Substance P Content Research Articles

Tear film substance P: A potential biomarker for diabetic peripheral neuropathy

ObjectiveTo explore the changes that occur in the concentrations of substance P (SP) and calcitonin gene-related peptide (CGRP) in tears as a result of corneal denervation and its association with diabetic peripheral neuropathy (DPN). MethodsSixty-three individuals with type 1 diabetes/type 2 diabetes (T1D/T2D) and 34 age-matched healthy controls underwent a detailed assessment of neuropathy using the Total Neuropathy Score (TNS). The concentration of SP and CGRP in tears was measured by enzyme-linked immunosorbent assay. The corneal sub-basal nerve plexus was imaged using corneal confocal microscopy. Corneal nerve fibre length, fibre density, branch density, total branch density, nerve fractal dimension and inferior whorl length were quantified. ResultsIn T1D, the median [IQR] concentration of SP in tears was significantly reduced in those with DPN, (130 [61–692]pg/mL) compared to both control subjects (763 [405–1555]pg/mL, P < 0.01) and in those without DPN (914 [339–1832]pg/mL, P = 0.01); the concentration of CGRP was not changed. In T2D, there was no difference in neuropeptides between participants with diabetes and controls, regardless of neuropathic status. In T1D and T2D, corneal nerve parameters were significantly different between those with DPN or without DPN and controls. A significant correlation was noted between the concentration of tear film SP and TNS in T1D (r = −0.49; P < 0.001) and corneal nerve fibre density (r = 0.45; P < 0.001). The concentration of tear film CGRP was correlated significantly with the reduction of corneal nerve fibre density (r = 0.41; P = 0.01) in T1D. ConclusionTear film SP may provide a potential non-invasive biomarker for assessing neuropathy in T1D.

Substance P Saliva Reduction Predicts Pharyngeal Dysphagia in Parkinson's Disease.

Introduction: Although patients with Parkinson's disease (PD) often suffer from oropharyngeal dysphagia, knowledge about the underlying pathophysiological mechanisms is limited. Substance P (SP) is a localization-independent neurotransmitter of the entire nervous system. Reduced levels of SP were found in saliva of patients with impaired cough reflex and in advanced stages of PD. The aim of the study was to investigate SP in PD patients in order to gain further insights into the underlying pathophysiology of PD-related dysphagia and to evaluate the potential of SP as a biomarker for early dysphagia.Methods: Flexible endoscopic evaluation of swallowing (FEES) was used to objectively assess pharyngeal swallowing function. From a cohort of 105 consecutive PD patients 20 subjects were recruited: in 10 of them pharyngeal dysphagia was excluded by FEES, the other 10 subjects showed signs of early pharyngeal dysphagia defined as hypopharyngeal sensory deficit with mild to moderate vallecular residues after swallowing solid consistencies. Analysis of the Substance P level in saliva of the 20 included PD patients was performed in the clinical on state condition by ELISA-type immunoassay. Significant differences were calculated by using the Mann-Whitney test.Results: Twenty PD patients with a mean age of 69.5 ± 12.5 years (8 female) were included in the study. No significant differences were found regarding gender, age, UPDRS III, Hoehn and Yahr stage, disease duration, and Levodopa equivalent dose between the non-dysphagic and dysphagic subjects. Dysphagia was mainly characterized by unrecognized residues in the valleculae without any aspiration risk for all of the tested consistencies in FEES and was thereby scored as mild in all cases. Saliva SP concentrations were significantly lower in PD patients with pharyngeal dysphagia compared to those with a normal pharyngeal swallowing function (9,644 vs. 17,591 pg/mL; p = 0.001).Conclusion: Reduced saliva SP concentrations may predict early pharyngeal swallowing dysfunction in PD patients. This finding supports the hypothesis that an impaired SP mediated neurotransmission has a significant impact for the development of dysphagia in PD patients. Larger studies are needed to confirm SP as a clinical useful biomarker for early detection of PD-related dysphagia.

Neuroprotective effect of surround needling combined with acupoint injection on acute herpetic neuralgia

