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ObjectiveTo investigate the mechanism of Magnolia officinalis cortex for constipation-type irritable bowel syndrome（IBS-C） rats before and after sweating. MethodIBS-C rat model was established by gavage of ice water， and rats were randomly divided into the blank group， model group， mosapride group（1 mg·kg-1）， M. officinalis cortex group（10 g·kg-1） and sweated M. officinalis cortex group（10 g·kg-1）. The changes of body weight， fecal number and fecal water content of rats were observed， 16S rRNA sequencing was used to detect the structural changes of fecal intestinal flora in rats， the levels of 5-hydroxytryptamine（5-HT） and substance P（SP） in colonic tissues of rats were determined by enzyme-linked immunosorbent assay（ELISA）. ResultCompared with the model group， the fecal water content and fecal number of mosapride group， M. officinalis cortex group and sweated M. officinalis cortex group were significantly increased（P<0.05）. At the phylum level， the top four species of flora abundance were Firmicutes， Bacteroidetes， Spirochaetes and Proteobacteria. Compared with the blank group， the proportion of Firmicutes in the model group was significantly reduced（P<0.05）， while the proportion of Spirochaetes was significantly increased（P<0.05）. Compared with the model group， the proportion of Firmicutes and Spirochaetes in M. officinalis cortex group and sweated M. officinalis cortex group tended to be similar to that in the blank group， and the proportion of Spirochaetes in sweated M. officinalis cortex group was lower than that of M. officinalis cortex group. At the family level， the top four species of flora abundance were Lactobacillaceae， S24_7， Ruminococcaceae， Bacteroidaceae， compared with the blank group， the proportion of Lactobacillaceae in the model group decreased significantly（P<0.05）， and its proportion in the M. officinalis cortex group and sweated M. officinalis cortex group increased significantly after administration（P<0.05）， and the flora structure of the two groups tended to be similar to that of the blank group. At the genus level， the top four species of flora abundance were Lactobacillus， Unspecified_S24_7， Bacteroides and Treponema. Compared with the blank group， the proportion of Lactobacillus in the model group decreased significantly（P<0.05）， while the proportion of Treponema increased significantly（P<0.05）. Compared with the model group， ratio of bacterial structure of Lactobacillus and Treponema in the M. officinalis cortex group and sweated M. officinalis cortex group tended to be similar to those in the blank group， indicating that M. officinalis cortex could restore the intestinal microbial structure of IBS-C rats before and after sweating. Compared with the model group， the 5-HT content in mosapride group was significantly reduced（P<0.05）， the contents of 5-HT and SP in the M. officinalis cortex group and sweated M. officinalis cortex group were significantly increased（P<0.01）， and the sweated M. officinalis cortex group was higher than the M. officinalis cortex group. ConclusionM. officinalis cortex can play a therapeutic role on IBS-C rats by regulating 5-HT pathway and intestinal flora structure before and after sweating.
