NLRP3 gene knockout blocks NF-κB and MAPK signaling pathway in CUMS-induced depression mouse model.

BACKGROUND: Abundant researches indicate that neuroinflammation has important roles in the pathophysiology of depression. Our previous study found that the NLRP3 inflammasome mediated stress-induced depression-like behavior in mice via regulating neuroinflammation. However, it still remains unclear that how the NLRP3 inflammasome influences related inflammatory signaling pathway to contribute to neuroinflammation in depression. METHODS: We used wild-type mice and NLRP3 gene knockout mice to explore the role of NLRP3 inflammasome and related inflammatory signaling pathways in chronic unpredictable mild stress (CUMS) induced depression mouse model. RESULTS: Both wild-type and NLRP3 knockout stress group mice gained less weight than control group mice after 4 weeks CUMS exposure. The wild-type mice subjected to 4 weeks CUMS displayed depression-like behaviors, including decreased sucrose preference and increased immobility time in the tail suspension test. The NLRP3 knockout stress group mice didn't demonstrate depression-like behaviors. The levels of interleukin-1ß protein in serum and hippocampi of CUMS exposed wild-type mice were significantly higher, while the NLRP3 knockout stress group mice didn't show an elevation of interleukin-1ß levels. Similarly to early researches, we found that CUMS led to promoted NLRP3 expression in hippocampi. In addition, the hippocampi in CUMS exposed wild-type mice had higher p-JNK and p-p38 protein expression, which indicated activation of the mitogen-activated protein kinases (MAPK) pathway. The knockout of NLRP3 gene inhibited CUMS-induced activation of the MAPK pathway. The nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) protein complex was activated in the hippocampi of wild-type mice after CUMS exposure, while knockout of NLRP3 gene hindered its activation. CONCLUSIONS: These data further proved that the NLRP3 inflammasome mediated CUMS-induced depression-like behavior. The NLRP3 inflammasome regulated CUMS-induced MAPK pathway and NF-κB protein complex activation in depression mouse model. Further researches targeting the NLRP3 inflammasome-signaling pathway might be under a promising future in the prevention and treatment of depression.

Salvianolic acid B ameliorates depressive-like behaviors in chronic mild stress-treated mice: involvement of the neuroinflammatory pathway.

AIM: Major depressive disorder (MDD) is a debilitating mental disorder associated with dysfunction of the neurotransmitter-neuroendocrine system and neuroinflammatory responses. Salvianolic acid B (SalB) has shown a variety of pharmacological activities, including anti-inflammatory, antioxidant and neuroprotective effects. In this study, we examined whether SalB produced antidepressant-like actions in a chronic mild stress (CMS) mouse model, and explored the mechanisms underlying the antidepressant-like actions of SalB. METHODS: Mice were subjected to a CMS paradigm for 6 weeks. In the last 3 weeks the mice were daily administered SalB (20 mg·kg(-1)·d(-1), ip) or a positive control drug imipramine (20 mg·kg(-1)·d(-1), ip). The depressant-like behaviors were evaluated using the sucrose preference test, the forced swimming test (FST), and the tail suspension test (TST). The gene expression of cytokines in the hippocampus and cortex was analyzed with RT-PCR. Plasma corticosterone (CORT) and cerebral cytokines levels were assayed with an ELISA kit. Neural apoptosis and microglial activation in brain tissues were detected using immunofluorescence staining. RESULTS: Administration of SalB or imipramine reversed the reduced sucrose preference ratio of CMS-treated mice, and significantly decreased their immobility time in the FST and TST. Administration of SalB significantly decreased the expression of pro-inflammatory cytokines IL-1ß and TNF-α, and markedly increased the expression of anti-inflammatory cytokines IL-10 and TGF-ß in the hippocampus and cortex of CMS-treated mice, and normalized their elevated plasma CORT levels, whereas administration of imipramine did not significantly affect the imbalance between pro- and anti-inflammatory cytokines in the hippocampus and cortex of CMS-treated mice. Finally, administration of SalB significantly decreased CMS-induced apoptosis and microglia activation in the hippocampus and cortex, whereas administration of imipramine had no significant effect on CMS-induced apoptosis and microglia activation in the hippocampus and cortex. CONCLUSION: SalB exerts potent antidepressant-like effects in CMS-induced mouse model of depression, which is associated with the inhibiting microglia-related apoptosis in the hippocampus and the cortex.

