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Abstract Background: Phospholipase C-like 1 (PLCL1), a protein that lacks catalytic activity, has similar structures to the PLC family. The aim of this research was to find the function and underlying mechanisms of PLCL1 in fibroblast-like synoviocyte (FLS) of rheumatoid arthritis (RA). Methods: In this study, we first analyzed the expression of PLCL1 in the synovial tissue of RA patients and K/BxN mice by immunohistochemical staining. Then silencing or overexpressing PLCL1 in FLS before stimulating by TNF-α. The levels of IL-6, IL-1β and CXCL8 in FLS and supernatants were detected by Western Blot (WB), Real-Time Quantitative PCR and Enzyme Linked Immunosorbent Assay. We used INF39 to specifically inhibit the activation of NLRP3 inflammasomes, and detected the expression of NLRP3, Cleaved Caspase-1, IL-6 and IL-1β in FLS by WB. Result: When PLCL1 was silenced, the level of IL-6, IL-1β and CXCL8 were down-regulated. When PLCL1 was overexpressed, the level of IL-6, IL-1β and CXCL8 were unregulated. The previous results demonstrated that the mechanism of PLCL1 regulating inflammation in FLS was related to NLRP3 inflammasomes. INF39 could counteract the release of inflammatory cytokines caused by overexpression of PLCL1. Conclusion: Result showed that the function of PLCL1 in RA FLS might be related to the NLRP3 inflammasomes. We finally confirmed our hypothesis with the NLRP3 inhibitor INF39. Our results suggested that PLCL1 might promote the inflammatory response of RA FLS by regulating the NLRP3 inflammasomes.
Résumé
Objective:To investigate the neuroprotective effect of Danggui Shaoyaosan （DSS） in a rat model of amyloid-<italic>β</italic>-peptide<sub>1-42</sub> （A<italic>β</italic><sub>1-42</sub>）-induced Alzheimer's disease （AD） as well as its regulatory effect on NOD-like receptor protein 3 （NLRP3）/cysteinyl aspartate-specific protease-1 （Caspase-1） signaling pathway. Method:The AD animal model was established via intracerebral injection of A<italic>β</italic><sub>1-42</sub> and treated with different concentrations of DSS after the division of rats into the sham operation group， model group， as well as the high-， medium-， and low-dose DSS groups. Morris water maze test was conducted to determine the learning and memory abilities of rats. The morphology and function of neurons were detected by hematoxylin-eosin （HE） staining and Golgi staining， followed by immunofluorescence co-localization of NLRP3 inflammasome activation. The mRNA expression levels of interleukin （IL）-1<italic>β</italic> and IL-18 were measured by Real-time polymerase chain reaction （Real-time PCR）， and the protein expression levels of NLRP3， Caspase-1， and IL-1<italic>β </italic>were assayed by Western blot. Result:Compared with the sham operation group， the model group exhibited significantly decreased learning and memory abilities （<italic>P</italic><0.01）， impaired neuronal morphology and function， up-regulated IL-1<italic>β</italic> and IL-18 mRNA expression， enhanced NLRP3 inflammasome activation， and elevated NLRP3， Caspase-1， and IL-1<italic>β</italic> protein expression （<italic>P</italic><0.01）. Compared with the model group， DSS at both medium and high doses remarkably improved the learning and memory abilities of AD rats （<italic>P</italic><0.05， <italic>P</italic><0.01）， restored neuronal morphology and function， down-regulated the mRNA expression levels of inflammatory factors IL-1<italic>β</italic> and IL-18， reduced the activation of NLRP3 inflammasomes， and lowered the protein expression levels of NLRP3， Caspase-1， and IL-1<italic>β</italic> （<italic>P</italic><0.01）. Conclusion:DSS inhibits inflammasome activation and neuroinflammatory response possibly by regulating the NLRP3/Caspase-1 signaling pathway， thus exerting the neuroprotective effect.