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OBJECTIVE@#To observe the effects of moxibustion at "Baihui" (GV 20) and "Dazhui" (GV 14) at different time points on the serum level of β-endorphin (β-EP), substance P (SP) and expression of interleukin-1β (IL-1β) and cyclooxygenase-2 (COX-2) protein in brainstem in rats with migraine, and to explore the effect and mechanism of moxibustion in preventing and treating migraine.@*METHODS@#Forty male SD rats were randomly divided into a blank group, a model group, a prevention+treatment (PT) group and a treatment group, 10 rats in each group. Except the blank group, the rats in the remaining groups were injected with nitroglycerin subcutaneously to prepare migraine model. The rats in the PT group were treated with moxibustion 7 days before modeling (once a day) and 30 min after modeling, while the rats in the treatment group were treated with moxibustion 30 min after modeling. The "Baihui" (GV 20) and "Dazhui" (GV 14) were taken for 30 minutes each time. The behavioral scores in each group were observed before and after modeling. After intervention, ELISA method was used to detect the serum level of β-EP and SP; the immunohistochemistry method was used to detect the number of positive cells of IL-1β in brainstem; the Western blot method was used to detect the expression of COX-2 protein in brainstem.@*RESULTS@#Compared with the blank group, the behavioral scores in the model group were increased 0-30 min, 60-90 min and 90-120 min after modeling (P<0.01); compared with the model group, in the treatment group and the PT group, the behavioral scores were decreased 60-90 min and 90-120 min after modeling (P<0.01). Compared with the blank group, in the model group, the serum level of β-EP was decreased (P<0.01), while the serum level of SP, the number of positive cells of IL-1β in brainstem and the expression of COX-2 protein were increased (P<0.01). Compared with the model group, in the PT group and and the treatment group, the serum level of β-EP was increased (P<0.01), while the serum level of SP, the number of positive cells of IL-1β and the expression of COX-2 protein in brainstem were decreased (P<0.01, P<0.05). Compared with the treatment group, in the PT group, the serum level of β-EP was increased and COX-2 protein expression was decreased (P<0.05).@*CONCLUSION@#Moxibustion could effectively relieve migraine. The mechanism may be related to reduce the serum level of SP, IL-1β and COX-2 protein expression in brainstem, and increase the serum level of β-EP, and the optimal effect is observed in the PT group.
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Rats , Male , Animals , Moxibustion , Rats, Sprague-Dawley , Cyclooxygenase 2 , beta-Endorphin , Substance P , Interleukin-1beta , Migraine Disorders , Brain StemABSTRACT
Objective To investigate the expressions of substance P(SP)and neurokinin receptor(NK1R)in eosinophil-enriched cells from patients with atopic dermatitis(AD)so as to elucidate their possible roles in AD. Methods Blood samples were collected from healthy controls(HCs)and AD patients,and incubated with the extracts of Artemisia pollen,dust mite,and Platanus pollen for 1 h.The expressions of SP and NK1R in eosinophil-enriched cells were detected by flow cytometry.Results Level of NK1R in eosinophil-enriched cells from AD patients increased by 41% compared with that of HCs when cultured with the medium only(P= 0.001).In addition,the expression of SP in AD patients decreased by 1.17 folds(P＜0.001),and mean fluorescence intensity (MFI)of SP+eosinophils decreased by 55%(P＜0.001).However,allergens had little effect on the expressions of SP and NK1R in eosinophil-enriched cells from AD patients and HCs.Conclusion Upregulated expression of NK1R in AD indicates that eosinophil-derived NK1R may play an important role in AD.Antagonists or blockers of NK1R might be effective preparations for AD treatment.
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<p><b>OBJECTIVE</b>To explore the effect of transcutaneous electrical acupoint stimulation (TEAS) on postoperative analgesia of ureteroscopic holmium laser lithotripsy.</p><p><b>METHODS</b>One hundred and twenty adult patients, American Association of Anesthesiologists (ASA) Class Ⅰ or Ⅱ, scheduled to ureteroscopic holmium laser lithotripsy, were randomly assigned into an observation group and a control group, 60 cases in each one. The patients in the observation group were treated with TEAS for postoperative analgesia. TEAS was implemented at bilateral Shenshu (BL 23) and Yinlingquan (SP 9) at the time of back ward and postoperative 4 h, 8 h, 12 h. TEAS at 7:00, 11:00 and 15:00 at the above acupoints were used on the second and third days; while placebo (twice a day, 100 mg a time) was used. Tramadol hydrochloride tablets for postoperative analgesia were applied in the contnol group, twice a day, 100 mg a time, and electrode sheets without stimulation were put on Shenshu (BL 23) and Yinlingquan (SP 9). When analgesia was insufficient with the score of visual analogue scale (VAS)≥3, the patients were treated with tramadol tablets for remedy analgesia. The VAS score, the concentrations of serotonin (5-HT) and substance P (SP) in 3 mL venous blood at the time of back ward (T), postoperative 4 h (T), 12 h (T), 24 h (T), and 48 h (T) were detected respectively. The total amount of medication for remedy analgesia and the incidence of adverse reactions, such as nausea and vomiting within postoperative 48 h were compared between the two groups.