红景天苷抑制补体成分对脑缺血/再灌注大鼠的神经保护作用
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摘要
目的研究红景天苷对脑缺血/再灌注大鼠补体成分的作用,进一步探讨红景天苷神经保护的作用机制。方法♂成年SD大鼠45只,随机分为假手术(Sham)组、模型(MCAO)组、红景天苷(MCAO+Sal)组,线栓法制备大鼠大脑中动脉闭塞模型(MCAO),红景天苷浓度为50 mg·kg-1·d-1,连续给药6 d。检测脑组织线粒体内Bax、Bcl-xl蛋白表达,基因芯片聚类分析,q PCR检测补体成分C1qa、C1qb、C1qc等mRNA表达,Western blot检测补体C3的表达,免疫荧光法检测C3和Neu N的表达。结果与MCAO组相比,红景天苷给药后,降低线粒体内Bax蛋白表达,促进Bcl-xl蛋白表达;经基因芯片聚类分析及q PCR检测,红景天苷能够抑制C1qa、C1qb、C1qc等mRNA补体相关基因表达;经Western blot及免疫荧光法检测,红景天苷能够抑制C3的表达,同时促进Neu N的表达。结论红景天苷能够抑制补体成分,尤其是补体中枢成分C3,对MCAO大鼠具有神经保护作用。
Aim To study the effects of salidroside on the complement components in MCAO rats and to further explore the mechanism of neuroprotection of salidroside. Methods Forty-five healthy male SpragueDawley rats were randomly divided into sham operation( Sham) group,model( MCAO) group,and salidroside( MCAO + Sal) group. Rats were administered salidroside,or vehicle,daily for 6 days,after middle cerebral artery occlusion( MCAO) 2 h and reperfusion 1 h.The protein expressions of C3 and brain tissue mitochondrial Bax,Bcl-xl were detected by Western blot.The mRNA expressions of C1 qa,C1 qb,C1 qc,C2,C3,C3 a, C4 a, C5 a, Cfh, Cfd were detected by qPCR. Gene chip technology was used for detection.The expressions of C3 and NeuN were detected by immunofluorescence. Results Compared with MCAOgroup,salidroside could reduce the protein expression of Bax protein in mitochondria and promote the expression of Bcl-xL protein after salidroside treatment. By gene chip cluster analysis and qPCR detection,salidroside could inhibit the expression of complement-related genes such as C1 qa,C1 qb,C1 qc,C2,C3,C3 a,C4 a,C5 ar,Cfh and Cfd mRNA. According to Western blot and immunofluorescence,salidroside inhibited the expression of C3 and promoted the expression of NeuN. Conclusions Salidroside can inhibit the complement component,especially the action of the complement central component C3,exerting neuroprotective effects on MCAO rats.
Aim To study the effects of salidroside on the complement components in MCAO rats and to further explore the mechanism of neuroprotection of salidroside. Methods Forty-five healthy male SpragueDawley rats were randomly divided into sham operation( Sham) group,model( MCAO) group,and salidroside( MCAO + Sal) group. Rats were administered salidroside,or vehicle,daily for 6 days,after middle cerebral artery occlusion( MCAO) 2 h and reperfusion 1 h.The protein expressions of C3 and brain tissue mitochondrial Bax,Bcl-xl were detected by Western blot.The mRNA expressions of C1 qa,C1 qb,C1 qc,C2,C3,C3 a, C4 a, C5 a, Cfh, Cfd were detected by qPCR. Gene chip technology was used for detection.The expressions of C3 and NeuN were detected by immunofluorescence. Results Compared with MCAOgroup,salidroside could reduce the protein expression of Bax protein in mitochondria and promote the expression of Bcl-xL protein after salidroside treatment. By gene chip cluster analysis and qPCR detection,salidroside could inhibit the expression of complement-related genes such as C1 qa,C1 qb,C1 qc,C2,C3,C3 a,C4 a,C5 ar,Cfh and Cfd mRNA. According to Western blot and immunofluorescence,salidroside inhibited the expression of C3 and promoted the expression of NeuN. Conclusions Salidroside can inhibit the complement component,especially the action of the complement central component C3,exerting neuroprotective effects on MCAO rats.
引文
[1] Chen S,Chen Z,Cui J,et al. Early abrogation of gelatinase activity extends the time window for t PA thrombolysis after embolic focal cerebral ischemia in mice[J]. e Neuro,2018,5(3):0391-17.
[2]赖文芳,张小琴,洪海棉,等.红景天苷对大鼠局灶性脑缺血/再灌注损伤的神经保护作用[J].中国药理学通报,2015,31(6):775-80.[2] Lai W F,Zhang X Q,Hong H M,et al. Neuroprotective effects of salidroside against focal celebral ischemia/reperfusion injury in rats[J]. Chin Pharmacol Bull,2015,31(6):775-80.
[3]谢秀利,洪海棉,赖文芳,等.红景天苷对大脑中动脉闭塞模型大鼠的神经保护作用[J].中国药理学通报,2015,31(10):1452-7.[3] Xie X L,Hong H M,Lai W F,et al. Protective effects of salidroside in MCAO rats[J]. Chin Pharmacol Bull,2015,31(10):1452-7.