To explore the effect and mechanism of surround needling combined with acupoint injection on acute herpetic neuralgia (AHN). Ninety-nine patients with T6-T10 segment AHN were randomly divided into 3 groups, 33 cases in each group, including 2 cases dropped off in the surround needling group, 4 cases dropped off in the acupoint injection group, and 3 cases dropped off in the combined group. Oral valacyclovir was given in each group, 0.3 g each time, 2 times a day for 10 days. Oblique insertion of needle used at ashi points around the herpes in the surround needling group, and continuous wave was stimulated to tolerance for 20 min; the same acupoints were selected as the surround needling group, stimulated with the mixture injection of mecobalamin and lidocaine in the acupoint injection group; After the surround needling, acupoint injection was performed in the combined group. The treatment was given once a day, 14 times for a course, and one course was needed in all groups. The skin healing conditions (blistering, crusting, and dislocation time) of each group were compared after treatment. The pain scores, pain area and quality of life scores in each group were observed before and after treatment. The levels of neuron specific enolase (NSE), substance P (SP) and calcitonin gene-related peptide (CGRP) in the local blister fluid were measured before and after treatment in all groups. The blistering, crusting and dislocation time in the combined group were earlier than the other two groups (all P<0.05). The pain score and pain area in the each group were significantly lower than those before treatment, and the quality of life score was significantly higher than that before treatment (all P<0.05). The improvements of pain score and quality of life score in the combined group were more obvious than the other two groups (all P<0.05). After treatment, the levels of NSE, SP and CGRP in the local blister fluid in each groups were significantly lower than those before treatment (all P<0.05). The indexes in the combined group were significantly lower than those in the other two groups (all P<0.05). Both surround needling and acupoint injection have an adjuvant effect on AHN. The combination of the two is better, the skin is healed quickly, the analgesia is significant, and the contents of local NSE, SP and CGRP are significantly decreased. The mechanism of action is to exert neuroprotective effects.

Serum Levels of Neuropeptides in Cows with Left Abomasal Displacement.

Abomasal displacement (AD) to the left is a common disease in high-yielding dairy cows after parturition. In view of the previously reported changes in tissue neuropeptide concentrations in cows with AD, the primary aim of this study was to evaluate the effect of AD and breed on serum neuropeptide concentrations. For this purpose, blood samples of 33 German Holstein (GH) cows with AD, 36 healthy controls (GH), and 32 healthy German Fleckvieh (GF) cows were collected, and concentrations of substance P (SP), vasoactive intestinal polypeptide (VIP), and interleukin1β (IL-1β) were measured via commercially available ELISA kits. To examine the effect of AD, we compared GH cows with and without AD and observed no significant effects of AD on SP, VIP, or Il-1 β concentrations. To evaluate the effect of breed, we compared healthy GH with healthy GF cows and detected markedly higher VIP serum levels in the healthy GF cows (p < 0.01). No significant differences in SP or IL-1β were detected. According to our results, there seems to be no effect of AD on the serum concentrations of SP, VIP, or IL-1 β. In contrast, there seems to be a breed difference concerning serum VIP concentrations.

The potential of substance P to initiate and perpetuate cortical spreading depression (CSD) in rat in vivo

The tachykinin substance P (SP) increases neuronal excitability, participates in homeostatic control, but induces brain oedema after stroke or trauma. We asked whether SP is able to induce cortical spreading depression (CSD) which often aggravates stroke-induced pathology. In anesthetized rats we applied SP (10−5, 10−6, 10−7, or 10−8 mol/L) to a restricted cortical area and recorded CSDs there and in remote non-treated areas using microelectrodes. SP was either applied in artificial cerebrospinal fluid (ACSF), or in aqua to perform a preconditioning. Plasma extravasation in cortical grey matter was assessed with Evans Blue. Only SP dissolved in aqua induced self-regenerating CSDs. SP dissolved in ACSF did not ignite CSDs even when excitability was increased by acetate-preconditioning. Aqua alone elicited as few CSDs as the lowest concentration of SP. Local pretreatment with 250 nmol/L of a neurokinin 1 receptor antagonist prevented the SP-induced plasma extravasation, the initiation of CSDs by 10−5 mol/L SP diluted in aqua, and the initiation of CSDs by aqua alone, but did not suppress KCl-induced CSD. Thus neurokinin 1 receptor antagonists may be used to explore the involvement of SP in CSDs in clinical studies.

TRPA1 and substance P mediate stress induced duodenal lesions in water immersion restraint stress rat model.

Transient receptor potential ankyrin 1 (TRPA1) and substance P (SP), both expression in sensory neurons, have important roles in stress-induced duodenal lesions. The possible contribution of TRPA1 and SP to stress-induced duodenal lesions was explored by using the water immersion restraint stress (WIRS) rat model. Western blotting, Real-time polymerase chain reaction (RT-PCR), and immunohistochemistry assay were used to evaluate the changes of TRPA1and SP expression in the dorsal root ganglia (DRG, T8-11), the corresponding segment of the spinal cord (T8-11), and the duodenum in a duodenal lesions rat model. The SP concentrations of duodenal mucosa were investigated using an enzyme-linked immunosorbent assay (ELISA). Duodenal lesions were assessed according to histopathological changes. TRPA1 specific antagonist HC-030031 was intrathecally or intraperitoneally performed to suppress the expression of both TRPA1 and SP for evaluating the roles of TRPA1 and SP in duodenal lesions. In contrast to the control group, TRPA1 and substance P in the DRG (T8-11) and duodenum were up-regulated, and concentrations of SP in the duodenal mucosa were increased after WIRS (p<0.05), which are closely associated with duodenal lesions. SP concentrations in the duodenal mucosa were decreased and duodenal lesions were alleviated by pretreatment with TRPA1 antagonist HC-030031. We identified a protective role for HC-030031 in WIRS-induced duodenal lesions. Furthermore, we demonstrated that WIRS increased the concentrations of SP in the duodenal mucosa in a TRPA1-dependent manner. However, WIRS caused no significant changes of TRPA1 and SP in the spinal cord (T8-11) compared with the control group (p>0.05). Our study indicates that TRPA1 antagonist HC-030031 alleviates duodenal lesions. TRPA1 is activated and sensitized, therefore concomitant neuropeptide SP is released, which exerts a critical role in inducing and maintaining duodenal lesions following WIRS in rats. This provides evidence that neuroimmune interactions may control duodenal injury. TRPA1 may be a potential drug target to inhibit the development of duodenal lesions by stress-induced in patients.

