(LY255283,1 mg·kg^(-1)),黄芩苷-栀子苷7∶3配伍组低、高剂量组(BC/GP组,45 mg·kg^(-1)、60 mg·kg^(-1))和BC/GP联合抑制剂组(BC/GP45+LY255283、BC/GP60+LY255283)。线栓法构建大鼠脑中动脉阻塞模型,模拟局灶性脑缺血再灌注。缺血2 h后拔出线栓实现再灌注,缺血24 h后取材。在2 h和24 h分别进行Zea-Longa评分和神经缺损程度评分(mNSS)。HE染色观察组织病理形态的变化;酶联免疫法检测脑组织中白三烯A4水解酶(LTA4H)和白三烯B4(LTB4)、髓过氧化物酶(MPO)的表达,以及炎性因子IL-1β和促炎因子TNF-α的表达;qPCR检测BLT1、BLT2 mRNA表达。结果与假手术组相比,模型组大鼠的神经行为学评分显著增加(P<0.01),脑组织中LTA4、LTB4、MPO、BLT1、BLT2、TNF-α和IL-1β的表达均显著增加。与模型组比,给予BC/GP后mNSS评分显著降低(P<0.01),缺血区炎性细胞浸润及组织水肿减轻,TNF-α、IL-1β、LTB4、LTA4H、MPO、BLT1和BLT2 mRNA表达显著降低(P<0.01)。结论黄芩苷-栀子苷(7∶3)配伍可以有效改善脑缺血再灌注损伤,其作用机制可能与下调LTB4/BLTR通路有关。Objective To determine the molecular mechanism of preventing and treating brain injury based on LTB4/BLTR pathway,and the anti-inflammation mechanism of baicalin-gardenoside for cerebral ischemia.Methods Male SD rats were randomly divided into a sham group,a model group,a leukotriene B4 receptor(BLTR)inhibitor group(LY255283,1 mg·kg^(-1)),baicalin gardenoside compatibility(7∶3)low and high dose groups(BC/GP,45 mg·kg^(-1) and 60 mg·kg^(-1))and a BC/GP combined inhibitor group(BC/GP45+LY255283;BC/GP60+LY255283).The rat model of middle cerebral artery occlusion(MACO)was established by thread occlusion to simulate focal cerebral ischemia-reperfusion.After 2 hours of ischemia,the lead bolt was pulled out for the reperfusion,and the samples were taken after 24 hours of ischemia.Zea-longa score and nerve defect degree score(mNSS)were taken at 2 h and 24 h,respectively.The histopathological changes were observed by HE staining.The expressions of leukotriene A4 hydrolase(LTA4H),leukotriene B4(LTB4)and myeloperoxide(MPO)in the brain tissue and the expression of inflammatory factor IL-1βand proinflammatory factor TNF-αwere detected by enzyme-linked immunosorbent assay.The expression of BLT1 and BLT2 mRNA was detected by qPCR.Results Compared with the sham group,the neurobehavioral score of the model group was significantly increased(P<0.01),and the expressions of LTA4,LTB4,MPO,BLT1,BLT2,TNF-αand IL-1βwere significantly increased.Compared with the model group,the mNSS score was decreased significantly after BC/GP treatment(P<0.01),the inflammatory cell infiltration and tissue edema in the ischemic area were alleviated.The expressions of TNF-α,IL-1β,LTB4,LTA4H,MPO,BLT1 and BLT2 mRNA were significantly inhibited(P<0.01).Conclusion Baicalin-geniposide(7∶3)combination can effectively improve the cerebral ischemia-reperfusion injury,whose mechanism is related to the down regulaton of LTB4/BLTR pathway.

关 键 词：脑缺血再灌注损伤 白三烯B4 黄芩苷 栀子苷 白三烯B4受体 

分 类 号：R284[医药卫生—中药学]