Puerarin partly counteracts the inflammatory response after cerebral ischemia/reperfusion via activating the cholinergic anti-inflammatory pathway

Puerarin, a major isoflavonoid derived from the Chinese medical herb radix puerariae （Gegen）, has been reported to inhibit neuronal apoptosis and play an anti-inflammatory role in focal cerebral ischemia model rats. Recent findings regarding stroke pathophysiology have recognized that anti-inflammation is an important target for the treatment of ischemic stroke. The cholinergic anti-inflammatory pathway is a highly robust neural-immune mechanism for inflammation control. This study was to investigate whether activating the cholinergic anti-inflammatory pathway can be involved in the mechanism of inhibiting the inflammatory response during puerarin-induced cerebral ischemia/reperfusion in rats. Results showed that puerarin pretreatment （intravenous injection） re- duced the ischemic infarct volume, improved neurological deficit after cerebral ischemia/reperfusion and decreased the levels of interleukin-1β, interleukin-6 and tumor necrosis factor-a in brain tissue. Pretreatment with puerarin （intravenous injection） attenuated the inflammatory response in rats, which was accompanied by janus-activated kinase 2 （JAK2） and signal transducers and activators of transcription 3 （STAT3） activation and nuclear factor kappa B （NF-KB） inhibition. These observa- tions were inhibited by the alpha7 nicotinic acetylcholine receptor （a7nAchR） antagonist a-bungarotoxin （a-BGT）. In addition, puerarin pretreatment increased the expression of a7nAchR mRNA in ischemic cerebral tissue. These data demonstrate that puerarin pretreatment strongly protects the brain against cerebral ischemia/reperfusion injury and inhibits the inflammatory re- sponse. Our results also indicated that the anti-inflammatory effect of puerarin may partly be medi- ated through the activation of the cholinergic anti-inflammatory pathway.

Berberine Relieves Insulin Resistance via the Cholinergic Anti-inflammatory Pathway in HepG2 Cells

Berberine（BBR） is an isoquinoline alkaloid extracted from Rhizoma coptidis and has been used for treating type 2 diabetes mellitus（T2DM） in China. The development of T2 DM is often associated with insulin resistance and impaired glucose uptake in peripheral tissues. In this study, we examined whether BBR attenuated glucose uptake dysfunction through the cholinergic anti-inflammatory pathway in Hep G2 cells. Cellular glucose uptake, quantified by the 2-[N-（7-Nitrobenz-2-oxa-1,3-diazol-4-yl）-amino]-2-deoxy-D-glucose（2-NBDG）, was inhibited by 21% after Hep G2 cells were incubated with insulin（10-6 mol/L） for 36 h. Meanwhile, the expression of alpha7 nicotinic acetylcholine receptor（α7n ACh R） protein was reduced without the change of acetylcholinesterase（ACh E） activity. The level of interleukin-6（IL-6） in the culture supernatant, the ratio of phosphorylated I-kappa-B kinase-β（IKKβ） Ser181/IKKβ and the expression of nuclear factor-kappa B（NF-κB） p65 protein were also increased. However, the treatment with BBR enhanced the glucose uptake, increased the expression of α7n ACh R protein and inhibited ACh E activity. These changes were also accompanied with the decrease of the ratio of p IKKβ Ser181/IKKβ, NF-κB p65 expression and IL-6 level. Taken together, these results suggest that BBR could enhance glucose uptake, and relieve insulin resistance and inflammation in Hep G2 cells. The mechanism may be related to the cholinergic anti-inflammatory pathway and the inhibition of ACh E activity.

Electro-acupuncture regulates the cholinergic anti-inflammatory pathway in a rat model of chronic obstructive pulmonary disease