Medial Habenula-Interpeduncular Nucleus Circuit Contributes to Anhedonia-Like Behavior in a Rat Model of Depression.

The habenula is a nuclear complex composed of the lateral habenula (LHb) and medial habenula (MHb), two distinct structures. Much progress has been made to emphasize the role of the LHb in the pathogenesis of depression. In contrast, relatively less research has focused on the MHb. However, in recent years, the role of the MHb has begun to gain increasing attention. The MHb connects to the interpeduncular nucleus (IPN) both morphologically and functionally. The MHb-IPN pathway plays an important role in regulating higher brain functions, including cognition, reward, and decision making. It indicates a role of the MHb in the pathogenesis of depression. Thus, we investigated the role of the MHb-IPN pathway in depression. MHb metabolic activity was increased in the chronic unpredictable mild stress (CUMS)-exposed rat model of depression. MHb lesions in the CUMS-exposed rats reversed anhedonia-like behavior, as observed in the sucrose preference test, and significantly downregulated the elevated metabolic activity of the IPN. Substance P (SP)-containing neurons of the MHb were found to innervate the IPN and to be the main source of SP in the IPN. SP content of IPN tissue of the CUMS-exposed rats was increased and MHb lesions reversed this change. In the in vitro experiment, firing rate recordings showed that SP perfusion increased the activity of IPN neurons. Our results suggest that hyperactivity of the MHb-IPN circuit is involved in the anhedonia-like behavior of depression, and that SP mediates the effect of the MHb on IPN neurons.

Evaluation of serum level of substance P and tissue distribution of NK-1 receptor in endometrial cancer.

Substance P (SP), a neuropeptide belonging to the tachykinin family, exerts different biological activities mainly through neurokinin-1 receptor (NK1R). The role of SP/NK1R system in tumoral growth and spread is reported in several cancers. We aimed to evaluate the serum SP concentration and NK1R tissue distribution in endometrial cancer, and to study the relationship between these factors with tumor size, lymph node involvement, disease stage and cancer grade. Recruiting 22 patients with endometrial cancer and 21 patients with leiomyoma as the control group, serum SP concentration was measured using an ELISA method, and NK1R tissue distributions were immunohistochemically analyzed. Serum SP concentration in patients was significantly higher than the control group (p-value = 0.005). The expression level of NK1R in tumoral tissue was more than normal tissue (p-value < 0.001). The NK1R expression had a significant relationship with lymph node involvement (p-value = 0.005) and disease stage (p-value = 0.017). The NK1R expression was higher in more advanced and less-differentiated tumors. SP/NK1R system seems to play a role in tumor growth and development in endometrial cancer. As well, the NK1R expression increased in endometrial cancer, and may be considered as a prognostic factor; but further studies are needed in this field.

Serum neuropeptide concentrations in cows with intrapartum uterine torsion

Intrapartum uterine torsion is a common cause of dystocia in dairy cows. To clarify the pathophysiology of this disease, the aim of the present study was to examine serum neuropeptide and cytokine concentrations of cows with intrapartum uterine torsion. Blood samples of 20 cows with uterine torsion, 36 healthy controls, and 15 intrapartum cows without uterine torsion were obtained. Concentrations for substance P (SP), vasoactive intestinal polypeptide (VIP), and interleukin-1β (IL-1β) were quantified by using commercially available ELISA kits. Significant differences between groups were observed for SP in cows with uterine torsion and cows calving normally (P < 0.01). Markedly greater SP concentrations were observed in calving cows than in cows with uterine torsion. Compared with healthy controls, there were greater SP concentrations during parturition (P < 0.01). No significant group differences were detected for VIP concentrations. Global differences were observed in IL-1β (P = 0.04). Large amounts of SP are released into the blood during parturition. Because SP is mainly present within the cervix and functions as a biomarker and mediator of pain, cows with uterine torsion are presumed to not experience as much pain as cows that are calving normally. Consistent with this, in cows with uterine torsion, there is a disturbance in the opening of the cervix, along with only mild signs of colic. Furthermore, significantly elevated IL-1β concentrations correlate with the inflammation that occurs in cases of uterine torsion. Further research is needed to support these findings and clarify the clinical relevance.

