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OBJECTIVE: To observe the effects of acupuncture at "antihypertensive acupoint prescription" on endothelial active factors and related autonomic neurotransmitters in spontaneous hypertension rats, and explore the vascular regulation and central regulation mechanisms of acupuncture for anti-hypertension. METHODS: Thirty SPF grade male spontaneous hypertension rats were randomly divided into a model group (15 rats) and an acupuncture group (15 rats). Besides, 15 Wistar Kyoto rats were collected as a blank control group (normal group). In the acupuncture group, acupuncture was delivered at the "antihypertensive acupoint prescription" (bilateral "Renying" [ST 9], "Quchi" [LI 11], "Zusanli" [ST 36], "Taichong" [LR 3] and "Neiguan" [PC 6]), with needles retained for 30 min, once daily. The duration of intervention was 28 days. Every week, using the the irritation scale, the sign of sympathetic irritation was evaluated dynamically. The arterial blood pressure of the rats tail was determined, using non-invasive blood pressure measurement system. ELISA was adopted to detect the levels of calcitonin gene-related peptide (CGRP), nitric oxide (NO), endothelin-1 (ET-1), neuropeptide Y (NPY) in the serum. DAB chromogenic in situ hybridization (CISH) was provided to detect the mRNA expression of endothelial nitric oxide synthase (eNOS) in the internal carotid artery and the arcuate nucleus (ARC), and that of CGRP in the paraventricular nucleus posterior (PVP) and the ventrolateral medulla (VLM). Liquid chromatography-mass spectrometry (LC-MS) was used to detect the levels of epinephrine (E) and norepinephrine (NE) in the paraventricular nucleus anterior (PVA). RESULTS: Compared with the normal group, the irritation scores, systolic blood pressure and diastolic blood pressure were increased at each time point in the model group (P<0.05). When compared with the model group, the irritation scores after the intervention for 3 and 4 weeks, and systolic and diastolic blood pressure after intervention for 2, 3 and 4 weeks were reduced in the acupuncture group (P<0.05). In comparison with the normal group, the serum CGRP and NO levels of the rats were decreased (P<0.05), and the serum ET-1 and NPY levels, as well as E and EN levels in PVA were increased (P<0.05) in the model group. The levels of serum CGRP and NO were elevated (P<0.05), and the serum ET-1 and NPY levels, as well as E and EN levels of PVA were reduced (P<0.05) in the acupuncture group when compared with those of the model group. In the model group, the media of internal carotid artery exhibited thickening and remodeling, while the neuron volume in ARC was small. In the acupuncture group, every layer of internal carotid artery was acceptably arranged, and the parvicellular neuron of ARC was moderate in volume. For the in situ hybridization of eNOS mRNA for the rats of each group, the smooth muscle cells were predominantly expressed in each layer of the internal carotid artery, whereas the expression of parvicellular neurons was dominated in ARC. In the model group, the large and small neurosecretory cells were distributed sparsely in the nerves of PVP; in the acupuncture group, the cells of these two species were distributed regularly; and there were few species of glial cell in the VLM of either the model group or the acupuncture group. In each group, for the in situ hybridization of CGRP mRNA, the small neurosecretory cells were expressed predominately in the PVP, while, the expression of glial cell nuclei and the cell cytoplasm was dominated in the VLM. Compared with the normal group, the mRNA expression of eNOS in the internal carotid artery and ARC and that of CGRP mRNA in the PVP and VLM was decreased in the model group (P<0.05). In the acupuncture group, when compared with the model group, the mRNA expression of eNOS in the internal carotid artery and ARC and that of CGRP in the PVP and VLM was increased in the acupuncture group (P<0.05). CONCLUSION: Acupuncture at "antihypertensive acupoint prescription" can upregulate the level of vascular relaxing factors, downregulate the level of contracting factors, enhance the response of relaxing factors in targeting blood vessels and regulating the center. The mechanism may be related to the modulation of the sympathetic-adrenergic autonomic neurotransmitters in the paraventricular nucleus in spontaneous hypertension rats.