Objective： Acupuncture has a definite therapeutic effect on chronic obstructive pulmonary disease （COPD）, and the cholinergic anti-inflammatory pathway （CAP） has been shown to be involved in regula- tion of inflammation. In this study, we investigated whether electro-acupuncture （EA） affects the CAP in COPD,Methods： Sprague-Dawley rats were induced into COPD through exposure to cigarette smoke combined with lipopolysaccharide. EA treatment was applied at Zusanli （ST36） and Feishu （BL13） points for 30 min/d for 7 d. Seventy-two rats were randomly divided into six study groups, including normal, normal ＋ EA, normal ＋ α-bungarotoxin （α-BGT） （the antagonist of the nicotinic acetylcholine receptor α7 subunit （α7nAChR）） ＋ EA, COPD, COPD ＋ EA, and COPD ＋ α-BGT ＋ EA. Lung function, pathology and vagus nerve discharge were tested. The levels of acetylcholine （ACh）, acetylcholinesterase （ACHE）, interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-ct） in bronchoalveolar lavage fluid （BALF） and lung tissue were measured by enzyme-linked immunosorbent assay. The mRNA and protein expression and immunoreac- tivity of α7nAChR and its postreceptor inflammation signal pathway, including janus kinase 2 （JAK2）, sig- nal transducers and activators of transcription 3 （STAT3）, nuclear factor-KB （NF-KB）, were observed by quantitative reverse transcription-polymerase chain reaction, Western blot and immunohistochemistry. Results： Compared with normal rats, there were a significant decline in lung function and discharge of the vagus nerve （P 〈 0.01）, a marked sign of lung inflammation and an increase of ACh, ACHE, IL-6 and TNF-α level in BALF or lung tissue （P 〈 0.05, P 〈 0.01 ） and higher expression of 0t7nAChR, JAK2, STAT3 and NF-αB （P 〈 0.05, P 〈 0.01） in the COPD rats. In rats receiving EA, the lung function and vagal discharge were enhanced （P 〈 0.01 ）, lung inflammation was improved and the levels of ACh, ACHE, IL-6 and TNF-α were decreased （P 〈 0.01）. Further, the expression of α7nAChR, JAK2, STAT3 and NF-κB was downregulated （P 〈 0.05, P 〈 0.01 ）. However, the above effects of EA were blocked in rats injected with α-BCT （P 〈 0.01 ）. Conclusion： EA treatment can reduce the lung inflammatory response and improve lung function in COPD, which may be related to its involvement in the regulation of CAP.

Dexmedetomidine attenuates inflammation and pancreatic injury in a rat model of experimental severe acute pancreatitis via cholinergic anti-inflammatory pathway

Background:Excessive inflammatory responses play a critical role in the development of severe acute pancreatitis(SAP),and controlling such inflammation is vital for managing this often fatal disease.Dexmedetomidine has been reported to possess protective properties in inflammatory diseases.Therefore,this study aimed to investigate whether dexmedetomidine pre-treatment exerts an anti-inflammatory effect in rats with SAP induced by sodium taurocholate,and if so,to determine the potential mechanism.Methods:SAP was induced with sodium taurocholate.Rats received an intraperitoneal injection of dexmedetomidine 30 min before sodium taurocholate administration.α-bungarotoxin,a selective alpha-7 nicotinic acetylcholine receptor(α7nAchR)antagonist,was injected intra-peritoneally 30 min before dexmedetomidine administration.The role of the vagus nerve was evaluated by performing unilateral cervical vagotomy before the administration of dexmedetomidine.Efferent discharge of the vagal nerve was recorded by the BL-420F Data Acquisition&Analysis System.Six hours after onset,serum pro-inflammatory cytokine(tumor necrosis factorα[TNF-α]and interleukin 6[IL-6])levels and amylase levels were determined using an enzyme-linked immunosorbent assay and an automated biochemical analyzer,respectively.Histopathological changes in the pancreas were observed after hematoxylin and eosin staining and scored according to Schmidt criteria.Results:Pre-treatment with dexmedetomidine significantly decreased serum levels of TNF-α,IL-6,and amylase,strongly alleviating pathological pancreatic injury in the rat model of SAP(TNF-α:174.2±30.2 vs.256.1±42.4 pg/ml;IL-6:293.3±46.8 vs.421.7±48.3 pg/ml;amylase:2102.3±165.3 vs.3186.4±245.2 U/L).However,the anti-inflammatory and pancreatic protective effects were abolished after vagotomy or pre-administration ofα-bungarotoxin.Dexmedetomidine also significantly increased the discharge frequency and amplitude of the cervical vagus nerve in the SAP rat model(discharge frequency:456.8±50.3 vs.332.4±25.1 Hz;discharge amplitude:33.4±5.3 vs.20.5±2.9μV).Conclusions:Dexmedetomidine administration attenuated the systemic inflammatory response and local pancreatic injury caused by SAP in rats through the cholinergic anti-inflammatory pathway involving vagus-andα7nAChR-dependent mechanisms.