Role of neuropeptide substance P and the bone morphogenetic protein signaling pathway in osteogenic differentiation of ST2 cells

This study aimed to investigate the role and mechanism of neuropeptide substance P (SP) in ST2 cell (bone mesenchymal stem cells of mice) osteogenic differentiation to provide a basis for the treatment of temporomandibular joint osteoarthritis. Third-generation ST2 cells were cultured with different concentrations of SP (0, 10⁻¹⁰, 10⁻⁸, 10⁻⁶, and 10⁻⁵ mol·L⁻¹). After 24, 48, and 72 h, cell proliferation was detected by CCK-8. The ST2 cells were cultured with 10⁻⁶ mol·L⁻¹ SP for 1, 3, 5, and 7 days. Subsequently, the expression of alkaline phosphatase (ALP), collagen typeⅠ(CollaⅠ), and osteocalcin (OCN) in the culture supernatant was tested by enzyme-linked immunosorbent assay (ELISA). ALP activity was detected by immunofluorescence staining. The ST2 cells were cultured with SP, Noggin (inhibitor of the bone morphogenetic protein signaling pathway), SP+Noggin, and 2% fetal bovine serum, respectively. Finally, the expression of ALP, CollaⅠ, and OCN in the culture supernatant was tested by ELISA. CCK-8 showed that the effect of cell proliferation was most obvious when the SP concentration was 10⁻⁶ mol·L⁻¹ (P<0.01). The ELISA results demonstrated that ALP expression significantly increased at day 5 compared with that in the control group (P<0.01), whereas the expression of CollaⅠand OCN significantly increased at day 7 (P<0.05). Immunofluorescence results showed that ALP activity was strongest at day 5. The expression of ALP, CollaⅠ, and OCN decreased after Noggin addition (P<0.05). SP can promote the proliferation and osteogenic differentiation of ST2 cells, and the bone morphogenetic protein signaling pathway may be involved in this process.

Effects of castration and a lidocaine-plus-flunixin treatment on growth and indicators of pain, inflammation, and liver function in Korean cattle bull calves

The aim of this study was to determine the effects of castration and a lidocaine-plus-flunixin (LF) treatment on growth and indicators of pain, inflammation, and liver function in Korean cattle bull calves. Forty Korean cattle bull calves (body weight 197.0 ± 2.94 kg and age 6.3 ± 0.09 months) were each assigned to one of four treatments (n = 10 heads/group): no castration with no LF injection (NC-NLF); no castration with LF injection (NC-LF); castration with no LF injection (C-NLF); and castration with LF injection (C-LF). LF treatment included a local anesthetic lidocaine hydrochloride injection (12 mL of 2% in the scrotum) and intravenous injection of a non-steroidal anti-inflammatory drug, flunixin meglumine (2 mg/kg body weight of 50 mg/mL solution), immediately prior to castration. For the NLF groups, a 0.9% NaCl placebo solution was used. Castration was performed surgically using a Newberry knife and a Henderson castrating tool. Blood was collected immediately before castration and at h 0.5, h 6, d 1, d 3, d 7, and d 14 after castration. Feed intake was recorded daily, and body weight was measured on the day prior to the experiment and at d 14 after castration. Castration tended (P = 0.07) to decrease average daily weight gain, but LF treatment did not affect weight gain. Castration increased both circulating cortisol concentrations (P < 0.001) at h 0.5 after castration and substance P (SP) concentrations (P = 0.001) at h 6 after castration. However, the LF treatment did not significantly reduce cortisol and SP concentrations in castrated animals. Castration increased (P < 0.001) circulating haptoglobin (Hp) concentrations on d 1 and d 3 after castration, and LF treatment tended (P = 0.09) to decrease Hp concentrations on d 1 and decreased (P = 0.02) Hp concentrations on d 3. Castration did not affect glutamic oxaloacetic transaminase (GOT) and glutamic pyruvate transaminase (GPT) concentrations. LF treatment increased GOT concentrations at h 6 (P < 0.001), d 1 (P < 0.001), and d 3 (P = 0.003). LF treatment also increased GPT concentrations on d 1 (P = 0.006) and d 3 (P = 0.003). In conclusion, castration of bull calves resulted in time-sequential increases in circulating concentrations of cortisol at h 0.5, SP at h 6, and Hp on d 1. LF treatment did not significantly reduce elevated cortisol and SP, but tended to decrease elevated Hp concentrations in castrated animals. Our study demonstrates that LF treatment is not sufficient for the reduction of the indicators of pain and inflammation in castrated calves, suggesting that additional alleviation methods are required.

