Experimental study on the regulation effect of Qiaoqin Qingfei agent on cough variant asthma based on airway neurogenic inflammation

Objective:To investigate the mechanism of regulation of airway neurogenic inflammation by Qiaoqin Qingfei agent in rats with cough variant asthma(CVA).Methods:48 SD rats were randomly divided into blank group,model group,montelukast sodium group(1.05 mg/kg)and high,medium and low dose groups(26,13,6.5 g/kg),with 8 rats in each group.The rat CVA model was established by the method of ovalbumin(OVA)combined with aluminum hydroxide(Al(OH)3)sensitization and repeated stimulation.From the second day of sensitization,the rat CVA model was given by gavage for 28 days.The pathological changes of lung tissue were observed under microscope by HE staining.The content changes of nerve growth factor(NGF)and substance P(SP)in alveolar lavage fluid(BALF)were determined by double-antibody sandwich ABC-ELISA,and the protein expression levels of NGF and SP in lung tissue were detected by immunohistochemistry.Results:Pathological findings showed significant inflammatory manifestations in the model group,and the inflammatory infiltration in the high-dose,medium-dose and low-dose groups of Qiaoqin Qingfei agent and montelukast sodium groups were alleviated to varying degrees.Compared with blank group,the protein expression levels of NGF and SP in lung tissue of model group were significantly increased(P<0.01).Compared with model group,the protein expression levels of NGF and SP in lung tissue and the contents of NGF and SP in alveolar lavage fluid in high-dose,medium-dose and low-dose groups and montelukast sodium group were significantly decreased(P<0.05).Conclusion:Qiaoqin Qingfei agent may reduce airway inflammation and relieve cough variant asthma by regulating the protein expression levels of NGF and SP in airway neurogenic inflammation.

NEUROGENIC INFLAMMATION EVOKED BY STIMULATION OF THE ACUPOINT THROUGH DRR

The purpose of this study is to investigate whether stimulation of the acupoint evoked the neurogenic inflammation distributed along the meridian through DRR (dorsal root reflex). We used 11 rats whose ventral roots below L 1 were cut off and the spinal cords were transected between T 13 and L 1. After intravenous injection of Evans blue liquid and continuous electrostimulation of the right Chengshan acupoint of the rat, Evans blue extravasation distributed along the Bladder Meridian of Foot Taiyang was observed on the skin of lower back and both hindlimbs. By means of measuring the amount of Evans blue, it was shown that the amount of Evans blue spot in the skin with the extravasation was obviously different from that in the skin of 1 0 mm apart from (P<0 01), also obviously different from that in the skin of the upper back and from that in the skin of the control group (P<0 01), also whown that the amount of Evans blue in the bladder, ureter and germen was obviously different from that in the control group (P<0 01). These results indicate that the stimulation of the acupoint may produce the neurogenic inflammation of ipsilateral and contralateral skin up to several adjacent spinal segments as well as may produce the neurogenic inflammation of ipsilateral and contralateral visceral organs related to the Bladder Meridian through DRRs.

Efficacy of active compounds of Chanqin granules on airway neurogenic inflammation induced by PM2.5 in vivo

