<p>OBJECTIVESTo investigate the hair growth-promoting effect of Miscanthus sinensis var. purpurascens (MSP) flower extracton on in vitro and in vivo models.p><p>METHODSMSP flower extract was extracted in 99.9% methanol and applied to examine the proliferation of human dermal papilla cells (hDPCs) in vitro at the dose of 3.92-62.50 μg/mL and hair growth of C57BL/6 mice in vivo at the dose of 1000 μg/mL. The expression of transforming growth factor β1 (TGF-β1), hepatocyte growth factor (HGF), β-catenin, substance P was measured by relative quantitative realtime polymerase chain reaction. Histopathological and immunohistochemical analysis were performed.p><p>RESULTSMSP (7.81 μg/mL) down-regulated TGF-β1 and up-regulated HGF and β-catenin in hDPCs (P<0.01). MSP (1000 μg/mL)-treated mice showed the earlier transition of hair follicles from the telogen to the anagen phase. The number of mast cells was lower in the MSP-treated mice than in other groups (P<0.05 vs. NCS group). Substance P and TGF-β1 were expressed in hair follicles and skin of the MSP group lower than that in negative control. Stem cell factor in hair follicles was up-regulated in the MSP-treated mice (P<0.01).p><p>CONCLUSIONSThe MSP flower extract may have hair growth-promotion activities.p>
Animals ; Antioxidants ; pharmacology ; Cell Count ; Cell Proliferation ; drug effects ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Female ; Flowers ; chemistry ; Hair Follicle ; cytology ; drug effects ; growth & development ; Hepatocyte Growth Factor ; metabolism ; Humans ; Mast Cells ; cytology ; Mice, Inbred C57BL ; Phosphorylation ; drug effects ; Plant Extracts ; pharmacology ; Poaceae ; chemistry ; RNA, Messenger ; genetics ; metabolism ; Skin ; metabolism ; Stem Cell Factor ; metabolism ; Stress, Psychological ; pathology ; Substance P ; metabolism ; Transforming Growth Factor beta ; genetics ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; beta Catenin ; metabolism

OBJECTIVE: The goal of this study was to study the mechanism of substance P (SP)-mediated the neural control of mast cell (MC) degranulation. METHOD: Bone marrow mast cells from mice were cultured with stem cell factor (SCF), IL-3 and IL-4 (group A) and SCF, IL-3 (group B) for four weeks. Then the cells were harvested and reserved for studies. Western Blot hybridization technique was used to detect the expression of FcεR I α and NK-1R on MCs from the two groups. Then such cells were activated with SP (0, 0. 01, 0. 10, 1. 00, 10. 00 µg/ml, respectively) for 30 min. The histamine released into the supernatant and stored in the protoplasm was quantified by enzyme linked immunosorbent assay (ELISA). And the percentage of histamine release was calculated as a percent of total histamine content. RESULT: The expressions of FcεR I α and NK-1R on these mast cells in group A were statistically higher than in group B (P<0. 05). The MCs from two groups can be actived when stimulated by SP, but the level of MC degranulation in group A was higher than group B (P<0. 05). CONCLUSION Neuropeptide may stimulate MC degranulation through immunological and non-immunological pathways. In summary, the current study provides us with better understanding of the mechanism of neuropeptide-controlled MC deranulation, and this should be helpful for the further research involved in the mechanism and treatmemt of airway hyper-reactivity.
Animals ; Bone Marrow Cells ; Cell Degranulation ; Cells, Cultured ; Culture Media ; chemistry ; Histamine ; metabolism ; Interleukin-3 ; pharmacology ; Interleukin-4 ; pharmacology ; Mast Cells ; cytology ; metabolism ; Mice ; Stem Cell Factor ; pharmacology ; Substance P ; pharmacology