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Terapia por Acupuntura , Pressão Sanguínea , Peptídeo Relacionado com Gene de Calcitonina , Endotelina-1 , Hipertensão , Neuropeptídeo Y , Óxido Nítrico , Ratos Endogâmicos SHR , Animais , Masculino , Ratos , Hipertensão/terapia , Hipertensão/fisiopatologia , Hipertensão/metabolismo , Humanos , Peptídeo Relacionado com Gene de Calcitonina/metabolismo , Peptídeo Relacionado com Gene de Calcitonina/genética , Endotelina-1/metabolismo , Endotelina-1/sangue , Neuropeptídeo Y/metabolismo , Neuropeptídeo Y/genética , Óxido Nítrico/metabolismo , Neurotransmissores/metabolismo , Ratos Endogâmicos WKY , Óxido Nítrico Sintase Tipo III/metabolismo , Óxido Nítrico Sintase Tipo III/genéticaRESUMO
OBJECTIVES: To observe the effects of electroacupuncture (EA) with "intestinal disease prescription" on the intestinal mucosal barrier and NLRP3 inflammasome in rats with dextran sulfate sodium (DSS)-induced acute ulcerative colitis (UC), and explore the underlying mechanism of EA with "intestinal disease prescription" for the treatment of UC. METHODS: Thirty-two healthy male SPF-grade SD rats were randomly divided into a blank group, a model group, a medication group, and an EA group, with 8 rats in each group. Except for the blank group, the UC model was established by administering 5% DSS solution for 7 days. After modeling, the rats in the medication group were treated with mesalazine suspension (200 mg/kg) by gavage, while the rats in the EA group were treated with acupuncture at bilateral "Tianshu" (ST 25), "Shangjuxu" (ST 37) and "Zhongwan" (CV 12), with the ipsilateral "Tianshu" (ST 25) and "Shangjuxu" (ST 37) connected to the electrodes of the EA instrument, using disperse-dense wave, with a frequency of 10 Hz/50 Hz, and each intervention lasted for 20 minutes. Both interventions were performed once daily for 3 days. The general conditions of rats were observed daily. After intervention, the disease activity index (DAI) score was calculated; colon tissue morphology was observed using HE staining; serum levels of pro-inflammatory cytokines (interleukin [IL]-18, IL-1ß) were measured by ELISA; protein expression of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), and Caspase-1 in colon tissues was detected by Western blot; positive expression of zonula occludens-1 (ZO-1) and Occludin in colon tissues was examined by immunofluorescence. RESULTS: Compared with the blank group, the rats in the model group exhibited poor general conditions, slow body weight gain, shortened colon length (P<0.01), increased DAI score and spleen index (P<0.01), elevated serum IL-18 and IL-1ß levels, and increased protein expression of NLRP3, ASC, and Caspase-1 in colon tissues (P<0.01), along with decreased positive expression of ZO-1 and Occludin in colon tissues (P<0.01). Compared with the model group, the rats in the medication group and the EA group exhibited improved general conditions, accelerated body weight gain, increased colon length (P<0.05), reduced DAI scores and spleen indexes (P<0.05), decreased serum IL-18 and IL-1ß levels, and lower protein expression of NLRP3, ASC and Caspase-1 in colon tissues (P<0.05), as well as increased positive expression of ZO-1 and Occludin in colon tissues (P<0.05). There were no significant differences in the above indexes between the medication group and the EA group (P>0.05). Compared with the blank group, the rats in the model group exhibited disrupted colon mucosal morphology, disordered gland arrangement, and atrophy of crypts, along with significant inflammatory cell infiltration. Compared with the model group, the rats in both the medication group and the EA group showed relatively intact colon mucosal morphology, with restored and improved gland and crypt structures, and reduced inflammatory cell infiltration. CONCLUSIONS: EA with "intestinal disease prescription" has a significant therapeutic effect on DSS-induced UC, possibly by regulating the expression of NLRP3 inflammasome and proteins related to the intestinal mucosal barrier, thereby alleviating symptoms of ulcerative colitis.
