The effects and molecular mechanisms of vitamin D3 on airway inflammation and oxidative stress response in a mouse model of bronchial asthma / 安徽医科大学学报

Objective @#To investigate the role and related molecular mechanisms of vitamin D3 ( VitD3 ) in airway inflammation and oxidative stress response in bronchial asthma mice．@*Methods @#Twenty-eight female C57BL/6 mice were randomly divided into a control group ( Ctrl) and a model group．The model group mice were sensitized using ovalbumin ( OVA) to establish an asthma model，and were further divided into an asthma (Asthma) group， VitD3 treatment (Asthma + VitD3 ) group，and Forkhead Box O1 (FOXO1) inhibitor AS1842856 treatment (Asth- ma + AS) group．Lung resistance (LR) changes were measured in each group of mice．Enzyme-linked immunosor- bent assay (ELISA) was used to detect the levels of tumor necrosis factor-alpha (TNF-α) ，interleukin ( IL) -1 β , and IL-18 in bronchoalveolar lavage fluid ( BALF) ．Western blot was used to determine the expression levels of FOXO1，NOD-like receptor pyrin domain containing 3 (NLRP3) ，Caspase-1，and apoptosis-associated speck-like protein (ASC) in lung tissue. @*Results @#ompared to the Ctrl group mice，LR increased in the Asthma group mice (P＜0. 01) ．Compared to the Asthma group，LR decreased in the Asthma + VitD3 and Asthma + AS group mice (P＜0. 05) ，with no significant difference in LR change between Asthma + VitD3 and Asthma + AS group mice. Compared to the Ctrl group，TNF-α , IL-1 β , and IL-18 levels in BALF，as well as NLRP3，Caspase-1，and ASC protein expression levels in lung tissue，increased in the Asthma，Asthma + VitD3 ，and Asthma + AS group mice (P＜0. 05) ．Compared to the Asthma group，the Asthma + VitD3 and Asthma + AS group mice showed decreased levels of the mentioned inflammatory factors in BALF and reduced protein expression of NLRP3，FOXO1，Caspase- 1，and ASC in lung tissue (P＜0. 05) ．Compared to the Asthma + VitD3 group，the Asthma + AS group showed in- creased FOXO1 protein expression (P ＜0. 05) ，with no statistically significant differences in the other measured indicators．@*Conclusion @#VitD3 can alleviate asthma symptoms induced by OVA in mice，improve the degree of air- way inflammation，and reduce oxidative stress levels．The mechanism may be related to the downregulation of the FOXO1/ NLRP3 axis.

Mechanism of salidroside preventing myocardial fibrosis based on TLR4-mediated pyroptosis pathway / 中国药房

OBJECTIVE To investigate the effects of salidroside （Sal） on myocardial fibrosis and pyroptosis and its potential mechanism. METHODS The mice were randomly divided into control group， model group and Sal low-dose， medium-dose and high-dose groups， with 10 mice in each group. Except for the control group， the mice in other groups were injected subcutaneously with isoproterenol 5 mg/（kg·d）to prepare the myocardial fibrosis model. Since modeling， mice in the Sal low-dose， medium-dose and high-dose groups were given 10， 30 and 50 mg/kg of Sal by intragastric administration every day； control group and model group were given 10 mL/kg of normal saline by intragastric administration every day， for 14 consecutive days. After the last medication， the mice were sacrificed； hematoxylin-eosin staining was used to observe pathological change of myocardial tissue and calculate the diameter of myocardial cell； Masson and Sirius Red staining were used to observe the degree of myocardial fibrosis in mice and calculate the collagen volume fraction （CVF）； quantitative real-time PCR was performed to detect the mRNA expressions of collagen type Ⅰ （Col Ⅰ）， α-smooth muscle actin （α-SMA）， Toll-like receptor 4 （TLR4）， NOD-like receptor pyrin domain containing 3 （NLRP3）， caspase-1 andgasdermin D （GSDMD） in myocardial tissues. The total protein expressions of Col Ⅰ， α-SMA， TLR4， NLRP3，caspase-1 and GSDMD in myocardial tissues and protein-positive cell score were measured by Western blot assay and immunohistochemistry. RESULTS Compared with control group， the myocardial cells in the model group were enlarged， the arrangement of myocardial fibers was disordered， the matrix metabolism was significantly increased， the CVF in myocardial tissue was significantly increased， and the mRNA and protein expression levels of Col Ⅰ， α-SMA， TLR4， NLRP3， caspase-1 and GSDMD were elevated and protein-positive cell score was increased significantly （P＜0.01）. Compared with model group， the myocardial cell morphology was clearer， myocardial fibrosis was alleviated， and the levels of the above indicators in myocardial tissue of Sal medium-dose and high-dose groups had been reversed to varying degrees， especially in Sal high-dose group（P＜0.05 or P＜0.01）. In addition， the Sal low-dose group also reversed some fibrosis and pyroptosis-related indicators to some extent. CONCLUSIONS Sal can significantly prevent the occurrence and development of myocardial fibrosis， and the mechanism of action may be related to the inhibition of TLR4-mediated pyroptosis pathway in myocardial tissue.

