[Mechanism of Liangfu Pills in treatment of functional dyspepsia: based on network pharmacology and experimental verification].

This study aims to explore the potential mechanism of Liangfu Pills in the treatment of functional dyspepsia(FD) based on network pharmacology and molecular docking, and verify the mechanism by animal experiment. The active components of Liangfu Pills were screened from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP), and the targets of Liangfu Pills were predicted by SwissTargetPrediction. The targets of FD were retrieved from GeneCards. On this basis, the common targets of the disease and the pills were yielded and the protein interaction was retrieved based on STRING. The core targets were screened out, followed by Gene Oncology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis with DAVID. Finally, molecular docking was carried out with the help of AutoDock Tools to predict the binding degree between the effective components of Liangfu Pills and core targets. A total of 19 active components of Liangfu Pills and 591 FD-related targets were screened out by network pharmacology, of which 253 were common targets of the disease and the prescription. Liangfu Pills was mainly involved in the biological processes of response to drug, negative regulation of transcription, positive regulation of apoptotic process, and cell surface receptor signaling pathway, and the KEGG pathways of hypoxia-inducible factor-1(HIF-1) signaling pathway, serotonergic synapse, tumor necrosis factor(TNF) signaling pathway, cyclic adenosine monophosphate(cAMP) signaling pathway, calcium signal pathway, and inflammatory mediator regulation of transient receptor potential(TRP) channels. The results of molecular docking showed that the key active components of Liangfu Pills had certain binding activity to the targets mitogen-activated protein kinase 1(MAPK1), protein kinase B(AKT1), transient receptor potential cation channel subfamily V member 1(TRPV1), 5-hydroxytryptamine receptor 1 A(HTR1 A), and 5-hydroxytryptamine receptor 2 A(HTR2 A). FD was induced in rats, and then Liangfu Pills was given to FD rats for 7 days. The results showed that Liangfu Pills could significantly relieve the symptoms of FD rats, significantly increase the expression of 5-hydroxytryptamine(5-HT), and down-regulate the expression of TRPV1. Through network pharmacology, molecular docking, and experimental verification, this study proved that Liangfu Pills improved FD through multiple components and multiple targets. The result lays a basis for further research on the mechanism and clinical application of Liangfu Pills in the treatment of FD.

Tetrandrine Alleviates Nociception in a Rat Model of Migraine via Suppressing S100B and p-ERK Activation in Satellite Glial Cells of the Trigeminal Ganglia.

Sensitization and activation of the trigeminal ganglia have been implicated in the pathology of migraine. Satellite glial cells (SGCs), a specialized type of glial cells that ensheathe trigeminal neurons, may be critical for peripheral nociceptive sensitization. Tetrandrine (TET), an alkaloid extracted from a traditional Chinese herb, exerts an inhibitory effect on glial activation in vitro and has been used in various neurologic diseases. The current study investigated the effect of TET on nitroglycerin (NTG)-induced trigeminal sensitization and examined potential signaling pathways related to SGC activation in the model of migraine. We measured trigeminal nociceptive thresholds using electronic von Frey rigid tips before and after NTG injection in control rats and rats pretreated with TET, while expression and subcellular location of the inflammatory mediators S100B and activated phosphorylation extracellular signal-regulated kinase (p-ERK) were measured using real-time quantitative polymerase chain reaction, Western blotting, and double immunofluorescence staining. Pretreatment with TET caused a dose-dependent reversal of the trigeminal nociceptive hypersensitivity induced by NTG. In addition, TET pretreatment blocked the activation of S100B and p-ERK in trigeminal ganglion SGCs of NTG-treated rats. Reduced p-ERK activity can suppress the inflammation that leads to hyperexcitability of trigeminal ganglion neurons. Administration of TET may therefore be a safe and effective therapeutic treatment for the hyperalgesic symptoms of migraine.

PTEN inhibition preserves trigeminal nucleus caudalis neuron activation through tyrosine phosphorylation of the NR2B subunit at Tyr1472 of the NMDA receptor in a rat model of recurrent migraine.

OBJECTIVE: Activation of the trigeminal nucleus caudalis is believed to be involved in the pathomechanism of migraine. Evidence suggests that N-methyl-d-aspartate receptor subtype 2B tyrosine phosphorylation, originating from the trigeminal nucleus caudalis neuron dysfunction, might be a triggering mechanism for recurrent migraine. Phosphatase and tensin homolog is thought to have a neuroprotective effect in various neurologic diseases by regulating N-methyl-d-aspartate receptor subtype 2B or tyrosine phosphorylation. Thus, the aim of this study was to explore whether the recombinant adenovirus AdR-siPTEN attenuates neuron activation in the trigeminal nucleus caudalis in a rat model of recurrent migraine. METHODS: Adenovirus-expressing siPTEN or RFP was independently injected into the spinal trigeminal nucleus of the rat model suffering from recurrent migraine by inflammatory soup stimulation the superior sagittal sinus of rats. Seven days later, tactile sensory testing was performed to detect the tactile threshold. Immunofluorescence, Immunohistochemistry, and western blot assay were done to measure PTEN, NR2B, NR2B-pTyr1472, and c-Fos levels in the trigeminal nucleus caudalis of recurrent migraine rats. RESULTS: A significant increase (p < 0.05) in neuron c-Fos content, an indicator of neuron activation, was detected in the trigeminal nucleus caudalis in a rat model of recurrent migraine. However, neuron activation in the trigeminal nucleus caudalis was attenuated by pretreatment with AdR-siPTEN. Moreover, the attenuated effect was potentially mediated by tyrosine phosphorylation of the NR2B-p1472 tyrosine site in the trigeminal nucleus caudalis, as seen in rat brain slices. CONCLUSION: These results suggest that, phosphatase and tensin homolog might be a novel and promising candidate for future treatment or prophylaxis of recurrent migraine by attenuating neuron activation in the trigeminal nucleus caudalis.