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Colite Ulcerativa , Eletroacupuntura , Ratos , Masculino , Animais , Colite Ulcerativa/induzido quimicamente , Colite Ulcerativa/terapia , Inflamassomos/efeitos adversos , Interleucina-18 , Ratos Sprague-Dawley , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Ocludina , Peso Corporal , Caspases/efeitos adversosRESUMO
To study the protective mechanism of acupuncture at "Jiangya Recipe" on chronic ischemic white matter injury in spontaneously hypertensive rats (SHR) and the regulation of Jun N-terminal kinase-N-methyl-D-aspartate receptor (JNK-NMDAR) loop. A hypertensive white matter injury model was established in 46 male SHR rats aged 11 weeks by bilateral common carotid artery tapering (SHR-2VGO). In the SHR sham operation group, only bilateral common carotid arteries were isolated and in the SHR-2VGO modeling group, 36 rats were used for microcoil spring clip implantation to narrow the common carotid arteries and then, after 2 weeks of modeling, rats with impaired motor function were removed, and SHR-2VGO rats with successful final models were randomly divided into the model group, JNK blocking group, and acupuncture group. The sham operation group, model group, and JNK blocking group underwent the same grasping fixation, and the acupuncture group received acupuncture at acupoints "Jiangya Fang" once daily. In the JNK blocker group, an injection cannula was implanted into the lateral ventricle and sp600125 was injected into the lateral ventricle at 4.5 ul/day for 4 weeks. One week after the end of the intervention, white matter lesions were detected by MRI DWI and T2 imaging, and the learning and memory ability of rats was tested by Y-Maze and Passive Avoidance. Myelin density was detected by luxol fast blue (LFB) staining, also axon arrangement, myelin integrity, and thickness of neurons were detected by electron microscopy; neuronal morphology and the number of Nissl bodies in the hippocampus were detected by Nissl staining, dendritic spine density changes were detected by Golgi staining, and JNK, NMDAR1, and N-methyl-D-receptor 2B (NMDAR2B) in DG, CA3 region of hippocampus were detected by immunohistochemistry, protein expression of p-JNK/JNK, p-NMDAR1/NMDAR1, NMDAR2B, GSK3ß protein expression in the fimbria of hippocampus was detected by Western blot. The Y maze test of SHR-2VGO+Acu and SHR-2VGO+ sp600125 group showed that the spontaneous alternating reaction rate increased significantly. At the same time, the incubation period increased significantly and the number of errors decreased significantly in Passive Avoidance. MRI T2WI showed that the white matter high signal of the corpus callosum, internal capsule and hippocampal fimbria in the SHR-2VGO+ sp600125 and SHR-2VGO+Acu groups was significantly lower than that in the SHR-2VGO model group, and the striatum and anterior commissure were not obvious. DWI showed that the SHR-2VGO model group had scattered high signal and limited diffusion movement in both the internal capsule and striatum, but the difference between groups was not obvious. Compared with SHR-2VGO rats, LFB staining of SHR-2VGO + sp600125 and SHR-2VGO +Acu groups showed significant relaxation of myelin porosity in corpus callosum, striatum, inner capsule, anterior commissure and hippocampal fimbria, and electron microscopy showed improved axonal myelin integrity and thickness in corpus callosum region. Also, the number of blue patchy Nissl bodies increased, and the number and complexity of dendritic spines increased significantly in Golgi staining. Immunohistochemical detection showed that JNK levels in DG and CA3 region were increased and NMDAR1 and NMDAR2B levels were decreased in SHR-2VGO+Acu and SHR-2VGO+ sp600125 groups. Meanwhile, protein expressions of GSK3ß, NMDAR1/p-NMDAR1 and NMDAR2B in fimbria of hippocampus were increased, and JNK/P-JNK protein expression decreased. Acupuncture can increase the density and thickness of myelin sheath in white matter areas of corpus callosum, anterior commissure and hippocampal fimbria, increase the number and length of hippocampal neuronal dendrites, and improve hypertensive white matter injury and cognitive decline through JNK-NMDAR pathway.
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OBJECTIVES: To investigate the effect of electroacupuncture(EA) at "Changbing Decoction" on alleviating ulcerative colitis (UC) and regulating the polarization of colonic macrophages in rats, so as to explore its mechanisms underlying improvement of UC. METHODS: Twenty-six male SD rats were randomly divided into 4 groupsï¼normal group(6 rats), model group(8 rats), EA group(6 rats), and western medication group(6 rats). The rat model of UC was established by using 5% dextran sulfate sodium (DSS) solution drinking water for 7 days, followed by drinking 1% DSS solution during treatment period. After 7-day model establishment, EA treatment(10 Hz/50 Hz, 20 min) was applied to "Zhongwan"(CV12), bilateral "Tianshu"(ST25) and "Shangjuxu"(ST37) for 3 d, and rats in the western medication group were given mesalazine suspension(200 mg/kg) by gavage for 3 d. The body weight, spleen weight and colon length of rats were measured. The disease activity index (DAI) score was evaluated. The morphological changes and inflammatory cell infiltration of colon were detected after HE staining and pathological scores were eva-luated. The contents of tumor necrosis factor α(TNF-α), interleukin(IL)-1ß, IL-2 and IL-10 in serum were detected by ELISA. The protein expressions of M1 and M2 macrophage markers nitric oxide synthase (iNOS) and arginase 1(Arg1) were detected by fluorescence double staining and Western blot, respectively. Quantitative real-time PCR was used to detect iNOS and Arg1 mRNA expressions. RESULTS: Compared with the normal group, rats in the model group had increased pathological damage degree and inflammatory cell infiltration in the colon tissue, slowed-down body weight gain, decreased colon length, spleen weight, serum anti-inflammatory factors IL-2 and IL-10 contents, colonic Arg1/CD68 fluorescence positive expression, and Arg1 protein and mRNA expressions(P<0.01, P<0.05), as well as increased DAI scores, colon histopathological scores, contents of serum pro-inflammatory factors TNF-α and IL-1ß, colonic iNOS/CD68 fluorescence positive expression, iNOS protein and mRNA expressions(P<0.01). Compared with the model group, the above indicators were significantly improved in rats of the EA group and the western medication group(P<0.01, P<0.05). CONCLUSIONS: EA of "Changbing Decoction" can improve UC of rats by regulating the polarization of colonic macrophages, inhibiting the generation of M1 macrophages and promoting the generation of M2 macrophages.