<p>BACKGROUNDThe possible role of substance P (SP) during wound healing has been the primary research focus in recent years, but its effect on the healing process after bile duct injury is little understood. This study aimed to investigate the effects of SP on growth of fibroblast-like cells derived from rabbit bile duct.p><p>METHODSFibroblast-like cells derived from rabbit bile duct were identified and divided randomly into control and experimental groups. SP-treated cells at different concentrations of 10(-9)-10(-5) mol/L and control group were incubated, respectively, for 48 hours. After incubating, the effects of SP on cell proliferation were assessed by cell counts and MTT test. Apoptosis rate (AR) of cells was measured by flow cytometry.p><p>RESULTSCultured rabbit bile duct cells were fibroblast-like in morphology, and these cells were stained positively for vimentin and negatively for desmin. After SP was added to nonconfluent cells for 48 hours, cell numbers were significantly increased in experimental groups than in controls (P < 0.05). The maximum stimulation of cell proliferation was achieved at SP of 10(-5) mol/L. Bile duct fibroblast-like cells in the SP group showed a higher proliferating activity and lower AR than those in the control group or in the SP + Spantide group (P < 0.05). Spantide partly inhibited the effects of SP on fibroblast-like cells. Examination under transmission electron microscopy revealed rough endoplasmic reticulum and prominent Golgi complexes after SP treatment.p><p>CONCLUSIONSSP has a growth regulatory property on cultivated bile duct fibroblast-like cells in vitro, suggesting that SP may involve in wound healing after bile duct injury by promoting wound fibroblast proliferation and inhibiting apoptosis and participate in pathological scar formation.p>
Animals ; Bile Ducts ; cytology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Fibroblasts ; cytology ; drug effects ; Male ; Rabbits ; Substance P ; pharmacology

BACKGROUND/AIMS: DA-9701, a standardized extract of Pharbitis Semen and Corydalis Tuber, is a new prokinetic agent that exhibits an analgesic effect on the abdomen. We investigated whether DA-9701 affects visceral pain induced by colorectal distension (CRD) in rats. METHODS: A total of 21 rats were divided into three groups: group A (no CRD+no drug), group B (CRD+no drug), and group C (CRD+DA-9701). Expression of pain-related factors, substance P (SP), c-fos, and phosphorylated extracellular signal-regulated kinase (p-ERK) in the dorsal root ganglion (DRG) and spinal cord was determined by immunohistochemical staining and Western blotting. RESULTS: The proportions of neurons in the DRG and spinal cord expressing SP, c-fos, and p-ERK were higher in group B than in group A. In the group C, the proportion of neurons in the DRG and spinal cord expressing p-ERK was lower than that in group B. Western blot results for p-ERK in the spinal cord indicated a higher level of expression in group B than in group A and a lower level of expression in group C than in group B. CONCLUSIONS: DA-9701 may decrease visceral pain via the downregulation of p-ERK in the DRG and spinal cord.
Analgesics/*pharmacology ; Animals ; Colon ; Dilatation, Pathologic/physiopathology ; Down-Regulation ; Extracellular Signal-Regulated MAP Kinases/drug effects/*metabolism ; Ganglia, Spinal/drug effects/*metabolism ; Male ; Phytotherapy/methods ; Plant Preparations/*pharmacology ; Proto-Oncogene Proteins c-fos/metabolism ; Rats ; Rats, Sprague-Dawley ; Rectum ; Spinal Cord/drug effects/*metabolism ; Substance P/metabolism ; Visceral Pain/prevention & control