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Objective To investigate the mechanisms of paeoniflorin in protection of retinal ischemia injury.Methods Fifty-four male specefic pathogen free (SPF) degree Wistar rats were randomly divided into normal control group,model control group and paeoniflorin group.Retinal ischemia injury was induced by raising the intraocular pressure of right eyes of rats to 110 mmHg for 30 minutes.The rats of paeoniflorin group were administrated through intraperitoneal injection of 5 mg/kg paeoniflorin each day for 14 days.OCT and electroretinogram (ERG) were performed to detect the thickness of retinal nerve fiber layer+retinal ganglion cell layer+inner plexiform layer (NGI)and electrophysiological changes of retina,respectively.Retrograde labelling of retinal ganglion cells (RGCs) was used to evaluate the survival number of RGCs.Western blot analysis was used to detect NLRP3,apoptosis-associated speck-like protein containing a caspase activation and recruitment domain (ASC),cleaved caspase 1 (c-caspase 1),IL-18,and IL-1β expression.The use and care of animals complied with the statement of the Association for Research in Vision and Ophthalmology (ARVO) and Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results The thickness of retinal NGI in model control group was (58.2 ± 1.7) μm,which was significantly lower than (84.8 ± 1.9) μm in normal control group and (71.1 ±2.4) μm in paeoniflorin group (both at P＜0.05).The amplitudes of A and B waves in paeoniflorin group and normal control group were significantly higher than those in model control group (both at P＜0.05).The number of RGC in model control group was significantly lower than that in paeoniflorin group and normal control group (both at P＜0.05).The relative expressions of NLRP3,ASC,c-caspase 1,IL-18 and IL-1β in model control group were significantly higher than those in normal control group and paeoniflorin group (all at P＜0.05).Conclusions The paeoniflorin can prevent retinal ischemia induced injury of the retina through NLRP3 inflammasomes pathway,which provides a new treatment strategy for clinical therapy.
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The aim of this study was to explore the neuroprotective effect and mechanism of XingNaoJing injections (XNJ) on cerebral ischemia injury and blood-brain barrier (BBB) disruption. Middle cerebral artery occlusion (MCAO) method was applicated to establish the model of cerebral ischemia/reperfusion (I/R) injury in rats. BBB permeability after I/R injury was assessed with the leaking amount of Evans Blue and the expression of occludin and ZO-1. The expression of NOD-like receptor family, pyrin domain containing (NLRP3) was checked to explore the inhibition of inflammation by XNJ. The results showed that XNJ could significantly increase the survival percent, decrease the infarct area and ameliorate neurological deficits and brain damage after I/R injury. Leaking amount of Evans Blue was reduced by XNJ, and the expression of tight junction protein, occludin and ZO-1 was also up-regulated by XNJ, which showed a role of protection on BBB disruption. The expression of NLRP3 was inhibited after exposure of XNJ, which was associated with inhibition of the inflammatory response. In summary, XNJ could suppress NLRP3 inflammasomes and improve BBB disruption and brain damage in rats after cerebral I/R injury, which provided a beneficial insight to further explore XNJ.
Résumé
Objective To investigate the mechanisms of paeoniflorin in protection of retinal ischemia injury. Methods Fifty.four male specefic pathogen free ( SPF) degree Wistar rats were randomly divided into normal control group,model control group and paeoniflorin group. Retinal ischemia injury was induced by raising the intraocular pressure of right eyes of rats to 110 mmHg for 30 minutes. The rats of paeoniflorin group were administrated through intraperitoneal injection of 5 mg/kg paeoniflorin each day for 14 days. OCT and electroretinogram ( ERG ) were performed to detect the thickness of retinal nerve fiber layer+retinal ganglion cell layer+inner plexiform layer ( NGI) and electrophysiological changes of retina, respectively. Retrograde labelling of retinal ganglion cells ( RGCs ) was used to evaluate the survival number of RGCs. Western blot analysis was used to detect NLRP3,apoptosis.associated speck.like protein containing a caspase activation and recruitment domain (ASC),cleaved caspase 1 (c.caspase 1), IL.18,and IL.1β expression. The use and care of animals complied with the statement of the Association for Research in Vision and Ophthalmology ( ARVO ) and Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission. Results The thickness of retinal NGI in model control group was ( 58. 2 ± 1. 7)μm, which was significantly lower than ( 84. 8 ± 1. 9)μm in normal control group and(71. 1±2. 4)μm in paeoniflorin group (both at P<0. 05). The amplitudes of A and B waves in paeoniflorin group and normal control group were significantly higher than those in model control group ( both at P<0. 05 ) . The number of RGC in model control group was significantly lower than that in paeoniflorin group and normal control group ( both at P<0. 05). The relative expressions of NLRP3,ASC,c.caspase 1,IL.18 and IL.1β in model control group were significantly higher than those in normal control group and paeoniflorin group (all at P<0. 05). Conclusions The paeoniflorin can prevent retinal ischemia induced injury of the retina through NLRP3 inflammasomes pathway,which provides a new treatment strategy for clinical therapy.