Comparative study of anti-inflammatory effects of different processed products through the COX-2/PGE2 signaling pathway: based on network pharmacology and molecular docking

Background:Radix Aconiti Lateralis Preparata(Fu-zi)is a traditional Chinese medicinal herb,which has been widely used in the clinic and has potent anti-inflammatory activities.we aimed to explore the mechanisms of extract containing alkaloids from different Fu-zi Processed Products(FPP)in treating inflammation,especially rheumatoid arthritis(RA).Methods:Firstly,using network pharmacology technology,the ingredients,and targets of Fu-zi were obtained by searching and screening,the targets involving RA were acquired,the intersection targets were constructed a"component-target-pathway"network.A comprehensive investigation was conducted on the anti-rheumatoid arthritis mechanisms of 5 FPPs in lipopolysaccharide(LPS)induced RAW264.7 cells,which serve as a model for RA.The production of NO and inflammatory cytokines were measured by ELISA kit.Quantitative Real-time PCR(qRT-PCR)was utilized to measure the mRNA levels.COX-2/PGE2 signaling pathway-associated proteins were determined by western blot.Results:According to a network pharmacological study,16 chemical components and 43 common targets were found in Fu-zi and 6 key targets including PTGS2 were closely related to the mechanism of Fu-zi in treating RA.The in vitro study revealed that the levels of NO,TNF-α,and IL-1βwere substantially decreased by the 5 FPPs.The 5 FPPs significantly suppressed the expression of proteins COX-2,iNOS,and NF-κB,with particularly notable effects observed for PFZ and XFZ.Conclusion:Altogether,these results demonstrated that the 5 PPS containing alkaloids have a good anti-RA-related inflammatory effect,and the mechanism may be related to COX-2/PGE2 signaling pathway,particularly,Fu-zi prepared utilizing a traditional Chinese technique.

Impairment of the nerve growth factor pathway driving amyloid accumulation in cholinergic neurons the incipit of the Alzheimer's disease story?

The current idea behind brain pathology is that disease is initiated by mild disturbances of common physiological processes. Overtime, the disruption of the neuronal homeostasis will determine irreversible degeneration and neuronal apoptosis. This could be also true in the case of nerve growth factor （NGF） al- terations in sporadic Alzheimer＇s disease （AD）, an age-related pathology characterized by cholinergic loss, amyloid plaques and neurofibrillary tangles. In fact, the pathway activated by NGF, a key neurotrophin for the metabolism of basal forebrain cholinergic neurons （BFCN）, is one of the first homeostatic systems affected in prodromal AD. NGF signaling dysfunctions have been thought for decades to occur in AD late stages, as a mere consequence of amyloid-driven disruption of the retrograde axonal transport of neuro- trophins to BFCN. Nowadays, a wealth of knowledge is potentially opening a new scenario： NGF signaling impairment occurs at the onset of AD and correlates better than amyloid load with cognitive decline. The recent acceleration in the characterization of anatomical, functional and molecular profiles of early AD is aimed at maximizing the efficacy of existing treatments and setting novel therapies. Accordingly, the elucidation of the molecular events underlying APP metabolism regulation by the NGF pathway in the sep- to-hippocampal system is crucial for the identification of new target molecules to slow and eventually halt mild cognitive impairment （MCI） and its progression toward AD.

MicroRNA-124 mediates the cholinergic anti-inflammatory action through inhibiting the production of pro-inflammatory cytokines

The vagus nerve can control inflammatory response through a ＇ cholinergic anti-inflammatory pathway＇, which is mediated by the α7-nicotinic acetylcholine receptor （α7nAChR） on macrophages. However, the intracel- lular mechanisms that link α7nAChR activation and pro-inflammatory cytokine production remain not well under- stood. In this study, we found that miR-124 is upregulated by cholinergic agonists in LPS-exposed cells and mice. Utilizing miR-124 mimic and siRNA knockdown, we demonstrated that miR-124 is a critical mediator for the cho- linergic anti-inflammatory action. Furthermore, our data indicated that miR-124 modulates LPS-induced cytokine production by targeting signal transducer and activator of transcription 3 （STAT3） to decrease IL-6 production and TNF-α converting enzyme （TACE） to reduce TNF-ot release. These results also indicate that miR-124 is a potential therapeutic target for the treatment of inflammatory diseases.