PB3. Pharyngeal Electrical Stimulation in neurointensive care patients suffering from severe post-stroke dysphagia – Post stimulation increase of salivary substance P level may indicate treatment success

Introduction Dysphagia is one of the most important and prognostically relevant complications of acute stroke. Pharyngeal Electrical Stimulation (PES) is a treatment device that enhances cortical reorganization for the restoration of swallowing function after cerebral injury. Furthermore, it was shown that PES leads to a temporary increase of Substance P (SP) level in saliva but not serum in healthy adults. The neuropeptide SP likely acts as a neurotransmitter in the pharyngeal mucosa and enhances the swallow and cough reflex. Post-stroke dysphagia may be related to reduced SP-levels. Here, we investigated the association of PES treatment in neurointensive care patients suffering from severe post-stroke dysphagia and salivary SP level, as well as a possible link between SP level and decannulation success after PES. Material and methods In this prospective single-center study, 23 tracheotomized stroke patients from our neurological intensive care unit were included. Due to severe and persisting dysphagia and according to the protocol for standardized endoscopic evaluation of tracheotomy decannulation in critically ill neurologic patients, participants could not be decannulated. Comorbidities that are known to likely cause dysphagia were not an exclusion criterion in contrast to other studies investigating on PES. 10 min of PES were applied on three consecutive days (= one treatment cycle). Three patients received multiple stimulation cycles. Saliva samples were collected prior to and directly after each treatment. To determine SP concentration, Enzyme-linked immunosorbent assay (ELISA) was performed. Results After the first treatment cycle 60% (n = 14) of all participants, respectively 73% (n = 7) of patients without comorbidities that likely cause dysphagia were decannulated. There was a close relation between increase of SP directly after the treatment and stimulation success (= decannulation). In 89% (n = 9) of patients without clinical improvement of dysphagia, SP levels remained stable or even decreased; 79% (n = 11) of successfully treated patients showed a post-stimulation increase of SP. Applying logistic regression analysis, increased SP level showed to be independently predictive of improved swallowing function after PES (p Conclusion The increase of salivary SP level following PES may indicate treatment success and SP therefore possibly acts as a biomarker of an improved swallowing function following PES.

Effect of Manual Acupuncture Preconditioning on Behavior and Contents of Serum CGRP, SP, IL-1 β and TNF-α Levels in Migraine Rats

To observe the effect of liver-soothing and mental-activity-regulating (LSMAR) needling on behavior reactions and contents of serum calcitonin gene-related peptide (CGRP), substance P (SP), interleukin-1 β (IL-1 β) and tumor necrosis factor-α (TNF-α) in migraine rats, so as to explore its mechanism underlying relief of migraine. Thirty-two male Wistar rats were randomly divided into control, model, LSMAR needling and conventional needling groups (n=8 rats in each). The migraine model was established by hypodermic injection of glyceryl trinitrate (5 mg/kg) at the napex. Acupuncture preconditioning was conducted once daily for 8 days before modeling. Acupuncture needles were inserted into "Baihui" (GV 20), and bilateral "Fengchi" (GB 20), "Neiguan" (PC 6) and "Taichong" (LR 3), manipulated for a while and retained for 30 min. The rats' behavioral changes (times of head scratching, tail-biting, cage-crawling, and to- and fro-movement in 30 min) were scored (one symptom = one point) before modeling, after modeling, and after intervention. The concentrations of CGRP, SP, IL-1 β, TNF-α in serum were assayed by enzyme-linked immunosorbent assay (ELISA). After modeling, the behavioral scores were significantly increased in the migraine model rats relevant to the control group (P<0.05). Whereas after the treatment, the behavior scores were considerably decreased during 60-90 min and 120-150 min of the measuring period in both LSMAR and conventional needling groups compared with the model group (P<0.05). The concentrations of CGRP, SP, IL-1 β and TNF-α were significantly higher in the model group than in the control group (P<0.05), and notably lower in both LSMAR and conventional needling groups than in the model group (P<0.05). The therapeutic effect of LSMAR was markedly superior to that of conventional needling in down-regulating the concentrations of CGRP, SP, IL-1 β and TNF-α (P<0.05). Acupuncture preconditioning can effectively relieve pain in migraine rats possibly by decreasing the concentrations of CGRP, SP, IL-1 β and TNF-α in the peripheral blood, and the therapeutic effect of LSMAR is evidently superior to that of conventional needling.

Truncation of neurokinin-1 receptor-Negative regulation of substance P signaling.

Substance P (SP) is a tachykinin peptide, which triggers intracellular signaling in the nervous and immune systems, as well as, other local and systemic events. The interaction between SP and its receptor, neurokinin-1 receptor (NK1R), results in major downstream cellular actions, which include changes in calcium fluxes, ERK, and p21-activated kinase phosphorylation and NFκB activation. Two naturally occurring variants of the NK1R, the full-length, 407 aa receptor (NK1R-F) and the truncated, 311 aa isoform (NK1R-T), mediate the actions of SP. Receptor truncation partially disrupts signaling motifs of the carboxyl tail, a critical site for mediating NK1R signaling, resulting in a "less-efficient" receptor. Although NK1R-F is the predominant isoform in the central and peripheral nervous systems, NK1R-T is expressed in several tissues and cells, which include monocytes, NK cells, and T-cells. The SP binding domain is not affected by truncation and this site is identical in both NK1R receptor isoforms. However, while cells expressing NK1R-F respond to nanomolar concentrations of SP, monocyte and macrophage activation, mediated through NK1R-T, requires micromolar concentrations of SP in order to elicit signaling responses. Elevated plasma levels of SP are associated with increased inflammatory responses and NK1R antagonists reduce inflammation and cytokine production in vivo. This mini review presents and discusses the novel hypothesis that the expression of NK1R-T on immune system cells prevents immune activation in a milieu, which usually contains low concentrations of SP and, thus, maintains immune homeostasis. In contrast, in the activated neuronal microenvironment, when SP levels reach the threshold at tissue sites, SP promotes immune activation and modulates monocyte/macrophage polarization.