<p>OBJECTIVETo observe the analgesic effect of triptolide (TP) of high, middle and low doses on rats with adjuvant arthritis (AA), and the expressions of inducible nitric oxide synthase (iNOS) and substance P (SP) in spinal dorsal horn and dorsal root ganglion (DRG) of corresponding sections, in order to discuss the possible mechanism for the analgesic effect of TP on rats with adjuvant arthritis.p><p>METHODFifty SD rats were selected and randomly divided into the normal group (group A), the model group (group B), and TP low (group C), middle (group D), high (group E) dose groups. Except for the group A, all of the remaining groups were injected with 0.1 mL of Freund's complete adjuvant through their right rear toes to establish the model. At 14 d after the model establishment, rats in C, D and E groups were intraperitoneally injected with different doses of TP (0.1 mg x kg(-1) for the group C, 0.2 mg x kg(-1) for the group D, 0.4 mg x kg(-1) for the group E) once a day for 9 days. Then the 50% mechanical withdraw threshold (MWT) was determined. And the expressions of iNOS and SP in lumbar5 (L5) spinal dorsal horn and DRG were detected with the immunohistochemical method.p><p>RESULTThe 50% MWT of rats in the group B was significantly lower than that of the group A (P < 0.01). After being treated with TP, the Thermal withdrawal latencies of groups C, D and E were significantly higher than that of the group B (P < 0.01). TP could notably increase the MWT of AA rats, with a certain dose-effect relationship. The immunohistochemical results indicated that the iNOS and SP expressions significantly increased in the group B (P < 0.01), while the positive expression levels of iNOS and SP in groups C, D and E were significantly lower than that of the group B (P < 0.01), with a certain dose-effect relationship.p><p>CONCLUSIONTP shows a good analgesic effect on AA, and could inhibit the iNOS and SP expressions in spinal dorsal horn and DRG in rats with adjuvant arthritis, which may be one of action mechanisms for the analgesic effect of TP.p>
Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Arthritis, Experimental ; drug therapy ; metabolism ; physiopathology ; Diterpenes ; pharmacology ; Dose-Response Relationship, Drug ; Epoxy Compounds ; pharmacology ; Female ; Ganglia, Spinal ; drug effects ; metabolism ; Immunohistochemistry ; Male ; Nitric Oxide Synthase Type II ; biosynthesis ; Pain Measurement ; methods ; Phenanthrenes ; pharmacology ; Phytotherapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spinal Cord ; drug effects ; metabolism ; Substance P ; biosynthesis ; Time Factors ; Treatment Outcome ; Tripterygium ; chemistry

<p>OBJECTIVETo study the changes in plasma substance P (SP) and calcitonin gene-related peptide (CGRP) levels in children with acute asthma before and after gamma-aminobutyric acid (GABA) treatment.p><p>METHODSSeventy-five children with asthma were randomly assigned to GABA treatment (n=36) and control groups (n=39). Both groups were given conventional treatment for asthma. Besides the conventional treatment, the treatment group was administered with oral GABA (25-30 mg/kg•d). Plasma content of SP and CGRP was measured using ELISA before treatment and after remission.p><p>RESULTSThere were no significant differences in plasma content of SP and CGRP between the GABA treatment and control groups (P＞0.05) before treatment. Plasma content of SP and CGRP in the GABA treatment group was significantly lower than the control group (SP: 57±15 pg/mL vs 127±12 pg/mL; CGRP: 23±10 pg/mL vs 42±8 pg/mL) after remission (P<0.01). Plasma content of SP and CGRP after remission was significantly lower than before treatment (P<0.01) in both groups. There was a significantly positive correlation between plasma SP and CGRP content in asthmatic children (r=0.792, P<0.01).p><p>CONCLUSIONSGABA can significantly decrease plasma levels of SP and CGRP in children suffering from acute asthma.p>
Asthma ; blood ; drug therapy ; Calcitonin Gene-Related Peptide ; blood ; Child ; Child, Preschool ; Female ; Humans ; Male ; Substance P ; blood ; gamma-Aminobutyric Acid ; pharmacology ; therapeutic use