</p><p><b>RESULTS</b>The VAS scores at T through T were lower than those at T in the two groups (all <0.05). Compared with the control group, the VAS scores at T through T in the observation group were lower (all <0.05). The total dose of remedy analgesic medicine within 48 h after operation in the observation group was lower than that in the control group (<0.05). Compared with the control group, the concentrations of 5-HT at T, T, T and SP at T through T were lower (all <0.05). The numbers of constipation, nausea and vomiting in the observation group were less than those in the control group (both <0.05).</p><p><b>CONCLUSION</b>TEAS can relieve the pain and reduce the total amount of analgesic medicine, the levels of substance causing pain and the incidence of adverse reactions after ureteroscopic holmium laser lithotripsy.</p>
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<p><b>OBJECTIVE</b>To dynamically observe the effects of electroacupuncture (EA) on repair of gastric mucosal lesion in rats with gastric ulcer, and to explore the time-effect relationship and molecular mechanism of EA for gastric ulcer.</p><p><b>METHODS</b>A total of 72 SD rats were randomly assigned to a normal group, a model group, a acupoint group and a sham acupoint group, and each group were further divided into a 1-day subgroup, a 4-day subgroup and a 7-day subgroup, 6 rats in each subgroup. The rat model of gastric ulcer was established by using intragastric administration of ethyl alcohol. The rats in the acupoint group were treated with EA at"Zusanli"(ST 36) and"Liangmen"(ST 21); the rats in the sham acupoint group were treated with EA at points 5 mm next to"Zusanli"(ST 36) and"Liangmen"(ST 21); the EA was given 30 min per treatment, once a day. The rats in the normal group and model group were treated with immobilization for 30 min per day, and no EA was given. PR-PCR method was applied to test the expression of proliferating cell nuclear antigen (PCNA) and substance P (SP); Western blot method was applied to test the expression of neurotensin (NT).</p><p><b>RESULTS</b>After 1-day treatment, the ulcer index in the model group was higher than that in the normal group (<0.01), and the expression of PCNA, SP and NT was decreased (<0.01, <0.05); compared with the model group and sham acupoint group, the ulcer index was decreased in the acupoint group (both <0.05), and the expression of PCNA and SP was up-regulated (all <0.05) while that of NT was up-regulated (both <0.01). After 4-day treatment, the ulcer index in the model group was reduced but still higher than that in the normal group (<0.05), and the expression of PCNA, SP and NT was up-regulated and higher than that in the normal group (all <0.01); the ulcer index in the acupoint group was similar to that in the normal group (>0.05), and the expression of PCNA and SP was lower than that in the model group (<0.01, <0.05), and the expression of NT was not significantly different from that in the model group (>0.05). After 7-day treatment, the differences of indexes above were not significant among the four groups (all >0.05).</p><p><b>CONCLUSION</b>EA at acupoints of stomach meridian has two-way regulation on PCNA and SP and improve the expression of NT in different pathological state of gastric ulcer, which could further improve the repair of gastric ulcer.</p>
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We investigated the secretion of Interleukin-8 (IL-8) from ginviva and periodontal ligament stimulated with Substance P (SP) and Calcitonin Gene-related Peptide (CGRP). Gingiva (GF), periodontal ligament (PDLF) and pulp (PF) tissues were collected from extracted intact 3rd molars. Cultured cells were stimulated with different concentrations of SP for 4 hrs, and stimulated with SP, CGRP and Tumor Necrosis Factor-alpha (TNF-alpha) for 8 hrs. Then RNase Protection Assay was carried out. ELISA was performed using supernatants of stimulated cells for quantitative analysis of IL-8. Results were assessed using student t-test with significance of P < 0.05. According to this study, the results were as follows: 1. IL-8 mRNA was detected in all type of cells studied (PF, GF and PDLF). 2. IL-8 mRNA expression was not increased after stimulating 4 hrs with SP (10(-5)M) and SP (10(-8)M) compared with Mock stimulation in all type of cells studied. 3. IL-8 mRNA expression was not increased after stimulating 8 hrs with SP (10(-4)M) and CGRP (10(-6)M) compared with Mock stimulation in all type of cells studied. 4. TNF-alpha(2 ng/ml) increased the expression of IL-8 mRNA in all kind of cells studied. 5. The secretion of IL-8 from GF was increased 8 hrs after the stimulation with CGRP (10(-6)M) (p < 0.05). 6. The secretion of IL-8 from PDLF was increased 8 hrs after the stimulation with SP (10(-4)M) (p < 0.05). Calcitonin Gene-related Peptide (CGRP) increased Interleukin-8 (IL-8) which plays an important role in chemotaxis of neutrophil in Calcitonin Gene-related Peptide (CGRP) gingival tissue, whereas Substance P increased the secretion of IL-8 from periodontal ligament.
