Therapeutic Effect of Ziziphi Spinosae Semen-Albiziae Flos on Depression in Rats / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Ziziphi Spinosae Semen-Albiziae Flos on the nucleotide-binding oligomerization domain，NOD-like receptor thermal protein domain associated protein 1 （NLRP1）/chemokine ligand 1 （CXCL1）/chemokine receptor 2 （CXCR2） pathway in the hippocampus of the rat model of chronic unpredictable mild stress （CUMS）-induced depression. MethodA total of 120 male SD rats were randomized into blank，CUMS，CUMS + low-，medium-，and high-dose （4，8，16 g·kg-1） Ziziphi Spinosae Semen-Albiziae Flos，and CUMS + venlafaxine hydrochloride （0.008 g·kg-1） groups，with 20 rats in each group.The rat model of depression was established by solitary feeding combined with CUMS.The behaviors and spatial learning and memory abilities of rats were examined by sugar water consumption test，tail suspension test，forced swimming test，and Morris water maze test.Quantitative real-time PCR （Real-time PCR） and Western blot were employed to determine the expression of factors associated with the NLRP1/CXCL1/CXCR2 pathway in the hippocampus.Enzyme-linked immunosorbent assay was employed to determine the levels of tumor necrosis factor-α （TNF-α），interleukin （IL）-18，IL-1β，and IL-6 in the hippocampus.The immunofluorescence assay was used to measure the levels of reactive oxygen species （ROS） in the hippocampus. ResultCompared with the blank group，the CUMS group showed decreased preference to sugar water and times of crossing the platform （P<0.01），and increased immobility time of tail suspension，forced swimming floating time，and escape latency （P<0.01）.Compared with the CUMS group，the administration of Ziziphi Spinosae Semen-Albiziae Flos and venlafaxine hydrochloride alleviated the effects of CUMS on the above-mentioned behaviors and spatial learning and memory abilities of the rats （P<0.05，P<0.01）.Compared with the blank group，the CUMS group showed up-regulated protein levels of NLRP1，CXCL1，and CXCR2 （P<0.01） and elevated levels of IL-18，IL-1β，TNF-α，and IL-6 （P<0.01） in the hippocampus.The treatment with Ziziphi Spinosae Semen-Albiziae Flos and venlafaxine hydrochloride attenuated the activation of NLRP1/CXCL1/CXCR2 signaling pathway and lowered the levels of inflammatory cytokines in the hippocampus of CUMS rats （P<0.05，P<0.01）.In addition，Ziziphi Spinosae Semen-Albiziae Flos lowered the level of ROS in the hippocampus （P<0.05，P<0.01）. ConclusionZiziphi Spinosae Semen-Albiziae Flos can mitigate the depressive behaviors of the rat model of CUMS-induced depression by inhibiting the activation of NLRP1/CXCL1/CXCR2 signaling pathway.

Azeliragon ameliorates Alzheimer's disease via the Janus tyrosine kinase and signal transducer and activator of transcription signaling pathway

OBJECTIVES: TTP488, an antagonist of the receptor for advanced glycation end-products, was evaluated as a potential treatment for patients with mild-to-moderate Alzheimer's disease (AD). However, the mechanism underlying the protective action of TTP488 against AD has not yet been fully explored. METHODS: Healthy male rats were exposed to aberrant amyloid β (Aβ) 1-42. Lipopolysaccharide (LPS) and the NOD-like receptor family pyrin domain containing 1 (NLRP1) overexpression lentivirus were injected to activate the NLRP1 inflammasome and exacerbate AD. TTP488 was administered to reverse AD injury. Finally, tofacitinib and fludarabine were used to inhibit the activity of Janus tyrosine kinase (JAK) and signal transducer and activator of transcription (STAT) to prove the relationship between the JAK/STAT signaling pathway and TTP488. RESULTS: LPS and NLRP1 overexpression significantly increased the NLRP1 levels, reduced neurological function, and aggravated neuronal damage, as demonstrated by the impact latency time of, time spent by, and length of the platform covered by, the mice in the Morris water maze assay, Nissl staining, and immunofluorescence staining in rats with AD. CONCLUSIONS: TTP488 administration successfully reduced AD injury and reversed the aforementioned processes. Additionally, tofacitinib and fludarabine administration could further reverse AD injury after the TTP488 intervention. These results suggest a new potential mechanism underlying the TTP488-mediated alleviation of AD injury.

