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OBJECTIVES: We researched the effect and mechanism of acupuncture treatment for visceral pain in rats with diarrhea-predominant irritable bowel syndrome (IBS-D). METHODS: We set up a rat model of IBS-D with chemical and chronic- and acute-pressure stimulations. Then, the IBS-D rats were treated with acupuncture or 5-BDBD, and the therapeutic efficacy of acupuncture in IBS-D rats was assessed by means of the Bristol scale, diarrhea index, abdominal withdrawal reflex (AWR) score, mast cell count and histologic staining. RESULTS: Acupuncture significantly decreased clinical symptoms in IBS-D rats after a 14 day-treatment. Furthermore, significant down-regulation of P2X4, OX42, BDNF (brain-derived neurotrophic factor) and IRF-5 (interferon regulatory factor 5) expressions were observed in the IBS-D rats, along with the decreased inflammatory factors [interleukin 1 beta (IL-1ß), tumor necrosis factor alpha (TNF-α), and interleukin 6 (IL-6)], chemokines [monocyte chemoattractant protein-1 (MCP-1), regulated on activation, normal T cell expressed and secreted (RANTES), and C-X-C motif chemokine ligand 1 (CXCL1)], and neurotransmitters [substance P (SP), 5-hydroxytryptamine (5-HT), and calcitonin gene-related peptide (CGRP)]. 5-BDBD treatment had a similar effect on IBS-D rats. CONCLUSIONS: Acupuncture can effectively alleviate abdominal pain by decreasing visceral hypersensitivity and controlling the expression of P2X4 and spinal microglial inflammation in IBS rats.
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BACKGROUND: Huang-Pu-Tong-Qiao formula (HPTQ), a traditional Chinese medicine (TCM) formula used to improve cognitive impairment. However, the underlying neuroprotective mechanism of HPTQ treated for diabetic cognitive dysfunction (DCD) remains unclear. The purpose of this study was to investigate the neuroprotective mechanism of HPTQ in DCD mice based on molecular docking. METHODS: To investigate the neuroprotective effect of HPTQ in DCD, the Morris water maze (MWM), novel object recognition (NOR) test was used to detect the learning and memory changes of mice; hematoxylin-eosin (HE) staining was used to investigate the damage of hippocampal neurons; the western blot (WB) was used to examine the level of brain-derived neurotrophic factor (BDNF) of hippocampus. To investigate the neuroprotective mechanism of HPTQ in DCD, molecular docking was used to predict the possible target proteins of different active components in HPTQ and then the WB was used to verify the expression of key target proteins in the hippocampus of mice. RESULTS: HPTQ improved the learning and memory ability, hippocampal neuron damage, and the level of BDNF in the hippocampus of the DCD model treated with HFD/STZ for 12 weeks. Besides, the results of molecular docking showed that the main chemical components of HPTQ could be well combined with the targets of Bcl-2-associated X (Bax) and B-cell lymphoma2 (Bcl-2) and caspase-3. The levels of Bax/Bcl-2 protein ratio and caspase-3 increased in the DCD model while the HPTQ inhibited it. In addition, HPTQ restored DCD-induced decline of p-CREB, BDNF, TrkB, and p-Akt in the hippocampus. CONCLUSIONS: These data indicated that HPTQ ameliorates the hippocampus apoptosis in diabetic cognitive dysfunction mice by activating CREB/BDNF/TrkB signaling pathway.