Association of down-regulation of calcitonin gene-related peptide and substance P with increase of myocardial vulnerability in diabetic neuropathic rats

Diabetic patients present high co-morbidities of neuropathy and severer consequences of coronary heart disease. But the pathological mechanism is still unclear. Here we investigated a potential association of diabetic impairment of sensory nerves with increase of vulnerability of myocardium in acute myocardial ischemia/reperfusion. A rat model of diabetes mellitus was induced by high fat and sugar diet plus a small dose of streptozotocin. Impairment of sensory nerves was evaluated by measurement of changes in tail flick latency to noxious thermal stimulation and calcitonin gene-related peptide (CGRP) and substance P (SP) in the dorsal root ganglia (DRG) and the myocardium of the heart were examined. The myocardial injury was examined by infarct size, apoptosis ratio of cardiomyocytes and cardiac troponin I in the animals underwent acute myocardial ischemia (for 30min) and reperfusion (for 120min). The effects of CGRP and SP on cardiomyocyte injury induced by high glucose and hypoxia/reoxygenation were tested in cultured myocytes. The diabetic animals presented significant elevation of noxious thermal threshold with obvious reduction of the contents of CGRP and SP in the DRG and the myocardium. Importantly, the diabetic animals showed significant increases of infarct size, myocyte apoptosis and serum cardiac troponin I after acute myocardial ischemia/reperfusion, compared to the non-diabetic control. Furthermore, exogenously administered CGRP and SP attenuated the myocyte injury induced by the high concentration of glucose and hypoxia/reoxygenation. These findings suggested that impairment of sensory nerves with significant reduction of CGRP and SP in DRG, ventricular myocardium and serum may be associated with increase of myocardial vulnerability in acute myocardial ischemia/reperfusion in streptozotocin-induced diabetic rats.

Model for Studying Anti- Allergic Drugs for Allergic Conjunctivitis in Animals.

Allergic conjunctivitis (AC), which is characterized by ocular itching, hyperemia, and edema, deteriorates quality of life. In this study, effects of anti-allergic drugs were evaluated by assessing eye-scratching behavior, the number of eosinophils in conjunctiva epithelial tissues, and concentrations of chemical mediators in the tears of the guinea pig model of ovalbumin (OA)-induced AC.MethodologyOn day 0, 3-week-old guinea pigs were sensitized by OA subconjunctival injections. On days 15, 17, and 19, OA solution was administered. Anti-allergic eye drops were administered 5 and 15 min before the final OA challenge on day 19. Scratching behavior within 1 h after OA exposure was studied. Eosinophils in the conjunctiva were stained with Giemsa reagent. Histamine and substance P (SP) concentrations in tears were measured using ELISA.ResultsSubconjunctivally injected guinea pigs were observed for clinical symptoms. Scratching responses significantly reduced with ketotifen or olopatadine treatment. Eosinophil numbers reduced in animals treated with ketotifen, levocabastine, or tranilast. Histamine and/or SP concentrations in tears were inhibited by some of these anti-allergic drugs.ConclusionsIt is important to assess the anti-allergic AC drugs objectively because there are several of these drugs currently available. This model allows for an objective evaluation of anti-allergic drugs for AC.

Substance P promotes hepatic stellate cell proliferation and activation via the TGF-β1/Smad-3 signaling pathway

Prolonged activation and proliferation of hepatic stellate cells (HSCs) usually results in the initiation and progression of liver fibrosis following injury. Recent studies have shown that Substance P (SP) participates in the development of fibrosis. However, whether SP is involved in liver fibrosis, especially in the activation and proliferation of HSCs, is largely unknown. In the present study, we measured the effects of a series of concentrations of SP on the cell viability and activation of HSC-T6 cells and LX2 cells. The underlying mechanism was also investigated. We found that SP effectively increased cell viability, both in an MTT assay (p<0.05) and in a lactate dehydrogenase activity assay (LDH) (p<0.05). Moreover, SP upregulated the protein expression of α-SMA and Collagen I (both p<0.05) and decreased the release of lipid droplets (LDs) (p<0.05), all of which are associated with HSC activation. Apoptosis analysis revealed that SP can attenuate the increase of cell apoptosis induced by serum withdrawal (p<0.05). Furthermore, these effects were all blocked by an SP receptor antagonist, L732138. More importantly, L732138 decreased the activation of the TGF-β1/Smad3 signaling pathway, which is highly associated with liver fibrosis. Taken together, our results demonstrate that SP can promote HSC proliferation and induce HSC activation via the TGF-β1/Smad3 signaling pathway.