Ginsenoside Rgl regulates NOX2-NLRP1 pathway to reduce hypoxia-reoxygenation injury in PC12 cells / 中国药理学通报

Aim To investigate the effect of ginsenoside Rgl on PC 12 cell hypoxia-reoxygenation injury and its possible mechanism. Methods PC 12 cells were randomly divided into six groups. Except for the blank control group, all the other groups were hypoxia and hypoglycemia for 6 hours, and then reoxygenated and glycosylated for 24 hours to make OGD/R models. Each drug group was given corresponding drugs 2 hours before modeling pretreatment. DCFH-DA method was used to detect the ROS production in cells, Annexin V- FITC/PI double staining method was performed to detect cell apoptosis rate, ELISA method was used to detect LDH activity and IL-1 (3 content in cell supernatant, and Western blot was applied to detect the ex- pression of proteins of N0X2, p22phox, p47phox, NLRPl, ASC, Caspase-1, PSD95, Tau, p-Tau and observe the intervention effect of ginsenoside Rgl. Re sults Tempol, Apocynin and Rgl (5, 10 jjLmol • L"1) groups could significantly inhibit ROS production and apoptosis, reduce LDH release and IL-1 (3 content in cell supernatant; Apocynin and Rgl (5, 10 |xmol • L"1) groups could significantly down-regulate the expression of N0X2, p22phox and p47phox in cells. The Tempol, Apocynin and Rgl (5, 10 jxmol • L"1 ) groups could significantly decrease the protein expression of NLRP1, Caspase-1, ASC, IL-1 (3 and p-Tau, and markedly down-regulate the expression of PSD95 protein. Conclusion Rgl is likely to reduce the is- chemia-reperfusion injury of PC 12 cells by inhibiting the NOX2-NLRP1 pathway.

Expression and clinical significance of NLRP1 and NLRP3 in colonic tissues of patients with ulcerative colitis / 西安交通大学学报(医学版)

【Objective】 To investigate the expressions of NLRP1 and NLRP3 in the colon of ulcerative colitis (UC) patients and analyze the correlation of the expressions with severity of UC, endoscopic manifestations and related laboratory indicators. 【Methods】 We collected biopsical specimens obtained with colonoscopy in 46 patients with UC (22 mild cases and 24 moderate to severe cases) and 20 cases of normal control group. We used the disease activity index to evaluate the Mayo UC inflammatory activity and immunohistochemical method to detect the protein expression levels of intestinal mucosal NLRP1 and NLRP3 in the tissue. RT-PCR was used to detect the expressions of NLRP1 and NLRP3 mRNA in intestinal mucosal tissues. Meanwhile, the colonoscopy, serum uric acid, C-reactive protein, serum sedimentation rate, platelet count, low-density lipoprotein, and cholesterol of UC patients were also counted to further analyze the relationship between NLRP1 and NLRP3. 【Results】 The expressions of NLRP1 and NLRP3 protein and mRNA in colonic mucosal tissues of UC patients were significantly higher than those of normal controls (P<0.05). Compared with that in mild UC, the expression of NLRP1 in colonic mucosal tissues of moderate and severe UC patients was significantly increased (P<0.05). There was no significant difference in the expressions of NLRP1 and NLRP3 in colonic mucosal tissues of UC patients with different lesion ranges. NLRP1 expression was positively correlated with Mayo overall score, Mayo endoscopic score, erythrocyte sedimentation rate, and C-reactive protein (P<0.05), NLRP3 expression was positively correlated with C-reactive protein (P<0.05), but not correlated with Mayo overall score, Mayo endoscopic score, or erythrocyte sedimentation rate. NLRP1 expression was positively correlated with low-density lipoprotein and platelet (P<0.05), but not with uric acid or cholesterol. NLRP3 was positively correlated with low-density lipoprotein, uric acid and cholesterol (P<0.05), but not with platelet. 【Conclusion】 NLRP1 and NLRP3 may be involved in the pathogenesis of UC and related to disease activity. Therefore, they can be used as molecular targets for targeted therapy, and NLRP1 can be used as a predictor of mucosal healing.