Involvement of M3 Cholinergic Receptor Signal Transduction Pathway in Regulation of the Expression of Chemokine MOB-1, MCP-1 Genes in Pancreatic Acinar Cells

Whether M3 cholinergic receptor signal transduction pathway is involved in regulation of the activation of NF-κB and the expression of chemokine MOB-1, MCP-1genes in pancreatic acinar cells was investigated. Rat pancreatic acinar cells were isolated, cultured and treated with carbachol, atropine and PDTC in vitro. The MOB-1 and MCP-1 mRNA expression was detected by using RT-PCR. The activation of NF-κB was monitored by using electrophoretic mobility shift assay. The results showed that as compared with control group, M3 cholinergic receptor agonist (10 -3 mol/L, 10 -4 mol/L carbachol) could induce a concentration-dependent and time-dependent increase in the expression of MOB-1, MCP-1 mRNA in pancreatic acinar cells. After treatment with 10 -3 mol/L carbachol for 2 h, the expression of MOB-1, MCP-1 mRNA was strongest. The activity of NF-κB in pancreatic acinar cells was significantly increased (P<0.01) after treated with M3 cholinergic receptor agonist (10 -3 mol/L carbachol) in vitro for 30 min. Either M3 cholinergic receptor antagonist (10 -5 mol/L atropine) or NF-κB inhibitor (10 -2 mol/L PDTC) could obviously inhibit the activation of NF-κB and the chemokine MOB-1, MCP-1 mRNA expression induced by carbachol (P<0.05). This inhibitory effect was significantly increased by atropine plus PDTC (P<0.01). The results of these studies indicated that M3 cholinergic receptor signal transduction pathway was likely involved in regulation of the expression of chemokine MOB-1 and MCP-1genes in pancreatic acinar cells in vitro through the activation of NF-κB.

Exploitation of the nicotinic anti-inflammatory pathway for the treatment of epithelial inflammatory diseases

Discoveries in the first few years of the 21st century have led to an understanding of important interactions between the nervous system and the inflammatory response at the molecular level, most notably the acetylcholine （ACh）- triggered,α7-nicotinic acetylcholine receptor （α7nAChR）- dependent nicotinic antinflammatory pathway. Studies using the α7nAChR agonist, nicotine, for the treatment of mucosal inflammation have been undertaken but the efficacy of nicotine as a treatment for inflammatory bowel diseases remains debatable. Further understanding of the nicotinic anti-inflammatory pathway and other endogenous anti-inflammatory mechanisms is required in order to develop refined and specific therapeutic strategies for the treatment of a number of inflammatory diseases and conditions, including periodontitis, psoriasis, sarcoidosis, and ulcerative colitis.

Selenium-enriched oolong tea(Camellia sinensis)extract exerts anti-inflammatory potential via targeting NF-κB and MAPK pathways in macrophages

Both tea polyphenols and selenium(Se)have been suggested to exert the health benefits via the regulatory capacities of chronic inflammation,which make Se-enriched oolong tea a promising beverage as an anti-inflammatory diet.The aim of this study is to investigate the anti-inflammatory effects of Se-enriched oolong tea extract(Se-TE)and underlying mechanism in lipopolysaccharide(LPS)-induced RAW264.7 cells.Se-TE treatments(50 and 150μg/m L)significantly suppressed the over-production of nitric oxide(NO)and prostaglandin E2(PGE2)in LPS-stimulated macrophages via downregulating the expression of nitric oxide synthase(i NOS)and cyclooxygenase-2(COX-2).Moreover,Se-TEs also effectively inhibited the productions of inflammatory cytokines,such as tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-1β(IL-1β).Furthermore,Se-TE could block mitogen-activated protein kinase(MAPK)and nuclear factor-kappa B(NF-κB)signaling pathways through the inhibition of the phosphorylation of key proteins(IκB-α,p65,p38,ERK,and JNK)and the translocation of the p65 subunit into the nucleus.Collectively,our results indicated that Se-TE may have the potential to be used as a novel food ingredient for the development of various anti-inflammatory foods and the treatment and prevention of chronic inflammation-related diseases.