<p>OBJECTIVETo observe the effect of sensory neuropeptide substance P combined with epidermal stem cells (ESC) on wound healing and nerve regeneration in diabetic rats.p><p>METHODSESC that had been isolated from SD rats were identified and cultured in vitro, and they were inoculated onto nourishing layer of amniotic membrane to construct amniotic membrane-ESC. Four full-thickness skin wounds were produced on the back of each of 48 diabetic rats. The resulted 192 wounds were randomly divided into ESC + substance P group, ESC group, substance P group, and control group according to the lottery method, with 48 wounds in each group. Wounds in ESC + substance P group and ESC group were transplanted with amniotic membrane-ESC, and those in substance P group and control group were transplanted with amniotic membrane. After transplantation, 250 µL substance P in the concentration of 1 × 10(-7) mol/L was injected around and into the middle of the wounds in ESC + substance P group and substance P group, 2 times a day, and continued for 4 days, while 250 µL PBS solution was injected in the above-mentioned position in ESC group and control group as control, 2 times a day, and continued for 4 days. On post injury day (PID) 4, 7, 10, 14, 17, and 23, the wound healing rate (with 8 wounds at each time point) was observed and determined, and changes in wound tissue structure were observed with HE staining. On PID 4, 7, and 10, collagen distribution in wound tissue was observed with Masson staining, and type I and type III collagen deposition in wound tissue was respectively observed after immunohistochemical staining. The distribution of protein gene product 9.5 (PGP 9.5) and regeneration of substance P positive nerve fibers in wound tissue were observed with immunohistochemical staining on PID 14 and 23. Data were processed with one-way analysis of variance and t test.p><p>RESULTS(1) The wound healing rate in ESC + substance P group reached 100.0% on PID 14, which was obviously earlier than that in ESC group, substance P group, and control group, healing was respectively observed on PID 17, 17, and 23. The wound healing quality in ESC + substance P group was better than that in the other three groups as shown by HE staining. (2) On PID 10, collagen that was darkly stained and widely distributed was observed in wound tissue of ESC + substance P group and substance P group, while collagen in the other two groups was lightly stained and narrowly distributed. Deposition quantity of type I collagen gradually increased, and that of type III collagen gradually decreased in the wounds of each group over time. On PID 4, 7, and 10, distribution amount of type I collagen in wound tissue of ESC + substance P group was significantly higher than that in ESC group (with t value respectively 32.72, 118.21, 26.71, P values all below 0.01) and control group (with t value respectively 44.37, 22.76, 30.32, P values all below 0.01), while there was no significance between ESC + substance P group and substance P group. On PID 4, 7, and 10, distribution amount of type III collagen in wound tissue of ESC + substance P group was significantly higher than that in ESC group (with t value respectively 32.27, 28.68, 14.51, P values all below 0.01) and control group (with t value respectively 35.68, 22.52, 22.24, P values all below 0.01). (3) A large amount of PGP 9.5 and regeneration of substance P positive nerve fibers, and some peripheral nerve fibers in deep wound extending to epidermis were observed in wound tissue of ESC + substance P group and substance P group. A small amount of PGP 9.5 and regeneration of substance P positive nerve fibers without peripheral nerve fibers extending to epidermis were observed in deep wound tissue of ESC group and control group. On PID 14, 23, ratios of area of PGP 9.5 positive nerve fiber in the wounds of ESC + substance P group were (3.86 ± 0.25)% and (7.03 ± 0.28)%, and they were significantly higher than those of ESC group [(1.48 ± 0.30)%, (3.01 ± 0.43)%, with t value respectively 23.95, 30.27, P values all below 0.01] and control group [(1.46 ± 0.23)%, (2.84 ± 0.29)%, with t value respectively 27.35, 40.32, P values all below 0.01]. On PID 14, 23, ratios of substance P positive nerve fiber area in the wounds of ESC + substance P group were (2.01 ± 0.14)% and (1.19 ± 0.11)%, which were obviously higher than those of ESC group [(0.85 ± 0.17)%, (1.34 ± 0.21)%, with t value respectively 20.50, 2.60, P < 0.05 or P < 0.01] and control group [(0.74 ± 0.15)%, (1.30 ± 0.17)%, with t value respectively 23.98, 2.41, P < 0.05 or P < 0.01].p><p>CONCLUSIONSJoint application of substance P and ESC can effectively promote healing of wound and nerve regeneration in diabetic rats.p>
Animals ; Diabetes Mellitus, Experimental ; pathology ; Epithelial Cells ; cytology ; Nerve Regeneration ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; cytology ; Substance P ; pharmacology ; therapeutic use ; Wound Healing