[4]宋百颖,赖文芳,苏燕青,等.红景天苷通过抑制补体C3的表达对MCAO大鼠缺血侧NeuN、Egr4影响的研究[J].中国药理学通报,2017,33(11):1579-84.[4] Song B Y,Lai W F,Su Y Q,et al. Effects of salidroside on expression of NeuN and Egr4 in ischemic side of MCAO rats by inhibiting expression of C3[J]. Chin Pharmacol Bull,2017,33(11):1579-84.
[5] Lai W,Xie X,Zhang X,et al. Inhibition of complement drives increase in early growth response proteins and neuroprotection mediated by salidroside after cerebral ischemia[J]. Inflammation,2017,41(2):1-15.
[6] He S,Atkinson C,Evans Z,et al. A role for complement in the enhanced susceptibility of steatotic livers to ischemia and reperfusion injury[J]. J Immunol,2009,183(7):4764-72.
[7]鲜尽红,何莉,汪正清.补体级联反应在脑缺血/再灌注炎症损伤中的作用[J].创伤外科杂志,2011,13(6):561-3.[7] Xian J H,He L,Wang Z Q. Role of complement cascade in cerebral ischemia/reperfusion inflammatory injury[J]. J Traumatic Surgery,2011,13(6):561-3.
[8] Lai W,Zheng Z,Zhang X,et al. Salidroside-mediated neuroprotection is associated with induction of early growth response genes(Egrs)across a wide therapeutic window[J]. Neurotox Res,2015,28(2):108-21.
[9] Széplaki G,Szegedi R,Hirschberg K,et al. Strong complement activation after acute ischemic stroke is associated with unfavorable outcomes[J]. Atherosclerosis,2009,204(1):315-20.
[10] Peng B,Zhao P,Lu Y P,et al. Z-ligustilide activates the Nrf2/HO-1 pathway and protects against cerebral ischemia-reperfusion injury in vivo and in vitro[J]. Brain Res,2013,1520(32):168-77.
[11] Goldstein L B. Acute ischemic stroke treatment in 2007[J]. Circulation,2007,116(13):1504-14.
[12] Lucas S M,Rothwell N J,Gibson R M. The role of inflammation in CNS injury and disease[J]. Br J Pharmacol,2010,147(S1):S232-40.
[13] Morgan B P,Gasque P. Extrahepatic complement biosynthesis:where,when and why[J]? Clin Exp Immunol,2010,107(1):1-7.
[2]赖文芳,张小琴,洪海棉,等.红景天苷对大鼠局灶性脑缺血/再灌注损伤的神经保护作用[J].中国药理学通报,2015,31(6):775-80.[2] Lai W F,Zhang X Q,Hong H M,et al. Neuroprotective effects of salidroside against focal celebral ischemia/reperfusion injury in rats[J]. Chin Pharmacol Bull,2015,31(6):775-80.
[3]谢秀利,洪海棉,赖文芳,等.红景天苷对大脑中动脉闭塞模型大鼠的神经保护作用[J].中国药理学通报,2015,31(10):1452-7.[3] Xie X L,Hong H M,Lai W F,et al. Protective effects of salidroside in MCAO rats[J]. Chin Pharmacol Bull,2015,31(10):1452-7.
[4]宋百颖,赖文芳,苏燕青,等.红景天苷通过抑制补体C3的表达对MCAO大鼠缺血侧NeuN、Egr4影响的研究[J].中国药理学通报,2017,33(11):1579-84.[4] Song B Y,Lai W F,Su Y Q,et al. Effects of salidroside on expression of NeuN and Egr4 in ischemic side of MCAO rats by inhibiting expression of C3[J]. Chin Pharmacol Bull,2017,33(11):1579-84.
[5] Lai W,Xie X,Zhang X,et al. Inhibition of complement drives increase in early growth response proteins and neuroprotection mediated by salidroside after cerebral ischemia[J]. Inflammation,2017,41(2):1-15.
[6] He S,Atkinson C,Evans Z,et al. A role for complement in the enhanced susceptibility of steatotic livers to ischemia and reperfusion injury[J]. J Immunol,2009,183(7):4764-72.
[7]鲜尽红,何莉,汪正清.补体级联反应在脑缺血/再灌注炎症损伤中的作用[J].创伤外科杂志,2011,13(6):561-3.[7] Xian J H,He L,Wang Z Q. Role of complement cascade in cerebral ischemia/reperfusion inflammatory injury[J]. J Traumatic Surgery,2011,13(6):561-3.
[8] Lai W,Zheng Z,Zhang X,et al. Salidroside-mediated neuroprotection is associated with induction of early growth response genes(Egrs)across a wide therapeutic window[J]. Neurotox Res,2015,28(2):108-21.
[9] Széplaki G,Szegedi R,Hirschberg K,et al. Strong complement activation after acute ischemic stroke is associated with unfavorable outcomes[J]. Atherosclerosis,2009,204(1):315-20.
[10] Peng B,Zhao P,Lu Y P,et al. Z-ligustilide activates the Nrf2/HO-1 pathway and protects against cerebral ischemia-reperfusion injury in vivo and in vitro[J]. Brain Res,2013,1520(32):168-77.
[11] Goldstein L B. Acute ischemic stroke treatment in 2007[J]. Circulation,2007,116(13):1504-14.
[12] Lucas S M,Rothwell N J,Gibson R M. The role of inflammation in CNS injury and disease[J]. Br J Pharmacol,2010,147(S1):S232-40.
[13] Morgan B P,Gasque P. Extrahepatic complement biosynthesis:where,when and why[J]? Clin Exp Immunol,2010,107(1):1-7.