Neuroprotective effects of tetrandrine against vascular dementia.

Tetrandrine is one of the major active ingredients in Menispermaceae Stephania tetrandra S. Moore, and has specific therapeutic effects in ischemic cerebrovascular disease. Its use in vascular dementia has not been studied fully. Here, we investigated whether tetrandrine would improve behavioral and cellular impairments in a two-vessel occlusion rat model of chronic vascular dementia. Eight weeks after model establishment, rats were injected intraperitoneally with 10 or 30 mg/kg tetrandrine every other day for 4 weeks. Behavioral assessment in the Morris water maze showed that model rats had longer escape latencies in training trials, and spent less time swimming in the target quadrant in probe trials, than sham-operated rats. However, rats that had received tetrandrine showed shorter escape latencies and longer target quadrant swimming time than untreated model rats. Hematoxylin-eosin and Nissl staining revealed less neuronal necrosis and pathological damage, and more living cells, in the hippocampus of rats treated with tetrandrine than in untreated model rats. Western blot assay showed that interleukin-1ß expression, and phosphorylation of the N-methyl-D-aspartate 2B receptor at tyrosine 1472, were lower in model rats that received tetrandrine than in those that did not. The present findings suggest that tetrandrine may be neuroprotective in chronic vascular dementia by reducing interleukin-1ß expression, N-methyl-D-aspartate receptor 2B phosphorylation at tyrosine 1472, and neuronal necrosis.

Replication of migraine GWAS susceptibility loci in Chinese Han population.

BACKGROUND: Recent genome-wide association studies (GWAS) have identified 3 genetic variants that are strongly associated with migraine in Europeans. The effect of these risk variants in other populations is unknown. To further replicate the GWAS findings, we investigated the 3 variants rs2651899 (1p36.32, PRDM16), rs10166942 (2q37.1, TRPM8), and rs11172113 (12q13.3, LRP1) for their association with migraine in the Chinese Han population. METHODS: We performed a case-control association study. Genomic DNA was collected from 608 unrelated individuals, including 304 migraineurs (41 migraine with aura and 263 migraine without aura) and 304 healthy controls. Genotyping of single nucleotide polymorphisms (SNPs) was performed by ligase detection reaction method. RESULTS: We identified the minor allele of rs2651899 located in PRDM16 to be associated with migraine (P = .005, odds ratio = 1.382, 95% confidence interval = 1.100-1.736), the association remain significant after Bonferroni correction. For the other 2 SNPs (rs10166942 and rs11172113), no statistically significant differences were observed in the allele/genotype frequencies between cases and controls. None of the 3 SNP was associated with specific migraine features. CONCLUSION: Our study confirmed the association of PRDM16 to migraine susceptibility in the Chinese Han population. The results also indicated that replication studies of previous GWAS findings across populations is of importance to validate these associations and to gain a better understanding of migraine susceptibility of potential genetic heterogeneity between populations. Further work is necessary to understand the functional mechanisms underlying these variants identified by GWAS.

Preventive effects of AdR-siPTEN through the regulation of NMDA receptor NR2B subunit in trigeminal ganglia of migraine rats.

OBJECTIVE: Migraine is a refractory disease that is due to neuronal hyperexcitability, and has high incidence, mortality, and disability rates. The N-methyl-D-aspartate receptor 2B (NR2B) subunit has been found to play an important role in the pathogenesis of migraine. There is evidence suggesting that a tumor suppressor phosphatase and tensin homolog (PTEN) can confer a neuroprotective effect on cerebral ischemic injury by regulating NR2B levels. However, the role of PTEN in migraines is still unclear. This study aimed to define whether PTEN is involved in the pathogenesis of migraine through modulating NR2B, nitric oxide synthase (NOS), and nitric oxide (NO) in the trigeminal ganglia of rats with glyceryl trinitrate-induced migraine. METHODS: Adenovirus-expressing siPTEN or RFP was independently injected into the Sp5 (spinal trigeminal nucleus) of rats suffering from migraines. Seven days later, tactile sensory testing was performed to detect the tactile threshold. Immunofluorescence assay, western blot assay, RT-PCR, and biochemical examination were done to measure PTEN, NR2B, NOS, and NO levels in the trigeminal ganglia of migraine rats. RESULTS: NR2B, NOS, and NO levels significantly (P<0.05) decreased in the trigeminal ganglia of migraine-affected rats pretreated with adenovirus-expressing siPTEN. CONCLUSION: These results suggest that PTEN in trigeminal ganglia is implicated in the pathogenesis of migraine, and PTEN may be a novel and promising candidate for future treatment and/or prevention of migraine via regulating NR2B and decreasing NO production in trigeminal ganglia.