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OBJECTIVE: To observe the effect of electroacupuncture (EA) stimulation of "Zusanli"(ST36)on gastric function (food consumption and gastric emptying rate) and excitability of hippocampal glutamatergic neurons in mice, so as to explore its mechanism underlying enhancing gastrointestinal function. METHODS: The present study includes 2 parts. 1) C57BL/6 mice were randomly divided into normal and EA groups (n=12 in each group). EA (2 Hz/15 Hz, 1-3 mA) was applied to bilateral ST36 for 20 min, once daily for 7 days. In each group, 6 mice were used to measure the food consumption and gastric emptying rate, and the other 6 mice used to detect the hippocampal glutamate secretion content by using in vivo microdialysis and high performance liquid chromatography. 2) Thirty CaMKIIα-Cre mice received microinjection of a recombinant adeno-associated viral vector containing inhibitory designer receptor exclusively activated by a designer drug (DREADD, AAV-DIO-hM4Di-eYFP) into the hippocampus. Twenty-one days later, 3 mice were selected to observe the expression of eYFP-labeled hM4Di by immunohistochemistry, and 15 mice employed to observe the electrical activities of hM4Di-eYFP positive neurons exposed in chemogenetic activating drug Clozapine N-oxide (CNO) perfusion conditions (n=3) and without CNO in the recording chamber (n=6 in the control and EA groups) by using whole cell patch clamp. The rest 12 CaMKII-Cre mice were equally randomized into AAV-DIO-hM4Di-eYFP+CNO group and AAV-DIO-hM4Di-eYFP+CNO+EA group, and CNO was given by intraperitoneal injection for observing the effect of EA on gastric function. RESULTS: 1) In C57BL/6 mice, compared with the normal group, the food consumption, gastric emptying rate, and the glutamate content in the hippocampus were obviously increased in the EA group (P<0.01). 2) In CaMKIIα-Cre mice, the hM4Di-eYFP positive neurons distributing in the hippocampus showed an obvious increase of firing rates in the EA group relevant to the control group (P<0.01), and a hyperdepolarization potential after application of CNO. No significant changes were found between the AAV-DIO-hM4Di-eYFP+CNO and AAV-DIO-hM4Di-eYFP+CNO+EA groups in the food consumption and gastric emptying rate (P>0.05), suggesting an elimination of EA effect after acute DREADD-mediated activation of the CaMKIIα-positive hippocampal excitatory neurons. CONCLUSION: EA at ST36 can promote food intake and gastric emptying in normal mice but not in CaMKIIα-Cre mice with activated hippocampal hM4Di receptorsï¼suggesting a contribution of the CaMKIIα-positive hippocampal excitatory neurons (glutamatergic neurons in particular) to the enhanced gastrointestinal function of EA at ST36.
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Eletroacupuntura , Animais , Esvaziamento Gástrico , Hipocampo , Camundongos , Camundongos Endogâmicos C57BL , NeurôniosRESUMO
The symbiont endophytic fungi in tobacco are highly diverse and difficult to classify. Here, we sequenced the genomes of Curvularia trifolii and Leptosphaerulina chartarum isolated from tobacco plants. Finally, 41.68 Mb and 37.95 Mb nuclear genomes were sequenced for C. trifolii and L. chartarum with the scaffold N50, accounting for 638.94 Kb and 284.12 Kb, respectively. Meanwhile, we obtained 68,926 bp and 59,100 bp for their mitochondrial genomes. To more accurately classify C. trifolii and L. chartarum, we extracted seven nuclear genes and 12 mitochondrial genes from these two genomes and their closely related species. The genes were then used for calculation of evolutionary rates and for phylogenetic analysis. Results showed that it was difficult to achieve consistent results using a single gene due to their different evolutionary rates, while the phylogenetic trees obtained by combining datasets showed stable topologies. It is, therefore, more accurate to construct phylogenetic relationships for endophytic fungi based on multi-gene datasets. This study provides new insights into the distribution and characteristics of endophytic fungi in tobacco.
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Ascomicetos/classificação , Ascomicetos/genética , Genoma Fúngico , Genoma Mitocondrial , Genômica , Nicotiana/microbiologia , Filogenia , Ascomicetos/isolamento & purificação , Evolução Molecular , Genômica/métodos , Sequenciamento de Nucleotídeos em Larga Escala , Análise de Sequência de DNARESUMO
Bioactivity-guided isolation of the endophytic fungus Fusarium sambucinum TE-6L residing in Nicotiana tabacum L. led to the discovery of two new angularly prenylated indole alkaloids (PIAs) with pyrano[2,3-g]indole moieties, amoenamide C (1) and sclerotiamide B (2), and four known biosynthetic congeners (3-6). Their structures were determined by comprehensive spectroscopic techniques, electronic circular dichroism (ECD), and X-ray diffraction. Compound 1 containing the bicyclo[2.2.2]diazaoctane core and indoxyl unit is rarely reported. All the compounds were evaluated for their antimicrobial and insecticidal activities. Notably, compounds 1-3 showed potent inhibitory effects against three human- and one plant-pathogenic bacterium, and seven plant-pathogenic fungi. Compounds 2-4 also exhibited remarkable larvicidal activity against first instar larvae of the cotton bollworm Helicoverpa armigera with mortality rates of 70.2%, 83.2%, and 70.5%, respectively. Further toxicity tests on zebrafish embryos were performed to evaluate the potential toxicity of PIAs. Of significance was that compound 3 in particular exhibited the highest activities but the lowest effects on the hatching of embryos among all the compounds. This study provides a basis for understanding developmental toxicity of PIAs exposure to zebrafish embryos, and also indicates the potential environmental risks of other natural compounds exposure in the aquatic ecosystem.