Babaodan Alleviates APAP-induced Acute Liver Injury in Mice by Inhibiting NLRP3/Caspase-1 Pathway / 中国实验方剂学杂志

ObjectiveTo explore the effect of Babaodan （BBD） on the NOD-like receptor pyrin domain containing 3/cysteine aspartate-specific protease-3 （NLRP3/Caspase-1） pathway proteins in mice with acetaminophen （APAP）-induced acute liver injury. MethodC57BL/6 mice were randomly grouped， and BBD （75， 150， 300 mg·kg-1， ig） was administered twice a day for three days. After 2 hours of the last administration， the mice were treated with APAP （400 mg·kg-1， ip）， and the eyeballs were removed to collect blood after 14 hours. Then they were sacrificed by cervical dislocation for sample collection. Hematoxylin-eosin （HE） staining was used to observe the morphological changes of liver tissue cells， and biochemical methods were used to detect the activities of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， superoxide dismutase （SOD）， malondialdehyde （MDA） and myeloperoxidase （MPO） in serum of mice in each group. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was performed to determine the mRNA expression of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β） and IL-6， and Western blot was performed to determine the protein expression of cyclooxygenase-2 （COX-2）， inducible nitric oxide synthase （iNOS）， NLRP3， Caspase-1 and IL-18 in the liver of mice. ResultCompared with the conditions in normal group， the hepatic lobule structure of mice in the model group was partially destroyed， and the hepatic sinusoids were dilated. And the expression levels of ALT and AST in serum， the protein levels of NLRP3， Caspase-1， iNOS， IL-18 and COX-2 and the mRNA levels of IL-1β， IL-6 and TNF-α were increased （P<0.05， P<0.01）. Compared with the model group， the administration groups had improvement in liver cell rupture and hepatic sinusoidal compression， and a dose-dependent decrease in the levels of ALT and AST in serum as well as the protein levels of NLRP3， Caspase-1， iNOS， IL-18 and COX-2 and the the mRNA levels of IL-1β， IL-6 and TNF-α in liver tissue （P<0.05， P<0.01）. ConclusionBBD can reduce APAP-induced acute liver injury in mice. The mechanism may be related to anti-oxidative stress， inhibition of NLRP3/Caspase-1 pathway， and decreased expression levels of IL-1β， IL-18， TNF-α and IL-6.

Effect of NLRP3 Inflammasomes on Development of Diabetes Mellitus and Its Complications and Chinese Medicine Intervention via NLRP3： A Review / 中国实验方剂学杂志

Diabetes mellitus （DM） is a metabolic disease mainly characterized by chronic hyperglycemia and has multiple etiologies. The complications of DM， such as coronary atherosclerosis， nephropathy， foot disease and cardiac dysfunction， have high morbidity， disability rate and mortality. DM and its complications have a long course of disease and are easy to relapse， which are difficult to be cured， seriously affecting people's life and health. NOD-like receptor pyrin domain-containing 3 （NLRP3） inflammasome is an important component of inflammatory response and innate immune system. The inflammatory cascade induced by NLRP3 activation is involved in the occurrence and development of DM as well as its complications by releasing inflammatory factors， damaging endothelial cells and affecting metabolic stress. Therefore， as the core of the inflammatory response， NLRP3 may provide a new target for the treatment of DM and its complications. Traditional Chinese medicine plays a key role in the treatment of DM and its complications， and has a regulatory effect on NLRP3. Thus it has become a novel research strategy to prevent and treat DM and its complications via modulating NLRP3. However， at present， there are relatively scattered reports and a lack of systematic review on the role of traditional Chinese medicine in the treatment of DM and its complications from the perspective of NLRP3. As a result， this paper reviewed domestic and foreign literature in recent years and conducted the discussion from two aspects： the influence of NLRP3 on the occurrence and development of DM and its complications， and the progress of traditional Chinese medicine in intervening in DM and its complications through NLRP3. This paper provided reference for the research on the regulation of NLRP3 and a new direction for the treatment of DM and its complications.

Relationship between NLRP3 activation level of inflammasome and the change of cognitive functions in patients with acute ischemic stroke / 中华行为医学与脑科学杂志