Antidepressant-like effects of Sanyuansan in the mouse forced swim test, tail suspension test, and chronic mild stress model.

Natural products have been widely reported as effective therapeutic alternatives for treatment of depression. Sanyuansan is a compound recipe composed of ginseng total saponins, fish oil, and valeriana. The aims of this study were to validate whether Sanyuansan has antidepressant-like effects through acute behavioral tests including the forced swimming test (FST), tail suspension test (TST), locomotor activity test, and chronic mild stress (CMS) mice model of depression. C57BL/6 mice were given oral administration of 30 mg/kg imipramine, Sanyuansan, and saline, respectively. The acute behavioral tests including the TST, FST, and locomotor activity test were done after the administration of drugs for consecutively three times (24 hours, 1 hour, and 0.5 hour prior to the tests). Furthermore, the sucrose preference and the serum corticosterone level of mice in the CMS model were examined. Sanyuansan only at 900 mg/kg markedly reduced immobility time in the TST compared with the saline-treated group of mice. Sanyuansan at doses of 225 mg/kg, 450 mg/kg, and 900 mg/kg significantly reduced immobility time of mice in the FST. Sanyuansan reversed the CMS-induced anhedonia and hyperactivation of the hypothalamus-pituitary-adrenal axis. In addition, our results showed that neither imipramine nor Sanyuansan at any dosage increased spontaneous motor activity. These results suggested that Sanyuansan induced significant antidepressant-like effects in mice in both acute and chronic animal models, which seemed unlikely to be attributed to an increase in locomotor activities of mice, and had no sedative-like effects.

[Studies on spray of niclosamide ethanolamine salt. I. development and effect assessment of spray of niclosamide ethanolamine salt].

OBJECTIVE: To develop a spray of niclosamide ethanolamine salt for prevention of bovine from Schistosoma japonicum infection, and explore its characteristics and effect. METHODS: The solubilizers, penetrating agents, emulsifiers were screened, and the spray of niclosamide ethanolamine salt was formulated according to the screening results. The niclosamide ethanolamine salt was determined by using a HPLC technique, and the stability was observed. The preventive effect of the spray was assessed by in-vitro trials against cercariae and protection trials in mice. RESULTS: The screened formulation was presented as following: 1% niclosamide ethanolamine salt was dissolved in 18% dimethyl sulfoxide, and then added with 1% azone and 2% span, together with 78% ethanol, to yield a 1% spray of niclosamide ethanolamine salt. The spray appeared golden flowing liquid, with 1% niclosamide ethanolamine salt in content (W/W), pH 7.4-7.8, and good thermal and cold stability. All cercariae died (100%) while exposed to the spray at a concentration of 1.00 mg/L for 2 min, and the similar effect was achieved while exposed to 0.50 mg/L of the spray for 5 min or 0.10 mg/L for 30 min. The spray at concentrations less than 0.05 mg/L had no evident toxicity to cercariae. A volume of 0.5 ml of the 1% spray was sprayed on the abdomen of mice, 1-3 d later, the mice were infected with S. japonicum cercariae on the spraying sites, no mice were infected, with a protection rate of 100%. Five days post-spraying, the protection rate was 40%, and the worm burden reduction rate was 65.87%. Ten days later, all the mice were infected, however, the worm burden reduction rate was 51.98%. The worm burdens on days 5 or 10 post-spraying were significantly lower than those of the control (P < 0.01). The spray exhibited a good preventive efficacy to mice from S. japonicum infection in lab. CONCLUSIONS: The spray of niclosamide ethanolamine salt has stable physical and chemical property, and is a novel liquid preventive agent against bovine schistosomiasis. In addition, it has a rapid activity against S. japonicum cercariae, so can prevent bovine from S. japonicum infection.

Effects on the expression of GABAA receptor subunits by jujuboside A treatment in rat hippocampal neurons.