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Anti-Infecciosos/química , Endófitos/química , Fusarium/química , Alcaloides Indólicos/química , Inseticidas/química , Animais , Anti-Infecciosos/metabolismo , Anti-Infecciosos/farmacologia , Bactérias/efeitos dos fármacos , Endófitos/isolamento & purificação , Fungos/efeitos dos fármacos , Fusarium/isolamento & purificação , Alcaloides Indólicos/metabolismo , Alcaloides Indólicos/farmacologia , Inseticidas/metabolismo , Inseticidas/farmacologia , Larva/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Estrutura Molecular , Mariposas/efeitos dos fármacos , Nicotiana/microbiologia , Peixe-Zebra/embriologiaRESUMO
OBJECTIVE: To observe the effects of electroacupuncture (EA) on gastric motility, protooncogene c-fos and hippocampus N-methyl-D-aspartate (NMDA) receptor subunits in rats with functional dyspepsia (FD), and to discuss the molecular mechanism of hippocampal in EA at "Zhongwan" (CV 12) and "Weishu" (BL 21) for gastric motility. METHODS: Eighty-four Sprague Dawley (SD) rats were randomly divided into a normal group, a model group, a Zhongwan group, a Weishu group, an acupoint combination group and a non-acupoint group, 14 rats in each one. Except for the normal group, FD model were established by moderate tail-clipping infuriation method and irregular feeding. The rats in the Zhongwan group, Weishu group, acupoint combination group and non-acupoint group were treated with EA at corresponding acupoints, 20 min per treatment, once a day for 7 days. The rats in the normal group and the model group received no treatment; grabbing and fixation were applied in the model group. The stress transducer was used to record gastric motion waveforms; immunohistochemistry method was used to detect the expression of c-fos in hippocampus; Western blot method was used to detect the expression of NMDA receptor subunits NR1, NR2A and NR2B in hippocampus. RESULTS: Compared with the normal group, the gastric motility range was decreased (P<0.05), and the expression of hippocampus NR2A was reduced and the levels of NR1 and NR2B were increased in the model group (P<0.05), however, hippocampus c-fos expression was not significant different between the normal group and the model group (P>0.05). Compared with the model group, the gastric motility range was increased, the expression of hippocampus c-fos and expression of hippocampus NR2A was increased but expressions of NR1 and NR2B were reduced in the Weishu group, Zhongwan group and acupoint combination group (P<0.05). Besides, the differences of gastric motility range and expression of hippocampus c-fos, NR1, NR2A and NR2B between the model group and the non-acupoint were not significant (P>0.05). Compared with the Zhongwan group and the Weishu group, the gastric motility range was increased, the expression of hippocampus c-fos and NR2A was increased but the expression of NR1 and NR2B was reducedin the acupoint combination group (P<0.05). There was no significant difference in the frequency of gastric motility among the groups (P>0.05). CONCLUSION: EA at "Zhongwan" (CV 12) and "Weishu" (BL 21) could increase gastric motility of FD rats, which is likely to be related with activating hippocampal neurons, upregulating the level of NR2A and downregulating NR1 and NR2B.
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Eletroacupuntura , Animais , Medicamentos de Ervas Chinesas , Hipocampo , Ratos , Ratos Sprague-Dawley , Receptores de N-Metil-D-AspartatoRESUMO
Fungal endophytes are the most ubiquitous plant symbionts on earth and are phylogenetically diverse. Studies on the fungal endophytes in tobacco have shown that they are widely distributed in the leaves, stems, and roots, and play important roles in the composition of the microbial ecosystem of tobacco. Herein, we analyzed and quantified the endophytic fungi of healthy tobacco leaves at the seedling stage (SS), resettling growth stage (RGS), fast-growing stage (FGS), and maturing stage (MS) at three altitudes (600, 1000, and 1300 m). We sequenced the internal transcribed spacer (ITS) region of fungal samples to delimit operational taxonomic units (OTUs) and phylogenetically characterize the communities. The results showed that the numbers of clustering OTUs at SS, RGS, FGS, and MS were 516, 709, 469, and 428, respectively. At the phylum level, species in Ascomycota and Basidiomycota had absolute predominance, representing 97.8% and 2.0% of the total number of species, respectively. We also found the number of fungi at the RGS and FGS stages was higher than those at the other two stages. Additionally, OTU richness was determined by calculating the Observed Species, Shannon, Simpson, Chao1, abundance-based coverage estimator (ACE), Good's coverage and phylogenetic distance (PD)_whole_tree indices based on the total number of species. Our results showed RGS samples had the highest diversity indices. Furthermore, we found that the diversity of fungal communities tended to decrease with increasing altitude. The results from this study indicated that tobacco harbors an abundant and diverse endophytic fungal community, which provides new opportunities for exploring their potential utilization.