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Humans , Calcitonin Gene-Related Peptide , Cells, Cultured , Chemotaxis , Enzyme-Linked Immunosorbent Assay , Gingiva , Interleukin-8 , Molar , Neuropeptides , Neutrophils , Periodontal Ligament , Ribonucleases , RNA, Messenger , Substance P , Tumor Necrosis Factor-alphaABSTRACT
@#Objective To approach the neurobiochemical mechanism of chronic central pain (CCP) after spinal cord injury (SCI). Methods 28 SD rats were divided into four groups, the normal group (group A), the pseudosurgery group (group B), and groups with CCP (group C) and without CCP (group D) after L1 spinal cord section injured with WADE method. T13 and L2 segments of rats' spinal cord were took and concentration changes of substance P (SP) in the spinal dorsal horn between two sections were examined by immunofluorescence histochemistry staining combined with confocal laser scanning microscope. Results Concentration of SP in the group D was decreased significantly compared with groups C,A and B (P<0.05-0.01), that of the group C was less than that of group A and B (P<0.05). Conclusion The rat model established by WADE method is proper to study CCP after SCI. SP in dorsal horn of spinal cord may inhibit the CCP after SCI in some degrees.
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Objective The aim of this study was to evaluate and to characterize the visceral hypersensitivity in non-erosive reflux disease(NERD)through the detection of esophageal distension-cerebral evoked potentials(ED-CEP)and calcitonin gene related peptide(CGRP)and substance P(SP)levels in esophageal mucosa.Methods NERD patients(N 26)and 12 controls were enrolled in this study from Oct.2004 to Mar.2005.Mechanical distention stimulation was performed using the balloon-affixed and polyvinyl multilumen catheter.The ED- CEP was recorded with a muti-channel international 10~20 system of electroencephalograph.The esophageal specimens were immune-histologically evaluated for CGRP/SP-stain positive contents.Results Esophageal distention may evoke recognizable and reproducible muti-peak CEP.CEP morphology of NERD patients was characterized by randomly distributed patterns,and the peak latencies for N1,P1,and N2 were significantly shorter compared with control group(P
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@#In order to observe the changes of myotension in the paralyzed limbs after subcutaneous injection(S.C)of capsaicin,we observed the changes of the spontaneous Electromyography(EMG)recorded in the quadriceps femoris before and after T-5 level spinal transection and found that the amplitude of EMG kept a low level in normal rats but increased progressively after operation.After treating the animals with capsaicin 200mg/kg(S.C)in three consecutive days,we found a much lower spontaneous EMG and a much higher pain threshold than that of the controls.Radioimmunnoassay detected that the content of SP in spinal dorsal horn was reduced by 38.4%,the general condition did not severely changed during the experiment.The results indicate that SP in the spinal dorsal horn participates in the regulation of myotension,and it is suggested that subcutancous injection of capsaicin may be one effective treatment to reduce the hypermyotension in the paralyzed limbs.