Electroacupuncture targeting the immune system to alleviate sepsis

Sepsis is a life-threatening inflammatory syndrome with high morbidity and mortality rates.However,options for sepsis are still limited to general treatment in intensive care units(ICUs),and effective therapies that improve sepsis survival are required.Immune disturbances play a vital role in the pathology of sepsis and are associated with protracted inflammation,susceptibility to infections,and death.Therefore,many investigators have focused on the potential benefits of immunomodulation therapy for sepsis.Electroacupuncture(EA)has been practiced in clinics for many years and has shown advantages in treating infectious diseases.Over the last few decades,our understanding of the efficacy and mechanisms of EA in sepsis has undergone considerable developments.We searched the literature regarding“CNKI,Wan Fang Data,VIP Database,PubMed,and Ingenta Connect”from 2010 to 2023,using the keywords“sepsis”“septic”and“electroacupuncture”and 336 sources were searched.Finally,we included 82 studies that targeted the immune system to determine EA’s anti-inflammatory and immunomodulatory effects on sepsis.In this review,we found that EA has clinical benefits in relieving septic inflammation,improving immune function,and attenuating related multi-organ injury through several mechanisms,such as activation of the cholinergic anti-inflammatory pathway(CAP),vagaladrenal axis,inhibition of the nuclear factor Kappa-B(NF-κB)signaling pathway,signal transducers and activators of transcription(STAT)signaling pathway,and improvement of immune cell function.Therefore,EA may be a promising complementary therapy for sepsis treatment.We also expect these data will contribute to further studies on EA in sepsis.

Synthesis and Characterization of Naproxen-Salicylate Derivatives as Potential Dual-Targeted Inhibitors of Dihydrofolate Reductase

Dihydrofolate reductase (DHFR) is an enzyme that catalyzes the reduction of dihydrofolate (DHF) to tetrahydrofolate (THF). Chemotherapy drugs such as methotrexate help to slow the progression of cancer by limiting the ability of dividing cells to make nucleotides by competitively inhibiting DHFR. Nonsteroidal anti-inflammatory drugs (NSAIDs) have been previously reported to exhibit competitive inhibition of DHFR, in addition to their primary action on cyclooxygenase enzymes. This interaction interferes with the enzymatic reduction of dihydrofolate to tetrahydrofolate, thereby impeding the folate metabolism pathway essential for nucleotide synthesis and cell proliferation. This activity stems from their structural resemblance to the p-aminobenzoyl-l-glutamate (pABG) moiety of folate, a substrate of DHFR. It has been established that NSAIDs containing a salicylate group (which has structural similarities to pABG), such as diflunisal, exhibit stronger DHFR-binding activity. In this study, we synthesized salicylate derivatives of naproxen with the aim of exploring their potential as inhibitors of DHFR. The interactions between these derivatives and human DHFR were characterized using a combination of biochemical, biophysical, and structural methods. Through polyacrylamide gel electrophoresis (PAGE) analysis, enzymatic assays, and quantitative ELISA, we investigated the binding affinity and inhibitory potency of the synthesized salicylate derivatives towards DHFR. The findings of this study suggest the potential of salicylate derivatives of naproxen as promising candidates for the inhibition of DHFR, thereby offering novel therapeutic opportunities for modulating the inflammatory process through multiple pathways. Further optimization of these derivatives could lead to the development of more efficacious dual-targeted analogs with enhanced therapeutic benefits.

Effect of the Monocyte Locomotion Inhibitory Factor (MLIF) a Natural Anti-Inflammatory Produced by E. Histolytica on Apoptosis in Human CD4 + T Lymphocytes

Apoptosis prevents the extravasation of intracellular material and the subsequent inflammatory response. Currently, it is not known whether Monocyte Locomotion Inhibitor Factor (MLIF), an anti-inflammatory pentapeptide, induces programmed cell death. We evaluated the effect of MLIF on extrinsic and intrinsic apoptosis pathways human CD4 + T lymphocytes. Cells were cultured for 24 h in RPMI-1640 medium alone (control) or in RPMI medium containing MLIF alone, PMA alone, PMA + MLIF or actinomycin D. Annexin V/propidium iodide-stained cells in early apoptosis showed that cells treated with MLIF or PMA + MLIF were not significantly different from control cells in medium;in contrast, cells treated with PMA or PMA + MLIF demonstrated significant differences from the control in delayed apoptosis. Cytochrome c and caspase 3 levels in cells treated with MLIF showed no significant differences from control cells, however, compared to the control, cells treated with PMA and PMA + MLIF exhibited a significant increase in cytochrome c and caspase 3 levels, which demonstrates that this weak induction of cell death is regulated by the intrinsic pathway of apoptosis. The Fas receptor was not detected in cell culture with any of the treatments employed, suggesting that the extrinsic pathway of apoptosis is not involved. The MLIF per se does not induce apoptosis in human CD4 + T lymphocytes;there may be an additional effect of PMA + MLIF producing the low levels of cell death recorded in the late apoptosis phase. MLIF acts as a natural, biological anti-inflammatory compound produced in axenic cultures of E. histolytica that does not cause apoptosis or elicit an immune response due to its small size, which could make it a strong candidate for future clinical applications.

