Inhibition of RhoA/Rho kinase signaling pathway by fasudil protects against kainic acid-induced neurite injury.
Inhibition of RhoA/Rho kinase signaling pathway by fasudil protects against kainic acid-induced neurite injury.
复制标题
Fasudil抑制RhoA/Rho激酶信号传导途径可预防海藻酸诱导的神经突损伤。
DOI：
10.1002/brb3.2266
复制
发表时间：
2021-08
影响因子：
3.1
通讯作者：
Zeng LH
中科院分区：
文献类型：
--
作者：
Xiang Y;Niu Y;Xie Y;Chen S;Zhu F;Shen W;Zeng LH
RhoA/Rho 激酶途径对于调节细胞骨架结构至关重要。尽管它对正常神经突生长的影响已被证明，但该途径在癫痫引起的神经突损伤中的作用尚未揭示。该研究检测了 RhoA/Rho 激酶信号通路的磷酸化水平，并阐明法舒地尔对红藻氨酸 (KA) 处理的 Neuro-2A 细胞和海马神经元中 RhoA/Rho 激酶信号通路和神经突生长的影响。采用Western blotting分析RhoA/Rho激酶信号通路关键蛋白的表达及肌动蛋白的解聚情况。无血清孵育诱导神经突生长后，固定 Neuro-2A 细胞，并应用罗丹明鬼笔环肽免疫荧光法检测细胞形态和神经突长度。在原代海马神经元中检测了KA对神经元的影响。在培养的神经元或海马切片中进行全细胞膜片钳以记录动作电位。 100-200 μmol/L剂量的KA诱导Rho相关卷曲螺旋蛋白激酶磷酸化增加，Lin11、Isl-1和Mec-3激酶和cofilin磷酸化减少。 200 μmol/L KA的作用在1~2小时达到峰值，8小时后逐渐恢复到基线。 Rho 激酶抑制剂法舒地尔预处理逆转了 KA 诱导的 RhoA/Rho 激酶通路激活，并增加了 slingshot 和 14-3-3 的磷酸化，从而降低了 G/F-肌动蛋白的比率。 KA 治疗可诱导 Neuro-2a 细胞和培养的海马神经元中神经突生长的抑制和棘的减少，而法舒地尔预处理可减轻 KA 诱导的神经突生长抑制和棘的损失。这些数据表明，抑制 RhoA/Rho 激酶通路可能是癫痫引起的损伤的潜在治疗方法。 100～200μmol/L红藻氨酸诱导ROCK磷酸化增加，LIMK和cofilin磷酸化减少。 Rho 激酶抑制剂 Fasudil 预处理可逆转 KA 诱导的 RhoA/Rho 激酶通路激活以及 SSH 和 14-3-3 磷酸化的增加。 Fasudil 预处理减轻了 KA 诱导的神经突生长抑制和脊柱损失。
RhoA/Rho kinase pathway is essential for regulating cytoskeletal structure. Although its effect on normal neurite outgrowth has been demonstrated, the role of this pathway in seizure‐induced neurite injury has not been revealed. The research examined the phosphorylation level of RhoA/Rho kinase signaling pathway and to clarify the effect of fasudil on RhoA/Rho kinase signaling pathway and neurite outgrowth in kainic acid (KA)‐treated Neuro‐2A cells and hippocampal neurons. Western blotting analysis was used to investigate the expression of key proteins of RhoA/Rho kinase signaling pathway and the depolymerization of actin. After incubated without serum to induce neurite outgrowth, Neuro‐2A cells were fixed, and immunofluorescent assay of rhodamine‐phalloidin was applied to detect the cellular morphology and neurite length. The influence of KA on neurons was detected in primary hippocampal neurons. Whole‐cell patch clamp was conducted in cultured neurons or hippocampal slices to record action potentials. KA at the dose of 100–200 μmol/L induced the increase in phosphorylation of Rho‐associated coiled‐coil‐containing protein kinase and decrease in phosphorylation of Lin11, Isl‐1 and Mec‐3 kinase and cofilin. The effect of 200 μmol/L KA was peaked at 1–2 hours, and then gradually returned to baseline after 8 hours. Pretreatment with Rho kinase inhibitor fasudil reversed KA‐induced activation of RhoA/Rho kinase pathway and increase in phosphorylation of slingshot and 14‐3‐3, which consequently reduced the ratio of G/F‐actin. KA treatment induced inhibition of neurite outgrowth and decrease in spines both in Neuro‐2a cells and in cultured hippocampal neurons, and pretreatment with fasudil alleviated KA‐induced neurite outgrowth inhibition and spine loss. These data indicate that inhibiting RhoA/Rho kinase pathway might be a potential treatment for seizure‐induced injury. Kainic acid at the dose of 100 to 200 μmol/L induced the increase in phosphorylation of ROCK and decrease in phosphorylation of LIMK and cofilin. Pretreatment with Rho kinase inhibitor Fasudil reversed KA‐induced activation of RhoA/Rho kinase pathway and increase in phosphorylation of SSH and 14‐3‐3. Pretreatment with Fasudil alleviated KA‐induced neurite outgrowth inhibition and spine loss.
登录
查看更多内容
影响因子：
3.7
作者：
Salto R;Vílchez JD;Girón MD;Cabrera E;Campos N;Manzano M;Rueda R;López-Pedrosa JM
通讯作者：
López-Pedrosa JM
影响因子：
4.8
作者：
Tönges L;Koch JC;Bähr M;Lingor P
通讯作者：
Lingor P
影响因子：
7.3
作者：
Feng, Yangbo;LoGrasso, Philip V.;Li, Rongshi
通讯作者：
Li, Rongshi
影响因子：
4.7
作者：
Munnamalai V;Weaver CJ;Weisheit CE;Venkatraman P;Agim ZS;Quinn MT;Suter DM
通讯作者：
Suter DM
影响因子：
2
作者：
Dias, Luis Augusto;de Angelis, Geisa;Casulari, Luiz Augusto
通讯作者：
Casulari, Luiz Augusto