Nasal irrigation for the treatment of vasomotor rhinitis: a pilot study

Objective: To observe the therapeutic effect of simple 3.0% saline nasal irrigation and combined treatment of 3.0% saline nasal irrigation and budesonide nasal spray for vasomotor rhinitis (VMR), and explore the long-term effect for VMR. Through examination of levels of substance P (SP) and mucin (MUC)5B in nasal lavage fluid, the mechanisms of nasal irrigation treatment for VMR was discussed. Methods: One hundred and one patients from Department of Otorhinolaryngology Head and Neck Surgery, Huashan Hospital of Fudan University with VMR were randomly divided into 4 groups. The number of patients was 24 in control group, 25 in budesonide nasal spray treatment group (budesonide group), 25 in nasal irrigation treatment group (nasal irrigation group) and 27 in budesonide nasal spray + nasal irrigation group (combined treatment group). Control patients were left untreated. Budesonide group was under budesonide nasal spray treatment, nasal irrigation group was treated using 3.0% saline with a temperature of 40℃ and combined treatment group was given both treatments. The duration of the intervention period was 3 months (90 days). Visual Analog Scale (VAS) was used to evaluate nasal symptoms, and the health-related quality of life was assessed using the 12-item Short Form Health Survey version 2.0 (SF-12v2). Enzyme-linked immunosorbent assay (ELISA) was used to assess the contents of SP and MUC5B in nasal lavage fluid before and after 3-month treatments in budesonide and nasal irrigation group in the study. MUC5B in nasal lavage fluid after the SP challenge and anticholinergic drug intervention in control group were also evaluated with ELISA. Results: Nighty out of 101 patients completed the study. In the budesonide and combined treatment group after relevant interventions, the total VAS score of nasal symptoms decreased (5.91±0.21 vs 3.82±0.15, 6.18±0.17 vs 3.92±0.15, t value was 8.193, 10.060, respectively, all P<0.05) and SF-12v2 score increased (146.00±1.23 vs 152.30±0.97, 146.00±1.08 vs 155.40±0.90, t value was 3.982, 6.697, respectively, all P<0.05), with both scores showed no significant differences in the nasal irrigation group (5.96±0.17 vs 5.72±0.15, 146.10±1.17 vs 147.00±0.94, t value was 1.038, 0.607, respectively, all P>0.05) after the first month. In the budesonide and combined treatment group after relevant interventions, the total VAS score of nasal symptoms decreased (5.91±0.21 vs 5.05±0.15, 6.18±0.17 vs 5.10±0.12, t value was 3.374, 5.351, respectively, all P<0.05) and SF-12v2 score increased (146.00±1.23 vs 150.90±0.76, 146.00±1.08 vs 153.60±0.94, t value was 3.373, 5.343, respectively, all P<0.05), with both scores showed no significant differences in the nasal irrigation group (5.96±0.17 vs 5.78±0.17, 146.10±1.17 vs 148.10±0.80, t value was 0.716, 1.438, respectively, all P>0.05) after the second month. By the end of the third month, in nasal irrigation and combined treatment group, the VAS score was diminished (5.96±0.17 vs 4.80±0.12, 6.18±0.17 vs 4.44±0.13, t value was 5.485, 8.264, respectively, all P<0.05) and SF-12v2 score was elevated (146.10±1.17 vs 150.80±0.96, 146.00±1.08 vs 152.90±0.85, t value was 3.163, 5.008, respectively, all P<0.05), but there were no significant differences in budesonide group (5.91±0.21 vs 5.68±0.18, 146.00±1.23 vs 148.40±0.85, t value was 0.819, 1.587, respectively, all P>0.05). Additionally, SP in nasal lavage fluid decreased and MUC5B showed no statistical changes in budesonide group after three months, however, SP showed no any changes and MUC5B reduced significantly in nasal lavage fluid in nasal irrigation group. Furthermore, the anticholinergic drug could not decrease the concentration of MUC5B after the SP challenge in nasal cavity in control group. Conclusions: The therapeutic effect of simple nasal irrigation with 3.0% saline or combined treatment of 3.0% saline nasal irrigation and nasal corticosteroids is superior to simple nasal corticosteroids. Nasal corticosteroids plays a role in the inhibition of sensory nerve endings in nasal mucosa, but neurotransmitter plays a limited role in the pathogenesis of VMR.