The role of microglia in depression

According to studies,neuroinflammation is increasingly being linked to the development of major depressive disorder(MDD).In response to inflammatory stimuli,brain microglia,which are immune cells,can change into reactive states.Because of this,microglia play an essentiall role in the early stages of neuroinflammation.Experiments have shown that microglia are able to detect infected or damaged cells,which then activates a cytotoxic response that further exacerbates the harm to brain cells.It has been proven that microglia are quite good at recognizing infections and damaged cells.Microglia,on the other hand,have been found to respond in a number of ways to injury and may even help regenerate damaged tissues.Chronic activation of microglia has been observed in persons with MDD.Deficits in neuroplasticity have been linked to depression,and recent studies show that this may be related to changes in microglia shape and function brought on by either excessive inflammatory activity or the natural aging process.Changing the phenotype of microglia by regulation of inflammatory pathways may be necessary for harnessing neuroinflammation in MDD.Recent research has linked several microglial phenotypes to individual metabolic pathways,showing that energy metabolism plays a pivotal role in coordinating microglial activity.In this study,we investigate whether or not traditional pro-inflammatory,anti-inflammatory,and metabolic pathways in microglia can be used as novel therapeutic routes for regulating neuroinflammation in brain diseases.The focus of this essay is on MDD,although we will also discuss related mental health issues.

Anti-inflammatory germacrane-type sesquiterpene lactones from Vernonia sylvatica

Nine new germacranolides,sylvaticalides A−H(1-9),and three known analogues(10-12)were isolated from the aeri-al part of Vernonia sylvatica.Their structures were established using comprehensive spectroscopic analysis,including high-resolution electrospray ionization mass spectroscopy(HR-ESI-MS)and 1D and 2D nuclear magnetic resonance(NMR)spectra.Their absolute configurations were determined by X-ray diffraction experiments.The anti-inflammatory activities of all isolated compounds were as-sessed by evaluating their inhibitory effects on the nuclear factor kappa B(NF-κB)pathway,which was activated by lipopolysacchar-ide(LPS)-stimulated human THP1-Dual cells,and the interferon-stimulated gene(ISG)pathway,activated by STING agonist MSA-2 in the same cell model.Compounds 1,2 and 6 showed inhibitory effects on the NF-κB and ISG signaling pathways,with IC_(50)values ranging from 4.12 to 10.57μmol·L^(−1).

Effect of Platelet-rich Plasma in Stimulating Macrophage Transformation into M2 Type under AMPK Signaling Pathway

Objective:To explore the effect of platelet-rich plasma(RPR)in stimulating the transformation of pro-inflammatory(M1)macrophages into antiinflammatory(M2)under the adenosine-monophosphate-dependent protein kinase(AMPK)signaling pathway.Methods:Rat peritoneal macrophages(RAW264.7)were cultured and randomly divided into 8 groups:blank control group,LPS group,RPR group A,RPR group B,LPS+RPR(12 h)group,LPS+RPR(24 h)group A,LPS+RPR(24 h)group B,LPS+RPR(24 h)group C.RPR was prepared based on blood donors.The expressions of AMPK signaling pathway-related proteins(AMPK,ULK1,m TOR)and macrophage markers(i NOS,Arg-1)in the blank control group,LPS group,LPS+RPR(12 h)group and LPS+RPR(24 h)group were observed and compared.The expressions of macrophage markers in LPS+RPR(24 h)B and C groups were compared,and the expressions of AMPK and TGF-βin RPR A and B groups were compared.Results:The gray values of AMPK and ULK1 in LPS cells decreased significantly,while those in LPS+RPR(12 h)and LPS+RPR(24 h)A cells increased significantly.The gray values of AMPK and ULK1 in LPS+RPR(24 h)A cells were higher than those in LPS+RPR(12 h)cells(P<0.05).The m TOR gray value of LPS cells was significantly higher than that of LPS+RPR(24 h)A cells,and the m TOR gray value of LPS+RPR(24 h)A cells was significantly lower than that of LPS+RPR(12 h)cells(P<0.05).The expression of i NOS in LPS cells was significantly decreased,the expression of i NOS in LPS+RPR(12 h)and LPS+RPR(24 h)cells was significantly increased,and the expression of i NOS in LPS+RPR(24 h)cells was higher than that in LPS+RPR(12 h)cells(P<0.05).The expression of Arg-1 in LPS cells was significantly decreased,the expression of Arg-1 in LPS+RPR(12 h)and LPS+RPR(24 h)A cells was significantly increased,and the expression of Arg-1 in LPS+RPR(24 h)A cells was higher than that in LPS+RPR(12 h)cells(P<0.05).The i NOS expression level of LPS+PRP(24 h)C cells was significantly higher than that of LPS+PRP(24 h)B cells,and the Arg-1 expression level was significantly lower than that of LPS+PRP(24 h)B cells(P<0.05).The gray values of AMPK and TGF-βin PRP B cells were significantly lower than those in PRP A cells(P<0.05).Conclusion:RPR can stimulate macrophage transformation from M1 to M2 by up-regulating AMPK signaling pathway.