OBJECTIVE:To investigate the efficacy of active compounds of Chanqin(CQ)granules on PM2.5-induced airway neurogenic inflammation in vivo,and to elucidate the underlying mechanisms of action.METHODS:The Traditional Chinese Medicine systems pharmacology(TCMSP)database was searched,and the results were combined with oral bioavailability and drug analysis to identify the compounds in CQ granules.The pharmacophore modeling approach was used to predict the compound targets,and the diseases corresponding to the targets were obtained by searching the therapeutic target database(TTD),pharmacogenomics knowledgebase(Pharm GKB)and Drug Bank databases.Cytoscape software was used to construct the network pharmacological charts for Component-Target and Target-Disease interactions of the CQ granules.Then,the mechanisms of action and effectiveness of CQ granules for the treatment of PM2.5-induced airway neurogenic inflammation were analyzed.RESULTS:A total of 195 compounds and 171 targets were obtained from the analyses.A total of569 corresponding diseases were identified for these targets.Component-target and target-disease networks were constructed.The possible mechanisms and effective components in CQ granules for treating airway neurogenic inflammation were analyzed.Quercetin,kaempferol and isorhamnetin,beta-sitosterol and sitosterol,which are typically found in the formulation,have extensive pharmacological activities,including anti-inflammatory,antioxidant and antiviral actions and neuroprotective properties.Among these targets,androgen receptor,estrogen receptor,prostaglandin G/H synthase 2,and inducible nitric oxide synthase play important pathological roles,including the induction of neurogenic inflammation.CQ granules may have therapeutic effectiveness for numerous diseases in addition to respiratory diseases,including neoplasms,digestive system diseases,cardiovascular diseases,respiratory tract diseases and nervous system diseases.In vivo,CQ granules are effective in treating pulmonary inflammation and downregulate neuropeptides in the bronchoalveolar lavage fluid after PM2.5 exposure.CQ granules significantly decreased the levels of neurokinin A,neurokinin B and calcitonin gene-related peptide in the lung and dorsal root ganglia.CQ also significantly suppressed the upregulation of p-extracellular regulated protein kinase 1/2 and p-methyl ethyl ketone 1/2 induced by PM2.5 exposure.CONCLUSION:CQ granules have potential for thetreatment of neurogenic inflammation induced by PM2.5 in vivo,and the mechanism might involve downregulation of neuropeptides in the BALF,lung and dorsal root ganglia.

Pathogenesis of Parkinson＇s disease： oxidative stress, environmental impact factors and inflammatory processes

Current hypothesis of neuronal degeneration in Parkinson＇s disease （PD） have been proposed, including formation of free radicals and oxidative stress, mitochondrial dysfunction, excitotoxicity, trophic factor deficiency, inflammatory processes, genetic factors, environmental impact factors, toxic action of nitric oxide, apoptosis, and so on. This review mainly discussed oxidative stress, environmental impact factors, and inflammatory processes in PD.

Pathogenesis of Parkinson’s disease: oxidative stress, environmental impact factors and inflammatory processes

Current hypothesis of neuronal degeneration in Parkinson’s disease (PD) have been proposed, including formation of free radicals and oxidative stress, mitochondrial dysfunction, excitotoxicity, trophic factor deficiency, inflam- matory processes, genetic factors, environmental impact factors, toxic action of nitric oxide, apoptosis, and so on. This review mainly discussed oxidative stress, environmental impact factors, and inflammatory processes in PD.

Effects of TRPA1 activation and inhibition on TRPA1 and CGRP expression in dorsal root ganglion neurons

Transient receptor potential ankyrin 1 （TRPA1） is a key player in pain and neurogenic inflammation, and is localized in nociceptive primary sensory dorsal root ganglion （DRG） neurons. TRPA1 plays a major role in the transmission of nociceptive sensory signals. The generation of neurogenic inflammation appears to involve TRPAl-evoked release of calcitonin gene-related peptide （CGRP）. However, it remains unknown whether TRPA1 or CGRP expression is affected by TRPA 1 activation. Thus, in this study, we examined TRPA 1 and CGRP expression in DRG neurons in vitro after treatment with the TRPA1 activator tbrmaldehyde or the TRPA1 blocker menthol. In addition, we examined the role of extracellular signal-regulated protein kinase 1/2 （ERK1/2） in this process. DRG neurons in culture were exposed to formaldehyde, menthol, the ERK1/2 inhibitor PD98059 ＋ formaldehyde, or PD98059 ＋ menthol. After treatment, real-time polymerase chain reaction, western blot assay and double immunofluorescence labeling were performed to evaluate TRPA1 and CGRP expression in DRG neurons. Formaldehyde elevated mRNA and protein levels of TRPA 1 and CGRP, as well as the proportion of TRPA1- and CGRP-positive neurons. In contrast, menthol reduced TRPA1 and CGRP expression. Furthermore, the effects of lbrmaldehyde, but not menthol, on CGRP expression were blocked by pretreatment with PD98059. PD98059 pretreatment did not affect TRPA1 expression in the presence of formaldehyde or menthol.