Protective effect and mechanism of ginsenoside Rg1 on H2O2induced hippocampal neurons aging due to down-regulate NOX2 mediated NLRP1 inflammasome activation in vitro / 中国药理学与毒理学杂志

OBJECTIVE To explore the protective effects and mechanisms of Ginsenoside Rg1 (Rg1) on H2O2-induced hippocampal neurons aging in vitro. METHODS The primary culture hippo-campal neurons(7 d)were randomly placed into six groups:normal control group,H2O2(200 μM)treat-ment group,and H2O2+Rg1(1,5 and 10μM)groups.The neurons were with Rg1(1,5 and 10 μmol·L-1) for 6h. H2O2(200 μmol·L-1) was added to the medium and incubate for 18 h. The Dihydroethidium (DHE) staining was performed for ROS production assessment. The LDH release and Hoechst 33258 were performed to examine the neuronal damage and apoptosis. The immunoblot was used to deter-mine the expression of β-Gal,NOX2,p22phox,p47phox,NLRP-1,ASC and Caspase-1 in hippocampal neurons.The ELISA was performed to detect the levels of IL-1β and IL-18 released in the supernatant in hippocampal neurons.RESULTS Rg1(5 and 10 μmol·L-1)significantly reduced the ROS production, attenuated H2O2-induced neuronal damage and apoptosis (P<0.05, P<0.01). The immunoblot results showed that Rg1(5 and 10 μmol·L-1)treatment significantly decreased the expression of β-Gal,NOX2, p22phox,p47phox,NLRP-1,ASC and Caspase-1 in hippocampal neurons(P<0.05,P<0.01).Additionally, Rg1(5 and 10 μmol·L-1)treatment significantly decreased IL-1β and IL-18 release in the supernatant. CONCLUSION The protective effect of Rg1 in H2O2-induced hippocampal neurons aging may be due to inhibit NOX2-NLRP1 activation.

Research progresses and the effection of traditional Chinese medicine of pyroptosis and Alzheimer disease / 医学研究生学报

Pyroptosis is a new type of programmed cell death .Pyroptosis tightly controls inflammatory response and destroys cell membranes, which finally induces cell death. From the current knowledge of the role of pyroptosis in Alzheimer disease, we can see that there are several connections between pyroptosis and Alzheimer disease. Frist of all, Inflammasome might play a major role in triggering Amyloid plaques-clumps of protein accumulation. Second, pyroptosis might activate caspase-1 for the subsequent maturation of pro-inflammatory cytokines,IL-1β and IL-18, which could be implicated in the execution of inflammatory response. Third, pyroptosis leads to neuronal cell death which is associated with pathogenetic mechanisms in Alzheimer disease. The goal of this article is to discuss the connection between Pyroptosis and Alzheimer disease so to find out the new therapeutic strategies targeting Alzheimer Disease.

Effects of saponins from Panax japonicus on neuronal apoptosis of natural aging rats by NLRP1 and NLRP3 inflammasome pathway / 中草药

Objective To observe the effects of saponins from Panax japonicus on neuronal apoptosis of natural aging rats and its mechanisms based on NLRP1 and NLRP3 inflammasome pathway. Methods Male SD rats in a SPF grade were randomly divided into five groups: control group (9-month-old rats), model group (24-month-old rats), and SPJ treatment group (10, 30, and 60 mg/kg). From the beginning of 18 months, animals were treated with SPJ (or normal saline) by ig until 24 months, and stopped 2 d each week for six months of continuous administration. The neural apoptosis situation of cortex and hippocampus in aging rats were observed by TUNEL method. The protein expression of IL-1β, ASC, NLRP3, NLRP1, Caspase-1, and IL-18 of the cerebral cortex and hippocampal were detected by Western blotting. Results TUNEL results showed that there were a very small number of apoptotic cells in the cortex and hippocampus in control group. Compared with control group, the model group significantly increased the number of apoptotic cells. Compared with model group, the number of apoptotic cells was significantly decreased in rat cortex and hippocampus (CA1, CA3, and DG) after treated with SPJ (10, 30, and 60 mg/kg). Western blotting results showed a significant age-related increase in the expression of IL-1β, ASC, NLRP3, NLRP1, Caspase-1, and IL-18, while SPJ concentration-dependently decreased the levels of IL-1β, ASC, NLRP3, NLRP1, Caspase-1, and IL-18 after six-month treatment. Conclusion In conclusion, saponins from P. japonicus has protective effects on the brain (cortex and hippocampus) of aging rats. The mechanism is likely to be that saponins from P. japonicus can reduce nerve inflammation by regulating NLRP1 and NLRP3 inflammasome pathway.