Effects of different acupuncture-moxibustion methods on the plasma SP content and expression of colonic SP mRNA of rats with functional constipation

ObjectiveTo explore the effects of different acupuncture-moxibustion methods on the plasma substance P (SP) content, and the expression of colonic SP mRNA of the rats with functional constipation. MethodsSixty healthy male SD rats were divided into six groups according to the random number table, including blank control group (group B, n=8), model group (group M, n=11), drug control group (group D, n=8), filiform needle group (group F, n=11), electroacupuncture group (group E, n=11) and moxibustion group (group MB, n=11). The rats in group B and group M accepted no intervention, gavage with cisapride suspension was conducted in group D, and electroacupuncture, filiform needling and moxibustion therapies were carried out in group E, group F and group MB for 6 days. After treatment, the stool quantity and fecal moisture content during 24 h of the rats were observed in each group, colon tissue structure and the acidic mucus in mucosa layer were observed, plasma SP content was detected and the expression of colonic SP mRNA was detected. ResultsCompared with group B, the stool quantity and fecal moisture content during 24 h of the rats in group M decreased significantly (P<0.01); compared with group M, the stool quantity during 24 h of the rats in each group increased (P<0.05 or 0.01), and the fecal moisture content during 24 h of the rats in each group also increased significantly except group MB (P<0.01); compared with group D, the fecal moisture content during 24 h of the rats in group MB decreased (P<0.01); according to the comparison among the groups with different acupuncture-moxibustion methods, there was no difference in the stool quantity during 24 h of the rats in each group (P>0.05), the fecal moisture contents during 24 h of the rats in group F and group E were higher than that of group MB (P<0.01), and there was no difference between group F and group E (P>0.05). HE staining of colon tissue of rats showed that the structure was normal and complete, and there was no difference among each group. PAS staining showed that the acidic mucus level of the rats in group M decreased significantly when compared with group B; compared with group M, the acidic mucus level of the rats in group D, fgroup F, group E and group MB increased in varying degrees. Compared with group B, the plasma SP content of the rats in group M decreased (P<0.05); compared with group M, the plasma SP contents of the rats in group F and group MB increased (P<0.01); the plasma SP contents of the rats in group F, group MB and group E were higher than that of group D (P<0.05 or 0.01); according to the comparison among the groups with different acupuncture-moxibustion methods, the plasma SP content of the rats in group F was higher than that of group MB and electro-acupuncture group (P<0.05), and there was no difference between group MB and group E. Compared with group B, the expression level of colonic SP mRNA of the rats in group M elevated (P<0.05); compared with group M, the expression level of colonic SP mRNA of the rats in the four intervention groups declined (P<0.05); compared with drug group D, there was no difference among group F, group E and group MB (P>0.1); according to the comparison among the groups with different intervention methods, the expression level of colonic SP mRNA of the rats in group MB was lower than that of group F (P<0.05). ConclusionThe different efficacy of different acupuncture-moxibustion methods in treatment of functional constipation may be related with the mechanism of regulation of plasma SP content and the expression level of colonic SP mRNA.

Effects of transdermal flunixin meglumine on pain biomarkers at dehorning in calves.

The objective of this study was to evaluate the analgesic properties of transdermal flunixin meglumine when given at the time of dehorning on pain biomarkers. Twenty-four weaned male Holstein calves, 6 to 8 wk of age were enrolled into the study. The calves were randomly assigned to 1 of 3 treatment groups: 1) transdermal flunixin and dehorn (DH-FLU); 2) transdermal flunixin and sham dehorn (SHAM-FLU); and 3) placebo and dehorn (DH-PLBO). Transdermal flunixin at a label dose of 3.33 mg/kg (or placebo at an equivalent volume) was administered as a pour-on along the top-line of the calves in each treatment group concurrently with electrocautery dehorning or sham dehorning. Biomarker parameters collected and analyzed included: infrared thermography (IRT), mechanical nociception threshold (MNT), plasma cortisol, and substance P (SP). There were no differences in maximal temperatures detected for the IRT measurements of the medial canthus of the eye for the DH groups. Mean control point MNT measurements at 48 h were 3.14 kgF, 3.46 kgF, and 1.43 kgF for the DH-FLU, Sham-FLU, and DH-PLBO groups, respectively (P = 0.0001). No other differences of MNT were detected between the dehorned groups for the other test sites and time points. Plasma cortisol reached peak concentration at 20 min postdehorning for the DH-FLU and DH-PLBO groups and 10 min for SHAM-FLU group. Peak plasma cortisol concentrations were 32.0 ng/mL, 12.7 ng/mL, and 28.8 ng/mL for the DH-FLU, SHAM-FLU, and DH-PLBO groups, respectively. Cortisol concentrations were lower for the DH-FLU group at 90 min postdehorning compared to the SHAM-FLU and DH-PLBO groups ( = 0.04). Area under the effect curve (AUEC) were similar for all groups ( = 0.93). No statistical differences in SP concentrations between groups were detected for any of the time points. In conclusion, transdermal flunixin meglumine given at the time of dehorning did not provide substantial analgesia based on the pain biomarkers investigated. Further investigation into its role as part of a multimodal analgesic plan is warranted.