This study was purposed to investigate the reversal effect of gambogic acid (GA) on multidrug resistance of K562/A02 cells and its mechanism. The IC(50) (half maximal inhibitory concentration) of adriamycin (ADM) was evaluated by MTT. Cell apoptosis was detected by flow cytometry. Morphological changes of K562/A02 cells were observed by fluorescent microscopy with DAPI staining. The expressions of Survivin and P-gp were determined by Western blot. The results showed that the IC(50) of ADM on K562 and K562/A02 cell proliferation were (1.42 ± 0.07) µg/ml and (28.42 ± 1.40) µg/ml respectively. GA ≤ 0.0625 µmol/L had no inhibitory effect on proliferation of K562 and K562/A02. 0.0625 µmol/L GA could enhance the sensitivity of K562/A02 cells to ADM (P < 0.05) and the reversal multiples was 1.53. The apoptotic rate was raised after treating with ADM combined with 0.0625 µmol/L GA for 48 h (P < 0.05). Morphological differences were typical and obvious between cells of control and treated groups under fluorescence microscopy using DAPI staining. After treating K562/A02 cells with ADM combined with 0.0625 µmol/L GA for 48 h, the expressions of Survivin and P-gp were down-regulated at protein levels. It is concluded that GA can enhance the sensitivity of K562/A02 cells to ADM, which may be related to increasing cell apoptosis and down-regulating expressions of Survivin and P-gp.
Apoptosis ; drug effects ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Gene Expression Regulation, Leukemic ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; K562 Cells ; Substance P ; metabolism ; Xanthones ; pharmacology

<p>OBJECTIVETo investigate the molecular pathway of substance P (SP) to induce osteoblastic differentiation.p><p>METHODSMesenchymal stem cells were isolated and cultured. The cultures were divided into four groups with Group A (control group) cultured without any factors, Group B cultured with SP, Group C cultured with SP and SP receptor neurokinin-1 (NK1) antagonist, and Group D cultured with SP NK1 antagonist respectively to induce osteoblastic cells differentiation. Osterix gene expression was detected by reverse transcription-polymerase chain reaction (RT-PCR) for three times after 1-2 weeks of cultivation and the results were analyzed by one-way analysis of variance (ANOVA).p><p>RESULTSThe log phase of bone marrow stromal cells appeared at 4-6 days. ALP staining revealed that the majority of cells, more than 95%, were positive and small blue-purple granules were found in the cytoplasm. And Group B, treated with SP, showed a higher level of ALP activity than the other three groups. Meanwhile, RT-PCR found that Osterix expression in Group B was obviously up-regulated, compared with other groups. But Osterix expression in Group D had no remarkable differences, compared with the controls.p><p>CONCLUSIONSSP can up-regulate Osterix gene expression to stimulate differentiation of mesenchymal stem cells into osteoblastic cells at the final stage. The regulatory effect of SP on Osterix expression was dependant on SP NK1 receptors.p>
Alkaline Phosphatase ; analysis ; Animals ; Cell Differentiation ; drug effects ; Gene Expression Regulation ; drug effects ; Osteoblasts ; cytology ; drug effects ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Substance P ; pharmacology ; Transcription Factors ; genetics ; Up-Regulation

<p>OBJECTIVETo investigate the effect of botulinum toxin type A on the expression of substance P (SP), calcitonin gene-related peptide (CGRP), transforming growth factor beta-1 (TGF-beta1) and alpha smooth muscle actin A (alpha-SMA) in wound healing.p><p>METHODS60 rats were randomly divided into group C (control) group L (low-dose) and group H (high-dose), with 20 rats in each group. The wound-healing model was established by excision of four full-thickness skin (1 cm x 1 cm, around the injection site) on the back of all SD rats on the 7th day after BTA injection. The wound size was measured and the expression of SP, CGRP, TGF-beta1 and alpha-SMA in wound granulation tissue was assayed by immunohistochemical staining and computerized image analysis before operation, and 3 days, 7 days and 14 days after operation.p><p>RESULTSAll the wounds healed 14 days after operation. The wound size in L and H group was not significantly different with that in C group on the 3rd day and 7th day after operation. The positive immuno-staining of SP, CGRP, TGF-beta1 and alpha-SMA in group L and H was significantly weaker than those in C group. Meanwhile, the positive immuno-staining of all above substances in H group was weaker than those in L group significantly.p><p>CONCLUSIONSBotulinum toxin type A can decrease the expression of SP, CGRP, TGF-beta1, and alpha-SMA in wound healing in a dose-dependent manner with no effect on the healing time.p>
Actins ; metabolism ; Animals ; Botulinum Toxins, Type A ; pharmacology ; Calcitonin Gene-Related Peptide ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Skin ; drug effects ; metabolism ; Substance P ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Wound Healing ; drug effects