Haijie Ji

Osthole improves chronic cerebral hypoperfusion induced cognitive deficits and neuronal damage in hippocampus.

This study is to investigate the effects of osthole on cognitive impairment and neuronal degeneration in hippocampus induced by chronic cerebral hypoperfusion in rats, as well as the potential mechanism. Permanent occlusion of bilateral common carotid arteries (2VO) induced severe cognitive deficits tested by the water maze task, along with oxidative stress and neuronal loss in hippocampus. Oral administration of osthole for 3 weeks markedly attenuated cognitive deficits and neuronal damage. Biochemical experiments revealed that osthole decreased the production of malondialdehyde (MDA) and significantly increased the activities of Glutathione Peroxidase (GPx) and Catalase. Western blot analyses indicated that osthole prevented the downregulation of bcl-2 expression and upregulation of bax expression, which resulted in decreasing bax/bcl-2 ratio in hippocampus of 2VO rats. Additionally, osthole effectively alleviated the activation of caspase-3 induced by permanent occlusion of bilateral common carotid arteries. The observed results in present study suggest that osthole exhibits therapeutic potential for vascular dementia, which is most likely related, at least in part, to its antioxidation and anti-apoptotic actions.

Discovery and Optimization of Novel 3-Piperazinylcoumarin Antagonist of Chemokine-like Factor 1 with Oral Antiasthma Activity in Mice

Chemokine-like factor 1 (CKLF1) is a novel functional cytokine that acts through its receptor CC chemokine receptor 4 (CCR4). Activation of CCR4 by CKLF1 plays an important role in diseases such as asthma and multiple sclerosis. This article describes a cell-based screening assay using an FITC-labeled CCR4 agonist (CKLF1-C27), a CKLF1 peptide fragment. Screening of our in-stock small-molecule library identified a 3-piperazinylcoumarin analogue 1 (IC(50) = 4.36 x 10(-6) M) that led to the discovery of orally active compound 41 (IC(50) = 2.12 x 10(-8) M) through systematic optimization. Compound 41 blocked the calcium mobilization and chemotaxis induced by CKLF1-C27 and reduced the asthmatic pathologic changes in lung tissue of human CKLF1-transfected mice. Further studies indicated that compound 41 ameliorated pathological changes via inhibition of the NF-kappaB signal pathway.

Inhibitory effect of ginsenoside Rg1 on lipopolysaccharide-induced microglial activation in mice.

Microglial activation plays a pivotal role in the pathogenesis of neurodegenerative diseases by producing various pro-inflammatory cytokines and nitric oxide (NO). In the paper, the anti-inflammatory effect of ginsenoside Rg1 was investigated in mice intracerebroventricular injected of lipopolysaccharide (LPS). NO and tumor necrosis factor (TNF)-α production in both cerebral cortex and hippocampus decreased at dose-dependent manner by oral administration with Rg1. And the expression of ionized calcium binding adaptor molecule 1 (Iba-1) increased in both cerebral cortex and hippocampus in LPS-injected group compared to that in control group. However, Rg1 inhibited microglial activation by suppressing Iba-1 expression. In addition, the expression of inducible nitric oxide synthase (iNOS) was inhibited by Rg1. Moreover, Rg1 suppressed the phosphorylation level of IκB, nuclear translocation of p65 subunit of NFκB, and phosphorylation level of p38, ERK1/2, JNK mitogen-activated protein kinase (MAPK) induced by LPS. Concluding, Rg1 inhibited the inflammation mediated by LPS by suppressing NFκB and MAPK pathway, which provided the explanation for its therapeutic effect on neurodegenerative diseases.

Effect of compound IMMLG5521, a novel coumarin derivative, on carrageenan-induced pleurisy in rats.

