丹参酮ⅡA通过抑制脊髓HMGB1表达缓解炎性痛大鼠痛觉过敏的研究
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摘要
目的:探讨丹参酮ⅡA (TSN ⅡA)对完全弗氏佐剂(CFA)引起的炎性痛大鼠机械性痛敏的影响及其作用机制。方法:雄性Wistar大鼠完全随机数字法分为4组(n=6只):正常对照组(Control)、TSA ⅡA处理组(TSA ⅡA)、炎性痛组(CFA)、炎性痛+TSA ⅡA处理组(CFA+TSA ⅡA)。利用CFA注射大鼠左下肢足底制作炎性痛大鼠模型,各组大鼠分别用腹腔注射法给予20 mg/kg TSN ⅡA或等量生理盐水,利用von Frey丝检测各组大鼠的痛行为学变化,利用Western Blot检测各组大鼠脊髓Toll样受体4(TLR4)和高迁移率蛋白-1(HMGB1)蛋白表达变化,利用Real-time RT-PCR检测TNF-α、IL-1β和IL-6等分子m RNA表达变化。结果:CFA可诱发大鼠出现明显的痛敏反应,TSN ⅡA可以显著地缓解炎性痛大鼠机械性痛敏; CFA引起的炎性痛大鼠脊髓TLR4、HMGB1、TNF-α、IL-1β和IL-6等分子表达增加,而TSN ⅡA可以显著逆转上述分子的表达上调。结论:TSN ⅡA可通过抑制HMGB1-TLR4通路缓解CFA引起的大鼠炎性痛。
Objective: To observe the effect of tanshinone ⅡA(tanshinone ⅡA,TSN ⅡA) on mechanical allodynia in the inflammatory pain rats induced by complete Freund's adjuvant(complete Freund's adjuvant,CFA) and to explore the mechanisms. Methods: Wistar male rats were randomly divided into 4 groups(n = 6) : control group,TSA ⅡA group,CFA group,and CFA + TSA ⅡA group. CFA was injected into the ratsl'eft paws to make a rat model of inflammatory pain,and the behavioral changes of rats were observed by von Frey fibers. The Toll-like receptors 4(TLR4)and high mobility protein-1(HMGB1) protein expression of the spinal cord of each group were detected by Western Blot and the expression change of TNF-α,IL-1β and IL-6 were detected by Real-time RT-PCR. Results: CFA can induce the obvious pain allodynia in rats. Intraperitoneal injection of TSN ⅡA could reverse the mechanical allodynia in inflammatory pain rats. The expression of TLR4,HMGB1,and other inflammatory cytokines in the spinal cord of inflammatory pain rats were significantly increased. TSN ⅡA could significantly reduce the expression of TLR4,HMGB1,and proinflammatory cytokines. Conclusion: TSN ⅡA could play an analgesic role on inflammatory pain rats through inhibiting the HMGB1-TLR4 pathway.
Objective: To observe the effect of tanshinone ⅡA(tanshinone ⅡA,TSN ⅡA) on mechanical allodynia in the inflammatory pain rats induced by complete Freund's adjuvant(complete Freund's adjuvant,CFA) and to explore the mechanisms. Methods: Wistar male rats were randomly divided into 4 groups(n = 6) : control group,TSA ⅡA group,CFA group,and CFA + TSA ⅡA group. CFA was injected into the ratsl'eft paws to make a rat model of inflammatory pain,and the behavioral changes of rats were observed by von Frey fibers. The Toll-like receptors 4(TLR4)and high mobility protein-1(HMGB1) protein expression of the spinal cord of each group were detected by Western Blot and the expression change of TNF-α,IL-1β and IL-6 were detected by Real-time RT-PCR. Results: CFA can induce the obvious pain allodynia in rats. Intraperitoneal injection of TSN ⅡA could reverse the mechanical allodynia in inflammatory pain rats. The expression of TLR4,HMGB1,and other inflammatory cytokines in the spinal cord of inflammatory pain rats were significantly increased. TSN ⅡA could significantly reduce the expression of TLR4,HMGB1,and proinflammatory cytokines. Conclusion: TSN ⅡA could play an analgesic role on inflammatory pain rats through inhibiting the HMGB1-TLR4 pathway.
引文
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[2]Andersson U,Erlandsson-Harris H,Yang H,et al.HMGB1 as a DNA-binding cytokine[J].J Leukoc Biol,2002,72(6):1084-1091.
[3]Yamada S,Maruyama I.HMGB1,a novel inflammatory cytokine[J].Clin Chim Acta,2007,375(1):36-42.DOI:10.1016/j.cca.2006.07.019.