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Colite Ulcerativa , Eletroacupuntura , Ratos , Masculino , Animais , Colite Ulcerativa/genética , Colite Ulcerativa/terapia , Interleucina-10/genética , Ratos Sprague-Dawley , Fator de Necrose Tumoral alfa/genética , Interleucina-2 , Macrófagos , RNA Mensageiro , Peso CorporalRESUMO
OBJECTIVE: To observe the effect of electroacupuncture ï¼EAï¼ at "Zhongwan" ï¼CV12ï¼, "Tianshu" ï¼ST25ï¼ and "Shangjuxu" ï¼ST37ï¼ ï¼an acupoint prescription "Changbingfang" for treatment of intestinal disordersï¼ on autophagy and expression of AMPK/mTOR signaling pathway in rats with ulcerative colitis ï¼UCï¼, so as to explore its mechanism underlying improvement of UC. METHODS: Thirty-two male SD rats were randomly divided into control, model, medication and EA groups, with 8 rats in each group. The UC model was established by free drinking of 5% dextran sulfate sodium salt solution for 7 days. EA stimulation ï¼10 Hz/50 Hzï¼ was delivered to CV12, ST25 and ST37 for 20 min, once a day for 3 consecutive days. Rats of the medication group received gavage of mesalazine suspension ï¼200 mg/kgï¼ once a day, 3 times in total. The rats' general conditions were recorded for calculating the disease activity index ï¼DAIï¼ score ï¼0-4 pointsï¼. Histomorphological changes of colon were observed via HE staining. The levels of serum interleukin 6 ï¼IL-6ï¼, tumor necrosis factor-α ï¼TNF-αï¼ and IL-10 were measured by ELISA. The mRNA expressions of LC3B and p62 were tested by fluorescence quantitative PCR. Western blot was used to detect the expression levels of LC3B, p62 and AMPK/mTOR pathway related proteins in colon tissues. RESULTS: Compared with the control group, the DAI score, contents of serum IL-6 and TNF-α, the expression levels of p62 protein and mRNA, ratio of p-mTOR/mTOR were significantly increased ï¼P<0.01ï¼ï¼ while the content of serum IL-10, the expression levels of LC3B mRNA, ratio of LC3Bâ ¡/LC3Bâ and p-AMPK/AMPK were decreased ï¼P<0.01, P<0.05ï¼ in the model group. Relevant to the model group, modeling-induced increases of DAI score, serum IL-6, TNF-α and IL-10 contents, expressions of p62 protein and mRNA, LC3B mRNA, ratio of p-mTOR/mTOR, LC3Bâ ¡/LC3Bâ and p-AMPK/AMPK were reversed in both medication and EA groups ï¼P<0.01, P<0.05ï¼. The effect of EA was apparently superior to that of mesalazine in up-regulating ratio of LC3Bâ ¡/LC3Bâ and p-AMPK/AMPK, p62 mRNA expression ï¼P<0.01, P<0.05ï¼, and in down-regulating ratio of p-mTOR/mTOR ï¼P<0.05ï¼. H.E. staining showed severe damage of the colonic mucosal barrier with infiltration of a large number of inflammatory cells in the model group, which was milder in medication and EA groups. CONCLUSION: EA of acupoint recipe "Changbingfang" can improve the symptoms in UC rats, which may be related to its functions in promoting colonic autophagy, increasing AMPK phosphorylation level, and decreasing mTOR phosphorylation level.