Accumulative evidences have showed that some coumarin derivatives have significantly anti-inflammatory effects. To investigate the potential anti-inflammatory effect of compound IMMLG5521, a novel coumarin derivative, carrageenan-induced pleurisy model in rats was employed. The results showed that IMMLG5521 (5, 10 and 20 mg/kg) exhibited anti-inflammatory effects, reducing pleural exudate formation, decreasing total number of inflammation cells and polymorphonuclear leukocytes infiltration, attenuating histological injury and reducing TNF-α, IL-1β, MIP-2 and IL-8 release. Further investigation revealed that the compound may exert its anti-inflammatory effect via inhibiting nuclear translocation of NF-кB in inflammatory cells collected from pleural exudates. Taken together, the present results suggested that IMMLG5521 inhibited acute inflammation in carrageenan-induced pleurisy model that could be, in part, related to a reduction of release of inflammatory factors, another part may be related to an inhibition of NF-кB activation.

Anti-inflammatory effect of IMMLG5521, a coumarin derivative, on Sephadex-induced lung inflammation in rats

The study is to investigate the effect of compound IMMLG5521, a coumarin derivative, on lung inflammation induced by Sephadex in rats. Sephadex led to massive granulomas, infiltration of neutrophils and eosinophils, the increase of TNF-α level in BALF and lung tissue. Sephadex injection led to the up-regulation of VCAM-1 and ICAM-1 expressions in the lungs. However, pretreatment with IMMLG5521 (6.25 and 12.5 mg/kg) inhibited massive granulomas and infiltration of neutrophils and eosinophils, decreased TNF-α level in BALF and lung tissue, and down-regulated VCAM-1 and ICAM-1 expressions in the lung tissue. In conclusion, compound IMMLG5521 may suppress the lung injury induced by Sephadex, at least in part, due to the prevention of the up-regulation of VCAM-1 and ICAM-1 expressions and TNF-α level.

IMM-H004, a novel courmarin derivative, protects against oxygen-and glucose-deprivation/restoration-induced apoptosis in PC12 cells

7-Hydroxy-5-methoxy-4-methyl-3-(4-methylpiperazin-1-yl)-coumarin (IMM-H004) is a novel coumarin derivative synthesized in our laboratory. The purpose of the current study was to determine the neuroprotective effects of IMM-H004 on PC12 cells and its potential mechanism of action. PC12 cells were subject to oxygen and glucose deprivation (OGD) followed by the restoration of oxygen and glucose (R), which mimics ischemia and reperfusion in vivo. Cell viability was determined by MTT assay. DNA fragmentation was analyzed by DNA ladder. ROS and mitochondrial membrane potential were measured by fluorescent microscope and quantified by Image-Pro Express 6.0 software. ATP was measured by luciferin-luciferase assay. The activation of signal-regulated molecules was assessed by the Western blot analysis. OH formation was determined using the Electron Spin Resonance (ESR) trapping technique in combination with 5, 5-dimethyl-1-pyrroline-N-oxide. OGD/R reduced cell viability and induced cell apoptosis, which were both dose-dependently attenuated by IMM-H004. The accumulation of intracellular reactive oxygen species (ROS) and reduced mitochondrial membrane potential observed in PC12 cells treated with OGD/R, which switch on the mitochondrion-dependent apoptotic pathway, were reversed by IMM-H004. ATP production in OGD/R-treated PC12 cells was elevated by IMM-H004, which suggests that it restored the functions of the mitochondria. OGD/R-induced cytochrome c release from the mitochondria reduced the ratio of apoptotic proteins, Bcl-2/Bax, and induced caspase-3 activation and DNA fragmentation. These changes were significantly inhibited by IMM-H004. IMM-H004 also significantly inhibited OH formation, determined by electron spin resonance, which indicates that it is a potent free-radical scavenger. This study has demonstrated that IMM-H004 protects PC12 cells against OGD/R-induced apoptosis, at least in part, by scavenging excessive ROS and inhibiting the mitochondrion-dependent apoptotic pathway.

