Rho kinase II interference by small hairpin RNA ameliorates 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced parkinsonism in mice

Abstract

Novel therapeutic targets are required for the treatment of Parkinson's disease (PD). Previous studies suggest that the Rho/Rho-associated, coiled-coil-containing protein kinases (ROCKs) signaling pathway may be a promising therapeutic target in PD. To elucidate the importance of ROCKII in the pathogenesis of dopaminergic (DA) neuron loss and to investigate the efficacy of ROCK inhibitors in PD, ROCKII expression in the substantia nigra (SN) of mice was silenced through the injection of a lentivirus-based small hairpin RNA system. Empty lentivirus vectors served as controls. Mice were subsequently challenged with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). The expression levels and activity of ROCKII were elevated in tyrosine hydroxylase-positive neurons and in cluster of differentiation (CD) 11b-positive microglia within the SN of MPTP-treated mice, which was accompanied by an increased level of expression of inducible nitric oxide synthase (iNOS) and activation of the Toll-like receptor (TLR)2/nuclear factor (NF)-κB signaling pathway in M1 microglia. ROCKII interference (RI) significantly improved movement disorder and attenuated DA neuron loss induced by MPTP. In addition, RI inhibited the activation of M1 microglia in the SN, exhibiting reduced activity of the TLR2/NF-κB signaling pathway and decreased expression levels of iNOS and inflammatory factors, including interleukin (IL)-1β and IL-6. The results of the present study verify that ROCKII participates in the loss of DA neurons induced by MPTP and suggest that ROCKII inhibition may be a promising therapeutic target for PD.

Nuclear factor‐kappa B (NF‐κB) in pathophysiology of Parkinson disease: Diverse patterns and mechanisms contributing to neurodegeneration

“…Several synthetic and natural substances, as summarized in Table 1, are utilized to attenuate dopaminergic neuron loss through suppressing TLR signaling pathway and NF-κB (Café-Mendes et al, 2017;Cheng & Zhu, 2019;Li et al, 2018;Ling et al, 2019;Lv et al, 2019;Xu et al, 2017;Yang et al, 2019;Yu et al, 2016;Zhang & Xu, 2018 ;Zhang et al, 2016 It has been demonstrated that aggregated α-synuclein by binding to CD36 on microglia, which by recruiting Fyn kinase can activate protein kinase C-delta (PKC-δ), thereby propagating translocation of NF-κB p65 subunit to the nucleus. In the nucleus, p65 subunit enhances expression of IL-1β and NLR family pyrin domain containing 3 (NLRP3) inflammasome (Panicker et al, 2019) (Figure 1).…”

“…Several synthetic and natural substances, as summarized in Table 1, are utilized to attenuate dopaminergic neuron loss through suppressing TLR signaling pathway and NF‐κB (Café‐Mendes et al., 2017; Cheng & Zhu, 2019; Li et al., 2018; Ling et al., 2019; Lv et al., 2019; Xu et al., 2017; Yang et al., 2019; Yu et al., 2016; Zhang & Xu, 2018 ​; Zhang et al., 2016).…”

NADPH-Oxidase, Rho-Kinase and Autophagy Mediate the (Pro)renin-Induced Pro-Inflammatory Microglial Response and Enhancement of Dopaminergic Neuron Death

“…ROCK activation has been shown to play a major role in the microglial pro-inflammatory response, including migration of microglia into the inflamed areas [ 24 , 51 , 52 , 53 ]. Our research group and others have shown that ROCK inhibitors have neuroprotective effects against dopaminergic degeneration in PD models [ 42 , 54 , 55 ], and that inhibition of the microglial pro-inflammatory response plays a major role in the neuroprotective effect [ 23 , 42 , 56 , 57 ]. More recently, we showed that the ROCK inhibitor fasudil inhibits the dyskinetic behavior induced by L-DOPA in PD models, which is mediated by inhibition of the neuroinflammatory response [ 29 ].…”

Recent developments in nucleic acid-based therapies for Parkinson’s disease: Current status, clinical potential, and future strategies

“…Although one of the published studies has reported that the shRNAs decreased the expression of α-synuclein and attenuated dopamine neuron degeneration in rat PD model induced with 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine (MPTP) ( Liu et al, 2014 ; Zhang et al, 2016 ), however, other techniques, such as decreasing inflammation via class II transactivator or caspase-1 inhibition by shRNA, have also been tried instead of directly targeting SNCA ( Li et al, 2020 ). In addition, shRNAs targeting nuclear receptor related 1 (Nurr1) or Src homology region 2 (SH2)-containing protein tyrosine phosphatase 2 (Shp2) were developed as a viable method for treating levodopa-induced dyskinesias ( Sellnow et al, 2015 ).…”