Role of SREBP1 in atorvastatin-induced reduction of NLRP1 inflammasome ex-pression / 中国免疫学杂志

Objective:To investigate the role of sterol regulatory element binding protein-1 (SREBP1) in atorvastatin-induced reduction of nucleotide-binding oligomerization domain-like receptor protein 1 ( NLRP1 ) inflammasome expression. Methods:THP-1 cells were treated with phorbol 12-myristate 13-acetate (160 nmol/L) for 12 h to be differentiated into macrophages. The medium was then replaced with serum-free medium containing lipopolysaccharide and ( or ) atorvastatin. The mRNA expression of NLRP1 and SREBP1 were detected by Real-time PCR. The protein expression of NLRP1 and SREBP1 were determined by Western blot. Furthermore, we observed the effect of SREBP1 siRNA on atorvastatin-induced reduction of NLRP1 expression. Results:Atorvastatin inhibited the mRNA and protein expression of NLRP1 and SREBP1 in the THP-1 macrophages. SREBP1 siRNA showed no significant difference on lowering NLRP1 expression when compared with atorvastatin. Treating cells with SREBP1 siRNA and atorvastatin at the same time resulted in more obvious reduction of NLRP1 expression than single use of SREBP1 siRNA or atorvastatin. Conclusion:Atorvastatin might exert anti-inflammatory effect by repressing NLRP1 expression through the SREBP1 path-way.

NLRP3 polymorphism is associated with protection against human T-lymphotropic virus 1 infection

Inter-individual heterogeneity in the response to human T-lymphotropic virus 1 (HTLV-1) infection has been partially attributed to host genetic background. The antiviral activity of the inflammasome cytoplasmic complex recognises viral molecular patterns and regulates immune responses via the activation of interleukin (IL)-1 family (IL-1, IL-18 and IL-33) members. The association between polymorphisms in the inflammasome receptors NLRP1 and NLRP3 and HTLV-1 infection was evaluated in a northeastern Brazilian population (84 HTLV-1 carriers and 155 healthy controls). NLRP3 rs10754558 G/G was associated with protection against HTLV-1 infection (p = 0.012; odds ratio = 0.37). rs10754558 affects NLRP3 mRNA stability; therefore, our results suggest that higher NLRP3 expression may augment first-line defences, leading to the effective protection against HTLV-1 infection.

Role of NLRP3 and NLRP1 inflammasomes signaling pathways in pathogenesis of rheumatoid arthritis

OBJECTIVES@#To investigate the role of NLRP3 and NLRP1 inflammasomes signaling pathways in rheumatoid arthritis (RA).@*METHODS@#A total of 36 patients with RA were selected, peripheral blood mononuclear cell (PBMC) and granulocyte were separated from venous blood. RT-qPCR method was used to detect the expression level and diversity of NLRP3 and NLRP1 in PBMC and granulocyte mRNA in patients with RA, and detect the mRNA expression of downstream factor IL-1α. The correlation between RA and the expression of NLRP3 and NLRP1 was analyzed. Normal 30 cases were set as control group.@*RESULTS@#Expression levels of NLRP1, and caspase-1 mRNA in PBMC of RA group were significantly lower than those of control group (P0.05); NLRP3, caspase-1, and ASC mRNA expression in granulocyte of RA patients were significantly lower than those in control group (P0.05); NLRP1, IL-1α mRNA expression level had a negative correlation with anti-rheumatoid factor antibody (P=0.033 2, 0.034 0).@*CONCLUSIONS@#NLRP3 and NLRP1 inflammasomes signaling pathways are involved in RA inflammatory reaction process as protective factors, and play an important role in RA inflammatory mechanisms.