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Ascomicetos/genética , Basidiomycota/genética , Variação Genética , Microbiota/fisiologia , Nicotiana/microbiologia , Filogenia , Folhas de Planta/microbiologia , Plântula/microbiologiaRESUMO
Genistein is a kind of isoflavone compoundsï¼ also called phytoestrogensï¼ with clinical effects on cardiovascular diseaseï¼ cancer and postmenopausal-related gynecological diseasesï¼ and also has the potentiality in the prevention and treatment of Alzheimer's disease(AD). In this studyï¼ the protective effect of genistein on Aß25â35-induced PC12 cell injury and effect on CaM-CaMKIV signaling pathway were observed to investigate its mechanism for AD. PC12 cells were cultured in vitro and then the safe concentration of genistein and the modeling concentration and optimal time point of administration of Aß25â35 were screened by MTT assay. After being pretreated with different concentrations of genistein(25ï¼ 50ï¼ 100 µmol·L⻹) on PC12 cellsï¼ the AD model of PC12 cells was induced by Aß25â35. Then the survival rate of cells was detected by MTT assay; morphological change of cells was observed under the inverted microscopeï¼ and apoptosis of cells was assessed by AO/EB fluorescence staining; the neuroprotective effects of genistein on AD cell model were observed and the optimal concentration of genistein was determined. Expressions of mRNA and protein levels of CaMï¼ CaMKKï¼ CaMKIV and tau were detected by qRT-PCR and Western blot assayï¼ respectively. The results showed that as compared with the blank groupï¼ the cell survival rate was decreased; the cell damage and apoptosis were increased; and the expressions of mRNA and protein levels of CaMï¼ CaMKKï¼ CaMKIV and tau were increased in AD model group. Genistein could significantly improve the cell survival rateï¼ reduce the cell damage and apoptosis of AD cell modelï¼ and significantly down-regulate the expressions of mRNA and protein levels of CaMï¼ CaMKKï¼ CaMKIV and tau of AD cell model. These results indicated that genistein has obviously neuroprotective effect on the AD cell model induced by Aß25â35ï¼ and the mechanism may be related to the down-regulation of CaM-CaMKIV signaling pathway and Tau protein expression.
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Peptídeos beta-Amiloides , Proteína Quinase Tipo 4 Dependente de Cálcio-Calmodulina/metabolismo , Calmodulina/metabolismo , Genisteína/farmacologia , Fragmentos de Peptídeos , Substâncias Protetoras/farmacologia , Transdução de Sinais/efeitos dos fármacos , Animais , Apoptose , Sobrevivência Celular , Células PC12 , RatosRESUMO
Diabetes has become a global public health problem that seriously threatens human health. Traditional Chinese medicine, the characteristics of the role of multiple targets, has a unique advantage in the prevention and treatment of diabetes mellitus and its complications. Astragaloside-â £ (AS-â £), one of the main activities of Astragalus membranaceus, has a series of pharmacological effects including improvement in the function of endothelial cells and neovascularization, anti-inflammatory, antioxidant, regulating energy metabolism, protectionnervous, anti-cancer and so on. In this paper, AS-â £ to prevent and treat diabetes and its complications has been reviewed, which has effect on lowering blood sugar, lowering blood pressure, improving insulin resistance, inhibiting inflammatory reaction and oxidative stress. Additionally, it also can improve the diabetic animal and cell model of diabetic vascular disease, diabetic nephropathy, diabetic retinopathy, diabetic peripheral neuropathy, diabetic cardiomyopathy and other pathological damages. AS-â £ may be a potential active substance for the treatment of diabetes and its complications.
