Attenuation of the Induction of TLRs 2 and 4 Mitigates Inflammation and Promotes Neurological Recovery After Focal Cerebral Ischemia.
Attenuation of the Induction of TLRs 2 and 4 Mitigates Inflammation and Promotes Neurological Recovery After Focal Cerebral Ischemia.
复制标题
DOI：
10.1007/s12975-020-00884-z
复制
发表时间：
2021-10
影响因子：
6.9
通讯作者：
Veeravalli KK
中科院分区：
文献类型：
--
作者：
Nalamolu KR;Challa SR;Fornal CA;Grudzien NA;Jorgenson LC;Choudry MM;Smith NJ;Palmer CJ;Pinson DM;Klopfenstein JD;Veeravalli KK
局灶性脑缺血后在大脑中引发的强烈炎症反应是有害的。最近，我们发现抑制Toll样受体(TLRs)2和4可以缩小缺血脑内的梗塞面积，减少促炎细胞因子的表达。在本研究中，我们检测了无限制地诱导TLR2和TLR4对再灌注一周后其下游信号分子和促炎细胞因子表达的影响。本研究的主要目的是研究同时敲除TLR2和TLR4对小胶质细胞M1/M2极化动力学和卒中后神经功能缺失及恢复的影响。采用单丝结扎大脑中动脉闭塞(MCAO)的方法建立幼年雄性SD大鼠短暂性局灶性脑缺血模型。再灌注后即刻经尾静脉注射TLR2shRNA和TLR4shRNA(T2sh+T4sh)纳米粒(T2sh和T4sh各1 mg/kg)或杂乱序列插入载体(载体对照)。不同队列的动物在再灌流期间被安乐死，分离缺血脑组织进行聚合酶链式反应，然后进行琼脂糖凝胶电泳、实时定量聚合酶链式反应、免疫印迹和免疫荧光分析。适当的组定期接受一系列标准的神经学测试，直到再灌流后14天。TLR2和TLR4及其下游信号分子包括促炎细胞因子的表达在再灌流后1周仍有增加。再灌注后即刻T2sh+T4sh治疗可减轻缺血后炎症反应，保护运动功能，促进感觉和运动功能的恢复。我们的结论是，缺血后TLR2和TLR4的诱导在再灌注后至少持续七天，有助于急性炎症的严重程度，并阻碍神经恢复。与先前在TLR 2或4基因敲除模型中的研究不同，这项研究在药物相关的临床前啮齿动物卒中模型中的结果具有翻译意义。
The intense inflammatory response triggered in the brain after focal cerebral ischemia is detrimental. Recently we showed that the suppression of toll-like receptors (TLRs) 2 and 4 attenuates infarct size and reduces the expression of pro-inflammatory cytokines in the ischemic brain. In this study, we tested the effect of unrestricted induction of TLRs 2 and 4 on the expression of its downstream signaling molecules and pro-inflammatory cytokines one week after reperfusion. The primary purpose of this study was to investigate the effect of simultaneous knockdown of TLRs 2 and 4 on M1/M2 microglial polarization dynamics and post-stroke neurological deficits and the recovery. Transient focal cerebral ischemia was induced in young adult male Sprague-Dawley rats by the middle cerebral artery occlusion (MCAO) procedure using a monofilament suture. Appropriate cohorts of rats were treated with a nanoparticle formulation of TLR2shRNA and TLR4shRNA (T2sh+T4sh) expressing plasmids (1 mg/kg each of T2sh and T4sh) or scrambled sequence inserted vector (vehicle control) expressing plasmids (2 mg/kg) intravenously via tail vein immediately after reperfusion. Animals from various cohorts were euthanized during reperfusion, and the ischemic brain tissue was isolated and utilized for PCR followed by agarose gel electrophoresis, real time PCR, immunoblot, and immunofluorescence analysis. Appropriate groups were subjected to a battery of standard neurological tests at regular intervals until fourteen days after reperfusion. The increased expression of both TLRs 2 and 4 and their downstream signaling molecules including the pro-inflammatory cytokines was observed even at one-week after reperfusion. T2sh+T4sh treatment immediately after reperfusion attenuated the post-ischemic inflammation, preserved the motor function, and promoted recovery of the sensory and motor functions. We conclude that the post-ischemic induction of TLRs 2 and 4 persists for at least seven days after reperfusion, contributes to the severity of acute inflammation, and impedes neurological recovery. Unlike previous studies in TLRs 2 or 4 knockout models, results of this study in a pharmacologically relevant preclinical rodent stroke model have translational significance.
登录
查看更多内容
影响因子：
3.4
作者：
Jin, Rong;Liu, Lin;Zhang, Shihao;Nanda, Anil;Li, Guohong
通讯作者：
Li, Guohong
影响因子：
6.3
作者：
Brea, David;Blanco, Miguel;Castillo, Jose
通讯作者：
Castillo, Jose
影响因子：
6.2
作者：
Durafourt, Bryce A.;Moore, Craig S.;Antel, Jack P.
通讯作者：
Antel, Jack P.
影响因子：
--
作者：
Chelluboina, Bharath;Nalamolu, Koteswara Rao;Veeravalli, Krishna Kumar
通讯作者：
Veeravalli, Krishna Kumar
影响因子：
3.7
作者：
Andresen L;Theodorou K;Grünewald S;Czech-Zechmeister B;Könnecke B;Lühder F;Trendelenburg G
通讯作者：
Trendelenburg G