A novel large animal model of recurrent migraine established by repeated administration of inflammatory soup into the dura mater of the rhesus monkey

Several animal models of migraine have been established, and those based on trigeminovascular system activation are widely accepted. How- ever, most of these models have been established on lower animals, such as rodents, and involve only a single administration of a noxious stimulus. In this study, an inflammatory soup （10 μL）, consisting of prostaglandin E2 （0.2 mM）, serotonin （2 mM）, bradykinin （2 raM） and histamine （2 raM）, was injected into the dura mater of conscious rhesus monkeys through an indwelling catheter. The infusion started on day 8 and was repeated every 3 days, for a total of six administrations, to induce neurogenic inflammation. We performed behavioral assessments and measured the expression of the oncogene c-fos, neuronal nitric oxide synthase （nNOS） and calcitonin gene related peptide （CGRP） ill the trigeminal system and in multiple brain regions involved in pain processing by immunohistochemical staining. Compared with monkeys in the control group, three of the four animals in the inflammatory soup group displayed decreased motor behaviors, and two showed increased ipsilateral nose and mouth secretions during the stimulus period. Higher expression levels of c-fos, nNOS and CGRP were found in various brain areas of experimental animals compared with controls, including the trigeminal nucleus caudalis, thalamus, hypothalamus, midbrain, pons and other areas involved in pain perception. These results suggest that repeated inflammatory soup stimulation of the dura activates the trigeminovascular system and produces migraine-like pathological changes and abnormal behaviors in conscious rhesus monkeys.

PLASMA EXTRAVASATION IN THE VISCERAL ORGANS CAUSED BY ELECTRICAL STIMULATION OF ACUPOINT "ZUSANLI" IN RATS

Objective To investigate the plasma extravasation of visceral organs caused by electrical stimulation of acupoint under the dorsal root reflex and axon reflex conditions. Methods By the means of measuring the content of Evans blue, this study investigated the plasma extravasation of visceral organs induced by electrical stimulation of acupoint 'Zusanli'(ST 36).Results The Evans blue content in the visceral organs such as liver, spleen, pancreas and the whole gastrointestinal tract in rats increased significantly after electrical stimulation of acupoint 'Zusanli' compared with that of the control group (P<0.01). The Evans blue extravasation in the above visceral organs was blocked by pre-treatment of capsaicin (66 mmol·L -1, 50 μL) into the acupoint. Conclusion The neurogenic inflammation of the visceral organs evoked by electrical stimulation of acupoint was mediated by the capsaicin-sensitive afferent fibers through the dorsal root reflex and axon reflex. It is a new method to study the correlation of meridian-viscera.

New Insights of Neuromedin B and Its Receptor NMBR Involvement in Immunity

Neuromedin B(NMB)is a member of mammalian bombesin-like peptide family and almost all its potent biological actions take place exclusively through its receptor NMBR,which has been identified as a G-protein couple receptor with seven trans-membrane regions.NMB and NMBR represent an interesting signaling axis over which NMB/NMBR perform multiple biological activities,in particularly,they have been highlighted in the regulation of the immunology system.This review aims to provide a brief overview of the recent advancements in our understanding regarding the potential immunological functions of NMB and NMBR in cell proliferation,tumorigenesis,angiogenesis,neurogenic inflammation,anti-influenza A virus(IAV)infections,and briefly discusses the potential importance of these observations.All these implicated that targeting NMBR or developing analogues of NMB may represent a useful strategy for development of new anti-IAV agents.

Electroacupuncture at Sensitized Acupoints Relieves Somatic Referred Pain in Colitis Rats by Inhibiting Sympathetic-Sensory Coupling to Interferewith 5-HT Signaling Pathway