Osthole attenuates the development of carrageenan-induced lung inflammation in rats

Osthole has been reported to possess a variety of pharmacological activities, such as antiinflammatory effect. In the present study, we have investigated the effect of osthole on lung inflammation associated with carrageenan-induced pleurisy in rats. The result showed that osthole could inhibit significantly pleural exudates formation and PMNs infiltration. Histological examination revealed osthole could reduce lung inflammation in rats treated with carrageenan. The myeloperoxidase (MPO) level was examined in pleural exudates. The result showed that osthole could attenuate MPO level in pleural exudates. Further studies showed osthole could decrease tumor necrosis factor alpha (TNF-α) and interleukin 1beta (IL-1β) levels in the lungs. Taken together, the present results suggested that osthole could inhibit lung inflammation on carrageenan-induced pleurisy in rats and that could be related to a reduction of PMNs infiltration and release of inflammatory factors.

In vitro and in vivo anti-inflammatory effects of IMMLG5521, a coumarin derivative.

Several coumarin derivatives have been reported to present multiple biological activities. In this study, in vitro the compound IMMLG5521 (0.1, 1, 10μM) can inhibit the release of β-glucuronidase from PAF-stimulated polymorphonuclear leukocytes, the compound IMMLG5521 (0.1, 1, 10μM) can inhibit NO production and decrease TNF-α and IL-β release from LPS-stimulated RAW264.7 cells. In vivo, we evaluated the effect of IMMLG5521 on acute and chronic inflammation models. Our data showed that IMMLG5521 (6mg/kg, 12mg/kg) could inhibit xylene-induced ear swelling and cotton pellet-induced granuloma formation in mice. Taken together, the compound IMMLG5521 inhibited the release of inflammatory factors and mediators in vitro, decreased inflammation response in mice. The compound IMMLG5521 can inhibit inflammation in vitro and in vivo.

The chemokine-like factor 1 induces asthmatic pathological change by activating nuclear factor-κB signaling pathway.

CKLF1, which exhibits chemotactic activities on a wide spectrum of leukocytes, is up-regulated during the progress of asthma. It plays a vital role in the pathogenesis of pulmonary disease. Here, we report that CKLF1 has the capability to activate the NF-κB signaling pathway leading to the pathological change in the lung. The HEK293-CCR4 cell line, which expressed CCR4 stably, was established and screened. Western blot analysis was performed to determine the expression of NF-κB in HEK293-CCR4 and A549 cells following the C27 (10μg/ml) added in each well at different times. These results showed that C27 (10μg/ml) time-dependently induced the accumulation of NF-κB in the nucleus of HEK293-CCR4 and A549 cells. In addition, CKLF1 plasmid (100μg) injection and electroporation led to the asthmatic change in the lung in mice as shown by HE and PAS staining. Furthermore, it was confirmed that CKLF1 significantly up-regulated the p-IκB expression, decreased the IκB expression, and suppressed the NF-κB expression in the cytoplasm of pulmonary tissue in vivo study. Intriguingly, an enhanced nuclear accumulation of NF-κB was observed in the lung of pCDI-CKLF1 electroporated mice, compared to that in the sham group. Therefore, the NF-κB signaling pathway was involved in the asthmatic change induced by CKLF1, among which CCR4 might play a crucial role.

Study on the Dynamic Changes in Synaptic Vesicle-Associated Protein and Axonal Transport Protein Combined with LPS Neuroinflammation Model

Microglia activation is the major component of inflammation that constitutes the characteristic of neurodegenerative disease. A large amount of researches have demonstrated that inflammation involved in the pathogenesis of PD process activated microglia acting on the neurons through the release of a variety of inflammatory factors. However, the molecular mechanism underlying how it does work on neurons is still unclear. Here, we show that intracerebral injections of LPS induced Parkinsons disease pathology in C57BL/6J mice. Furthermore, study on the dynamic changes in Synaptic vesicle-associated protein and axonal transport Protein in this process. The results indicated that after administration of LPS in the brain, the inflammatory levels of TNF-α and IL-1β both are elevated, and have a time-dependent.