AIM OF STUDY: To determine the effect of jujuboside A (JuA) in modulating the gamma-aminobutyric acid (GABA(A)) receptor subunits gene expression of hippocampal neurons at different terms in vitro. MATERIALS AND METHODS: Hippocampal neurons of rat were cultured in vitro, treated with JuA or diazepam (DZP). Then GABA(A) receptor mRNAs were evaluated by semi-quantitative RT-PCR. RESULTS: JuA at the low dose of 41 microM (about 0.05 g/l) induced significant increase of GABA(A) receptor alpha1, alpha5, beta2 subunit mRNAs in both 24 and 72h treatments. JuA at the high dose of 82 microM (about 0.1g/l) significantly increased GABA(A) receptor alpha1, alpha5 subunit mRNA levels and decreased beta2 subunit mRNA level at 24h treatment, and decreased GABA(A) receptor subunit alpha1, beta2 mRNAs expression at 72h treatment. DZP of 10 microM significantly increased expression of GABA(A) receptor subunit alpha1, alpha5 and decreased expression of beta2 at 24h treatment, and decreased alpha1, alpha5, beta2 subunits gene expression at 72h treatment. CONCLUSION: Differences in alterations in GABA(A) receptor subunit mRNAs expression following JuA and DZP treatments could help to explain the differences in the pharmacological action of the two drugs.

Down-regulation of APLP1 mRNA expression in hippocampus of pilocarpine-induced epileptic rats.

OBJECTIVE: To investigate the expression of amyloid beta precursor-like protein 1(APLP1) gene on the transcription level in hippocampus of pilocarpine-induced epileptic rats. METHODS: Epileptic rats were developed by LiC1 (3 mmol/kg, i.p.) approximately 20 h prior to pilocarpine (30 mg/kg, i.p.) administration. The 3' end partial sequence of rat APLP1 gene was cloned, and the expression levels of APLP1 mRNA in hippocampus of epileptic rats at 6 h, 30 h, 7 d and 15 d were determined by semi-quantitative RT-PCR. RESULTS: The 3'end partial sequence of rat APLP1 gene shared a 97% homology with that of mice, and 90% with that of human. The APLP1 amino acid sequence of rat was identical with that of mouse, but was different from that of human in 3 residues. Moreover, pilocarpine induced a significant down-regulation of APLP1 mRNA expression at 6 h after epilepsy initiation (P< 0.05), and at 30 h, this down-regulation became more dramatic (P< 0.01), which lasted till day 15 (P< 0.01). CONCLUSION: The 3' end of APLP1 gene is highly conserved, and APLP1 mRNA expression is kept at low level in hippocampus of pilocarpine-induced epileptic rats.

Upregulation of Mark3 and Rpgrip1 mRNA expression by jujuboside A in mouse hippocampus.

AIM: To investigate the effect of jujuboside A (JuA) on modulating gene expression in the hippocampus. METHODS: The spontaneous activity of mice was monitored, and the differential display polymerase chain reaction was adapted to screen differentially-expressed genes modulated by JuA in the mouse hippocampus. RESULTS: JuA significantly decreased the total activity intensity (P<0.01 vs control) at a dosage of 80 mg/kg, and the genes MAP/microtubule affinity-regulating kinase3 (Mark3) and retinitis pigmentosa GTPase regulator interacting protein1 (Rpgrip1) were upregulated by JuA in the mouse hippocampus. CONCLUSION: JuA had an inhibitory effect on the spontaneous activity of the mice, and JuA regulated the transcription of Mark3 and Rpgrip1 in the mouse hippocampus.

3,4-oxo-isopropylidene-shikimic acid inhibits adhesion of polymorphonuclear leukocyte to TNF-alpha-induced endothelial cells in vitro.

AIM: To examine the effect of 3,4-oxo-isopropylidene-shikimic acid (ISA) on human polymorphonuclear leukocyte (PMN) adhesion to human umbilical vein endothelial cells (HUVEC) and explore its mechanism. METHODS: Adhesion of PMN to HUVEC was measured by rose bengal staining assay. Cell-ELISA and RT-PCR methods were used to examine the expression of adhesion molecules ICAM-1. Cell viability was detected with MTT assay. RESULTS: ISA (1-100 micromol/L) effectively reduced PMN adhesion to TNF-alpha-induced HUVEC with the inhibitory rate from 17.2 % to 53.5 %, and exerted no effect on PMN adhesion to normal HUVEC. Adhesion molecule ICAM-1 surface protein and mRNA expression induced by TNF-alpha (400 kU/L) were significantly inhibited by ISA. In addition, the cell viability of HUVEC was unchanged 48 h after treatment with ISA. CONCLUSION: ISA inhibited TNF-alpha-stimulated PMN-HUVEC adhesion and expression of ICAM-1.