Objective:To investigate whether electroacupuncture(EA) at sensitized acupoints could reduce sympathetic-sensory coupling(SSC) and neurogenic inflammatory response by interfering with 5-hydroxytryptamine(5-HT)ergic neural pathways to relieve colitis and somatic referred pain,and explore the underlying mechanisms.Methods:Rats were treated with 5% dextran sodium sulfate(DSS) solution for 7 days to establish a colitis model.Twelve rats were randomly divided into the control and model groups according to a random number table(n=6).According to the Research on Rat Acupoint Atlas",sensitized acupoints and non-sensitized acupoints were determined.Rats were randomly divided into the control,model,Zusanli-EA(ST 36),Dachangshu-EA(BL 25),and Xinshu(BL 15) groups(n=6),as well as the control,model,EA,and EA+GR113808(a 5-HT inhibitor)groups(n=6).The rats in the control group received no treatment.Acupuncture was administered on 2 days after modeling using the stimulation pavameters:1 mA,2 Hz,for 30 min,with sparse and dense waves,for 14 consecutive days.GR113808 was injected into the tail vein at 5 mg/kg before EA for 10 min for 7 consecutive days.Mechanical sensitivity was assessed with von Frey filaments.Body weight and disease activity index(DAI) scores of rats were determined.Hematoxylin and eosin staining was performed to observe colon histopathology.SSC was analyzed by immunofluorescence staining.Immunohistochemical staining was performed to detect 5-HT and substance P(SP) expressions.The calcitonin gene-related peptide(CGRP) in skin tissue and tyrosine hydroxylase(TH) protein levels in DRG were detected by Western blot.The levels of hyaluronic acid(HA),bradykinin(BK),prostaglandin I2(PGI2) in skin tissue,5-HT,tryptophan hydroxylase 1(TPH1),serotonin transporters(SERT),5-HT 3 receptor(5-HT3R),and 5-HT 4 receptor(5-HT4R) in colon tissue were measured by enzyme-linked immunosorbent assay(ELISA).Results:BL 25 and ST 36 acupoints were determined as sensitized acupoints,and BL 15 acupoint was used as a non-sensitized acupoint.EA at sensitized acupoints improved the DAI score,increased mechanical withdrawal thresholds,and alleviated colonic pathological damage of rats.EA at sensitized acupoints reduced SSC structures and decreased TH and CGRP expression levels(P<0.05).Furthermore,EA at sensitized acupoints reduced BK,PGI2,5-HT,5-HT3R and TPH1 levels,and increased HA,5-HT4R and SERT levels in colitis rats(P<0.05).GR113808 treatment diminished the protective effect of EA at sensitized acupoints in colitis rats(P<0.05).Conclusion:EA at sensitized acupoints alleviated DSS-induced somatic referred pain in colitis rats by interfering with 5-HTergic neural pathway,and reducing SSC inflammatory response.

Implications of Transient Receptor Potential Cation Channels in Migraine Pathophysiology

Migraine is a common and debilitating headache disorder. Although its pathogenesis remains elusive,abnormal trigeminal and central nervous system activity is likely to play an important role. Transient receptor potential(TRP) channels, which transduce noxious stimuli into pain signals, are expressed in trigeminal ganglion neurons and brain regions closely associated with the pathophysiology of migraine. In the trigeminal ganglion,TRP channels co-localize with calcitonin gene-related peptide, a neuropeptide crucially implicated in migraine pathophysiology. Many preclinical and clinical data support the roles of TRP channels in migraine. In particular,activation of TRP cation channel V1 has been shown to regulate calcitonin gene-related peptide release from trigeminal nerves. Intriguingly, several effective antimigraine therapies, including botulinum neurotoxin type A, affect the functions of TRP cation channels. Here, we discuss currently available data regarding the roles of major TRP cation channels in the pathophysiology of migraine and the therapeutic applicability thereof.

Role of cortical spreading depression in the pathophysiology of migraine

A migraine is a recurring neurological disorder characterized by unilateral, intense, and pulsatile headaches. In one-third of migraine patients, the attacks are preceded by a visual aura, such as a slowly-propagating scintillating scotoma. Migraine aura is thought to be a result of the neurovascular phenomenon of cortical spreading depression （SD）, a self-propagating wave of depolarization that spreads across the cerebral cortex. Several animal experiments have demonstrated that cortical SD causes intracranial neurogenic inflammation around the meningeal blood vessels, such as plasma protein extravasation and pro-inflammatory peptide release. Cortical SD has also been reported to activate both peripheral and central trigeminal nociceptive pathways. Although several issues remain to be resolved, recent evidence suggests that cortical SD could be the initial trigger of intracranial neurogenic inflammation, which then contributes to migraine headaches via subsequent activation of trigeminal afferents.