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Astragalus propinquus/química , Diabetes Mellitus/tratamento farmacológico , Medicamentos de Ervas Chinesas/farmacologia , Saponinas/farmacologia , Triterpenos/farmacologia , Animais , Complicações do Diabetes/tratamento farmacológico , Angiopatias Diabéticas/tratamento farmacológico , Nefropatias Diabéticas/tratamento farmacológico , Neuropatias Diabéticas/tratamento farmacológico , Humanos , Medicina Tradicional ChinesaRESUMO
AIM: To study the neural mechanism by which electroacupuncture (EA) at RN12 (Zhongwan) and BL21 (Weishu) regulates gastric motility. METHODS: One hundred and forty-four adult Sprague Dawley rats were studied in four separate experiments. Intragastric pressure was measured using custom-made rubber balloons, and extracellular neuron firing activity, which is sensitive to gastric distention in the dorsal vagal complex (DVC), was recorded by an electrophysiological technique. The expression levels of c-fos, motilin (MTL) and gastrin (GAS) in the paraventricular hypothalamic nucleus (PVN) were assayed by immunohistochemistry, and the expression levels of motilin receptor (MTL-R) and gastrin receptor (GAS-R) in both the PVN and the gastric antrum were assayed by western blotting. RESULTS: EA at RN12 + BL21 (gastric Shu and Mu points), BL21 (gastric Back-Shu point), RN12 (gastric Front-Mu point), resulted in increased neuron-activating frequency in the DVC (2.08 ± 0.050, 1.17 ± 0.023, 1.55 ± 0.079 vs 0.75 ± 0.046, P < 0.001) compared with a model group. The expression of c-fos (36.24 ± 1.67, 29.41 ± 2.55, 31.79 ± 3.00 vs 5.73 ± 2.18, P < 0.001), MTL (22.48 ± 2.66, 20.76 ± 2.41, 19.17 ± 1.71 vs 11.68 ± 2.52, P < 0.001), GAS (24.99 ± 2.95, 21.69 ± 3.24, 23.03 ± 3.09 vs 12.53 ± 2.15, P < 0.001), MTL-R (1.39 ± 0.05, 1.22 ± 0.05, 1.17 ± 0.12 vs 0.84 ± 0.06, P < 0.001), and GAS-R (1.07 ± 0.07, 0.91 ± 0.06, 0.78 ± 0.05 vs 0.45 ± 0.04, P < 0.001) increased in the PVN after EA compared with the model group. The expression of MTL-R (1.46 ± 0.14, 1.26 ± 0.11, 0.99 ± 0.07 vs 0.65 ± 0.03, P < 0.001), and GAS-R (1.63 ± 0.11, 1.26 ± 0.16, 1.13 ± 0.02 vs 0.80 ± 0.11, P < 0.001) increased in the gastric antrum after EA compared with the model group. Damaging the PVN resulted in reduced intragastric pressure (13.67 ± 3.72 vs 4.27 ± 1.48, P < 0.001). These data demonstrate that the signals induced by EA stimulation of acupoints RN12 and BL21 are detectable in the DVC and the PVN, and increase the levels of gastrointestinal hormones and their receptors in the PVN and gastric antrum to regulate gastric motility. CONCLUSION: EA at RN12 and BL21 regulates gastric motility, which may be achieved through the PVN-DVC-vagus-gastric neural pathway.
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Pontos de Acupuntura , Eletroacupuntura/métodos , Esvaziamento Gástrico , Núcleo Hipotalâmico Paraventricular/fisiologia , Núcleo Solitário/fisiologia , Estômago/inervação , Nervo Vago/fisiologia , Potenciais de Ação , Animais , Gastrinas/metabolismo , Masculino , Mecanotransdução Celular , Motilina/metabolismo , Vias Neurais/fisiologia , Núcleo Hipotalâmico Paraventricular/metabolismo , Pressão , Proteínas Proto-Oncogênicas c-fos/metabolismo , Ratos Sprague-Dawley , Receptor de Colecistocinina B/metabolismo , Receptores dos Hormônios Gastrointestinais/metabolismo , Receptores de Neuropeptídeos/metabolismo , Núcleo Solitário/metabolismo , Nervo Vago/metabolismoRESUMO
AIM: To investigate the effects and mechanisms of vasoactive intestinal peptide (VIP) and nitric oxide (NO) in the modulation of electroacupucture (EA) on gastric motility in restrained-cold stressed rats. METHODS: An animal model of gastric motility disorder was established by restrained-cold stress. Gastric myoelectric activities were recorded by electrogastroenterography (EGG). VIP and NO concentrations in plasma and gastric mucosal and bulb tissues were detected by radioimmunoassay (RIA). VIP expression in the gastric walls was assayed using avidin-biotin-peroxidase complex (ABC) and image analysis. RESULTS: In cold restrained stressed rats, EGG was disordered and irregular. The frequency and amplitude of gastric motility were higher than that in control group (P < 0.01). VIP and NO contents of plasma, gastric mucosal and bulb tissues were obviously decreased (P < 0.01). Following EA at "Zusanli" (ST36), the frequency and amplitude of gastric motility were obviously lowered (P < 0.01), while the levels of VIP and NO in plasma, gastric mucosal and bulb tissues increased strikingly (P < 0.01, P < 0.05) and expression of VIP in antral smooth muscle was elevated significantly (P < 0.01) in comparison with those of model group. CONCLUSION: VIP and NO participate in the modulatory effect of EA on gastric motility. EA at "Zusanli" acupoint (ST36) can improve gastric motility of the stressed rats by increasing the levels of VIP and NO.