[4]Zhang Y,Mou J,Cao L,et al.Micro RNA-142-3p relieves neuropathic pain by targeting high mobility group box 1[J].Int J Mol Med,2018,41(1):501-510.DOI:10.3892/ijmm.2017.3222.
[5]Tong W,Wang W,Huang J,et al.Spinal high-mobility group box 1contributes to mechanical allodynia in a rat model of bone cancer pain[J].Biochem Biophys Res Commun,2010,395(4):572-576.DOI:10.1016/j.bbrc.2010.04.086.
[6]Matsuura W,Harada S,Liu K,et al.Evidence of a role for spinal HMGB1 in ischemic stress-induced mechanical allodynia in mice[J].Brain Res,2018,1687:1-10.DOI:10.1016/j.brainres.2018.02.026.
[7]Agalave NM,Larsson M,Abdelmoaty S,et al.Spinal HMGB1 induces TLR4-mediated long-lasting hypersensitivity and glial activation and regulates pain-like behavior in experimental arthritis[J].Pain,2014,155(9):1802-1813.DOI:10.1016/j.pain.2014.06.007.
[8]Zhang GX,Zhang YY,Zhang XX,et al.Different network pharmacology mechanisms of Danshen-based Fangjis in the treatment of stable angina[J].Acta Pharmacol Sin.2018,39(6):952-960.DOI:10.1038/aps.2017.191.
[9]Li QN,Huang ZP,Gu QL,et al.Synthesis and biological evaluation of novel tanshinone IIA derivatives for treating pain[J].Chin J Nat Med.2018,16(2):113-124.DOI:10.1016/S1875-5364(18)30037-2.
[10]Hao W,Chen L,Wu LF,et al.Tanshinone IIA exerts an antinociceptive effect in rats with cancer-induced bone pain[J].Pain Physician.2016,19(7):465-476.
[11]汪伟,王文,武胜昔,等.炎症性痛和神经病理性痛模型[J].神经解剖学杂志,2007,23(1):93-98.
[12]Zhao JY,Duan XL,Yang L,et al.Activity-dependent synaptic recruitment of Neuroligin 1 in spinal dorsal horn contributed to inflammatory pain[J].Neuroscience.2018,388:1-10.DOI:10.1016/j.neuroscience.2018.06.047.
[13]Sun L,Li M,Ma X,et al.Inhibition of HMGB1 reduces rat spinal cord astrocytic swelling and AQP4 expression after oxygen-glucose deprivation and reoxygenation via TLR4 and NF-kappaB signaling in an IL-6-dependent manner[J].J Neuroinflammation,2017,14(1):231.DOI:10.1186/s12974-017-1008-1.
[14]Wang X,Feng C,Qiao Y,et al.Sigma 1 receptor mediated HMGB1 expression in spinal cord is involved in the development of diabetic neuropathic pain[J].Neurosci Lett,2018,668:164-168.DOI:10.1016/j.neulet.2018.02.002.
[15]Wang YS,Li YY,Wang LH,et al.Tanshinone IIA attenuates chronic pancreatitis-induced pain in rats via downregulation of HMGB1 and TRL4 expression in the spinal cord[J].Pain Physician,2015,18(4):E615-628.
[16]Kuang X,Huang Y,Gu HF,et al.Effects of intrathecal epigallocatechin gallate,an inhibitor of Toll-like receptor 4,on chronic neuropathic pain in rats[J].Eur J Pharmacol,2012,676(1-3):51-56.DOI:10.1016/j.ejphar.2011.11.037.
[17]Chen XL,Sun L,Guo F,et al.High-mobility group box-1 induces proinflammatory cytokines production of Kupffer cells through TLRsdependent signaling pathway after burn injury[J].PLOS ONE,2012,7(11):e50668.DOI:10.1371/journal.pone.0050668.
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[19]Liu T,Q Han,G Chen,et al.Toll-like receptor 4 contributes to chronic itch,alloknesis,and spinal astrocyte activation in male mice[J].Pain,2016,157(4):806-817.DOI:10.1097/j.pain.0000000000000439.
[20]Hu Y,Z Wang,S Pan,et al.Melatonin protects against bloodbrain barrier damage by inhibiting the TLR4/NF-kappaB signaling pathway after LPS treatment in neonatal rats[J].Oncotarget,2017,8(19):31638-31654.DOI:10.18632/oncotarget.15780.
[21]Bruno K,Woller S,Miller Y,et al.Targeting toll-like receptor-4(TLR4)-an emerging therapeutic target for persistent pain states[J].Pain,2018,159(10):1908-1915.DOI:10.1097/j.pain.0000000000001306.
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