Septic encephalopathy: When cytokines interact with acetylcholine in the brain

Sepsis-associated encephalopathy(SAE) is a brain dysfunction that occurs secondary to infection in the bo characterized by alteration of consciousness, ranging from delirium to coma, seizure or focal neurological signs. S involves a number of mechanisms, including neuroinflammation, in which the interaction between cytokines a acetylcholine results in neuronal loss and alterations in cholinergic signaling. Moreover, the interaction also occurs the periphery, accelerating a type of immunosuppressive state. Although its diagnosis is not specific in biochemis and imaging tests, it could potentiate severe outcomes, including increased mortality, cognitive decline, progress immunosuppression, cholinergic anti-inflammatory deficiency, and even metabolic and hydroelectrolyte imbalan Therefore, the bilateral communication between SAE and the multiple peripheral organs and especially the immu system should be emphasized in sepsis management.

Therapeutic potential of α7 nicotinic receptor agonists to regulate neuroinflammation in neurodegenerative diseases

Neurodegenerative diseases, such as Alzheimer＇s, Parkinson＇s and Huntington＇s diseases, are all character- ized by a component of innate immunity called neuroinflammation. Neuronal loss and neuroinflammation are two phenomena closely linked. Hence, the neuroinflammation is a relevant target for the management of the neurodegenerative diseases given that, to date, there is no treatment to stop neuronal loss. Several studies have investigated the potential effects of activators of alpha 7 nicotinic acetylcholine receptors in animal models of neurodegenerative diseases. These receptors are widely distributed in the central nervous system. After activation, they seem to mediate the cholinergic anti-inflammatory pathway in the brain. This anti-inflammatory pathway, first described in periphery, regulates activation of microglial cells considered as the resident macrophage population of the central nervous system. In this article, we shortly review the agonists of the alpha 7 nicotinic acetylcholine receptors that have been evaluated in vivo and we focused on the selective positive allosteric modulators of these receptors. These compounds represent a key element to enhance receptor activity only in the presence of the endogenous agonist.

Alpha-7 nicotinic acetylcholine receptor agonist treatment in a rat model of Huntington's disease and involvement of heme oxygenase-1

Neuroinflammation is a common element involved in the pathophysiology of neurodegenerative diseases.We recently reported that repeated alpha-7 nicotinic acetylcholine receptor（α7 n ACh R） activations by a potent agonist such as PHA 543613 in quinolinic acid-injured rats exhibited protective effects on neurons.To further investigate the underlying mechanism,we established rat models of early-stage Huntington＇s disease by injection of quinolinic acid into the right striatum and then intraperitoneally injected 12 mg/kg PHA 543613 or sterile water,twice a day during 4 days.Western blot assay results showed that the expression of heme oxygenase-1（HO-1）,the key component of the cholinergic anti-inflammatory pathway,in the right striatum of rat models of Huntington＇s disease subjected to intraperitoneal injection of PHA 543613 for 4 days was significantly increased compared to the control rats receiving intraperitoneal injection of sterile water,and that the increase in HO-1 expression was independent of change in α7 n ACh R expression.These findings suggest that HO-1 expression is unrelated to α7 n ACh R density and the increase in HO-1 expression likely contributes to α7 n ACh R activation-related neuroprotective effect in early-stage Huntington＇s disease.