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Colite Ulcerativa , Eletroacupuntura , Masculino , Animais , Ratos , Ratos Sprague-Dawley , Colite Ulcerativa/genética , Colite Ulcerativa/terapia , Proteínas Quinases Ativadas por AMP/genética , Interleucina-10 , Mesalamina , Interleucina-6 , Fator de Necrose Tumoral alfa/genética , Transdução de Sinais , Serina-Treonina Quinases TOR/genética , RNA Mensageiro , AutofagiaRESUMO
Fusarium head blight (FHB) is a destructive wheat disease present throughout the world, and host resistance is an effective and economical strategy used to control FHB. Lack of adequate resistance resource is still a main bottleneck for FHB genetics and wheat breeding research. The synthetic-derived bread wheat line C615, which does not carry the Fhb1 gene, is a promising source of FHB resistance for breeding. A population of 198 recombinant inbred lines (RILs) produced by crossing C615 with the susceptible cultivar Yangmai 13 was evaluated for FHB response using point and spray inoculations. As the disease phenotype is frequently complicated by other agronomic traits, we used both traditional and multivariate conditional QTL mapping approaches to investigate the genetic relationships (at the individual QTL level) between FHB resistance and plant height (PH), spike compactness (SC), and days to flowering (FD). A linkage map was constructed from 3,901 polymorphic SNP markers, which covered 2,549.2 cM. Traditional and conditional QTL mapping analyses found 13 and 22 QTL for FHB, respectively; 10 were identified by both methods. Among these 10, three QTL from C615 were detected in multiple years; these QTL were located on chromosomes 2AL, 2DS, and 2DL. Conditional QTL mapping analysis indicated that, at the QTL level, SC strongly influenced FHB in point inoculation; whereas PH and SC contributed more to FHB than did FD in spray inoculation. The three stable QTL (QFhbs-jaas.2AL, QFhbp-jaas.2DS, and QFhbp-jaas.2DL) for FHB were partly affected by or were independent of the three agronomic traits. The QTL detected in this study improve our understanding of the genetic relationships between FHB response and related traits at the QTL level and provide useful information for marker-assisted selection for the improvement of FHB resistance in breeding.
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Sharp eyespot is a major fungal disease of wheat caused by Rhizoctonia cerealis in cool and humid environments worldwide. In this study, 224 single seed descent derived F13, F14 and F15 recombinant inbred lines (RILs) from the cross between CI12633 (a resistant cultivar) and Yangmai 9 (a susceptible cultivar) were assessed for sharp eyespot resistance (R.cerealis isolate R0301) in field and greenhouse conditions in three growing seasons. Different agronomic characteristics were also evaluated in the field with no disease infection. All the lines were genotyped with the Illumina iSelect 90 K SNP wheat chip and 101 SSR markers. Sharp eyespot resistance was significantly negatively correlated with heading date and tiller angle, and significantly positively correlated with the diameter of the basal first internode and second internode. Five QTL with a likelihood of odds ratio score of higher than 3.0 were detected on chromosomes 2BS, 4BS, 5AL and 5BS, respectively. These identified QTL may be used in future wheat breeding programs through marker assisted selection for developing sharp eyespot resistant cultivars.
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Resistência à Doença/genética , Doenças das Plantas , Polimorfismo de Nucleotídeo Único , Locos de Características Quantitativas , Rhizoctonia , Triticum , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Triticum/genética , Triticum/microbiologiaRESUMO
In order to study the influencing factors on Raman spectroscopy, we research a series of comparative Raman spectroscopy of multi-walled carbon nanotubes (MWCNT) with different tube diameter and length. The results suggest that the G peak and D peak of MWCNT are all red-shifted as compared to that of polycrystalline graphite; In the same conditions, the peak intensity (G peak and D peak) is directly proportional to the diameter of the MWCNT, and inversely proportional to the length of the MWCNT; G peak frequency shift is closely related to the MWCNT diameter and length, which are inversely proportional to the diameter (with identical results of the single-walled carbon nanotube radial breathing modes) and direct proportional to the length. While, the influences of the diameter and length on D peak frequency shift are weak, and future analysis for the reason of this kind of phenomenon is as follows. Subsequently, we investigated the relation between D peak frequency shift and MWCNT aspect ratio, the relationship between G peak frequency shift and aspect ratio is nearly linear increase. Using the same analysis method, we plotted the different graphs of G peak and D peak intensity vs the aspect ratio of MWCNT, respectively. As the expected, the linear degression relation are existent in the two relationships.