Objective:To investigate the relationship between the activation level of Nod-like receptor pyrin domain-containing 3 (NLRP3) inflammasome and the change of cognitive functions in patients with acute ischemic stroke.Methods:A total of 88 patients with acute ischemic stroke in Department of Neurology from October 2018 to July 2020 were selected as case group and 100 healthy physical examinees were selected as control group.Peripheral blood of the case group and the control group was collected, and peripheral blood mononuclear cells (PBMCs) were isolated by centrifugation.Then the NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), Caspase-1 and interleukin-1β (IL-1β) expression were detected by Western blot.The cognitive function of patients with acute ischemic stroke was detected by Montreal Cognitive Assessment (MoCA). The differences in expression levels of NLRP3, ASC, Caspase-1 and IL-1β were compared between the case group and the control group.Pearson correlation analysis was used to analyze the correlation between expression levels of NLRP3, ASC, Caspase-1, IL-1β and MoCA score.Logistic multivariate regression was used to analyze the relationship between expression levels of NLRP3, ASC, caspase-1, IL-1β and the cognitive dysfunction.Results:(1)Western blot results showed that NLRP3, ASC, Caspase-1 and IL-1β expressions in PBMCs cells in the case group were higher than those in the control group (all P<0.05). (2)The expression level of NLRP3 in stroke patients with hypertension, hyperlipidemia, National Institutes of Health Stroke Scale (NIHSS) score ≥ 8 points was significantly higher than that in patients without hypertension, hyperlipidemia and NIHSS score<8 points ( P<0.05); (3)The incidence of cognitive dysfunction in the case group was 34.09% (30/88). The MoCA scores of the cognitive dysfunction group and the non-cognitive dysfunction group were 20 (24, 28) and 27 (26, 28) points respectively, and the difference between the groups was statistically significant ( P<0.05); (4)Pearson correlation analysis showed that NLRP3, ASC, caspase-1 and IL-1β expression in PBMCs cells were negatively correlated with MoCA scores ( r=-0.426, -0.396, -0.417, -0.320 respectively, all P<0.05). (5)Logistic regression analysis showed that hyperlipidemia, NIHSS scores, frontotemporal lobe infarction, and NLRP3 expression were the influencing factors for the occurrence of cognitive dysfunction (all P<0.05). Conclusion:Patients with acute ischemic stroke have high activated NLRP3 inflammasome, and its activation degree is closely related to the condition and the occurrence of cognitive dysfunction after stroke.Targeted inhibition or regulation of NLRP3 inflammasome activation may become a new idea of neuroprotection for acute ischemic stroke.

Recent advance in role of purinergic 2X7 receptor receptor and Nod like receptor pyrin domain containing 3 inflammasome in central nervous system diseases / 中华神经医学杂志

Purinergic 2X7 receptor (P2X7R) and Nod like receptor pyrin domain containing 3 (NLRP3) inflammasome contribute to the inflammatory activation.Great attention has been paid to P2X7R and NLRP3 inflammasome in recent years,especially in the fields of central nervous system.To further elucidate the role of the P2X7R and NLRP3 inflammasome in the central nervous system,we review the latest research progress of composition and physiological functions of P2X7R and NLRP3 inflammasome and how they play their roles in diseases of the central nervous system.

NOD-like receptor pyrin domain containing 3 inflammasome and myocardial ischemia-reperfusion injury / 中南大学学报(医学版)

Myocardial ischemia-reperfusion injury (MI/RI) is an inflammatory cascade process involving the interaction of multiple factors.In recent years,more and more evidence suggests that NOD-like receptor pyrin domain containing 3 (NLRP3) inflammasome,an important component of the innate immune system,is closely associated with the inflammatory damage of MI/RI.Furthermore,blockage of NLRP3 inflammasome or the release of its downstream pro-inflammatory cytokines may provide new therapeutic targets for this disorder.

Effect of electroacupuncture pretreatment on expression of NLRP3 in neurons during cerebral ischemia-reperfusion in rats / 中华麻醉学杂志

Objective To evaluate the effect of electroacupuncture (EA) pretreatment on NODlike receptor pyrin domain-containing 3 (NLRP3) in neurons during cerebral ischemia-reperfusion (I/R) in rats.Methods Fifty-four adult male Sprage-Dawley rats,aged 7 weeks,weighing 250-280 g,were randomly divided into 3 groups (n =18 each) using a random number table:sham operation group (group S),group I/R,and EA pretreatment group (group E).Cerebral I/R was induced by occlusion of the right middle cerebral artery for 90 min using a nylon thread inserted into the internal carotid artery and advanced intracranially to block the blood flow,followed by reperfusion.In group E,the acupoint Baihui was stimulated with an electric stimulator (sparse-dense wave,frequency 2 Hz/15 Hz,intensity ≤ 1 mA) for 30 min once a day for 5 consecutive days,and the model of cerebral I/R was established at 24 h after the last stimulation.At 72 h of reperfusion,neurological function was assessed and scored.The rats were then sacrificed,and their brains were removed for determination of cerebral infarct volume (using TTC staining),expression of NLRP3,caspase-1 and interleukin-1beta (IL-1β) in brain tissues (by Western blot),and expression of NLRP3 protein in neurons (by immunofluorescence histochemistry).The percentage of cerebral infarct volume was calculated.Results Compared with group S,the percentage of cerebral infarct volume and neurological scores were significantly decreased,and the expression of NLRP3,caspase-1 and IL-1β in brain tissues was significantly up-regulated in group I/R (P＜0.05).Compared with group I/R,the percentage of cerebral infarct volume and neurological scores were significantly increased,and the expression of NLRP3,caspase-1 and IL-1β in brain tissues was significantly down-regulated ingroup E (P＜0.05).Conclusion The mechanism by which EA pretreatment reduces inflammatory responses during cerebral I/R injury may be related to down-regulation of NLRP3 expression in neurons in rats.