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Objective To study the effect and the mechanism of Tongxieyaofang (TXYF) on rat model of irritable bowel syndrome (IBS). Methods Model rats of IBS which were prepared by chronic stimulation in colon were randomly divided into groups: the normal control group (Group A), the model control group (Group B), the positive control group (Group C), the low dosage of TXYF group (Group D), the high dosage of TXYF group (Group E). Different treatments were given to every groups by ig administration for one month. The capability limens of the sacculus that caused abdomen-uplifting and back-arching and the times of contract of abdomen muscle in rats under different dilatations were observed. The levels of 5-hydroxytryptamine (5-HT), substance P (SP), and calcitonin gene related peptide (CGRP) were evaluated. Results Compared with the Group B, the TXYF could effectively improve the index of the behavior and electrophysiology, it also could lower the levels of 5-HT and SP, and increase the levels of CGRP of rat model of IBS with certain dose-effect relationship. Conclusion TXYF could lower the levels of 5-HT in serum and SP in plasma, and increase CGRP level of model rats of IBS, especially in high dosage. The mechanism of TXYF may be increasing the pain threshold of bowel, eliminating hypersusceptibility of bowel, and decreasing excitability of nerve cells by decreasing the 5-HT and SP levels in IBS rats.
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Objective To observe the effects of tetramethylpyrazine (TMP) on acute nociception in rat hindpaw induced by purine 2X (P2X) receptor agonists, such as adenosine triphosphate (ATP) and ?, ?-meATP, prostaglandin E 2 (PGE 2), and substance P (SP). Methods The effects of TMP administered intraplantarlly on the acute nociception induced by P2X receptor agonists, PGE 2, or SP in the rat hindpaw were investigated by the method of the behavioral study. Results TMP (10 mmol/L) significantly depressed the acute nociception induced by ATP (1 ?mol/L) or ?, ?-meATP (0.6 ?mol/L) in the rat hindpaw. TMP (10 mmol/L) could inhibit the acute nociception induced by PGE 2 (5 ?mol/L) or ?, ?-meATP (0.2 ?mol/L) coinjected with PGE 2 (5 ?mol/L). TMP (10 mmol/L) could not affect the acute nociception induced by ?, ?-meATP (0.2 ?mol/L) coinjected with SP (10 ?mol/L). TMP could not obviously affect the inflammatory edema in rat hindpaw induced by the local administration of PGE 2, SP, or ?, ?-meATP coinjected with PGE 2 or SP individually. Conclusion The antinociceptive effects of TMP may mainly be associated with inhibiting the transmission of nociceptive information mediated by P2X receptor activation.
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Objective To study the role of calcitonin-gene-related peptide (CGRP) and substance P (SP) in the pathogenesis of prurigo nodularis. Methods SABC immunohistochemical technique was used to examine the expression of CGRP and SP in lesional skin and uninvolved skin of 30 cases of prurigo nodularis and in skin of 15 normal controls. Results The strong expression of CGRP and SP was observed in the whole epidermis of lesional skin in 30 cases, with the expression surrounding cell membranes. The weak expression of CGRP and SP was observed in nonlesional and normal epidermis. CGRP and SP expression levels had significant differences in the lesional skin and in normal and uninvolved skin (P
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This experiment was carried out to demonstrate substance P (SP) and VIP in mast cells in human skin by double immunoenzymatic staining, and to examine SP, VIP and serotonin in mast cells in mouse skin by dual immunoflourescence method. The results were as follows:1. There were three types of mast cells in human skin by double immunoenzymatic staining: i. e. SP immunoreactive mast cells, VIP immunoreactive mast cells and mast cells with coexistence of SP and VIP. In comparison with toluidine blue staining, 8.32％ of mast cells were VIP immunoreactive; 14.51％ were SP immnuoreactive; 7.71％ were SP and VIP coexistence cells.2. Similar to human skin, SP, VIP immunoreactive mast cells and mast cells with coexistence of SP and VIP were found in mouse skin by dual immunoflourescence. They were 12.7％, 7.9％, and 10.3% of she toluidine bleu positive mast cells, respectively, which acounted to about 24.6％ of the toluidine blue positive mast cells.The possible significance of these results were discussed.