Objective To investigate the significance of sensory neuropeptides [calcitonin gene related peptide (CGRP) and substance P (SP)] in steroid-induced avascular necrosis of the femoral head (ANFH) by using a rabbit model. Methods Fifty-five adult female Japanese White rabbits (weighing 3 kg and aging 24 months) were randomly divided into experimental group (n=45) and control group (n=10). The rabbits in experimental group received a single intramuscularinjection of methylprednisolone at a dose of 4 mg/kg and then were sacrificed after 3 days (n=15), 1 week (n=15), and 2 weeks (n=15) of injection. The rabbits in control group were fed without any treatment. The necrosis of the femoral head was observed. And the expressions of the monoclonal antibodies CGRP and SP were observed with immunohistochemical staining. Also, the integrated absorbance (IA) value of the positive area was calculated. Results All rabbits survived to the end of the experiment. There was no necrosis of the bone or bone marrow in experimental group at 3 days; whereas ANFH was observed in 5 rabbits at 1 week (33%) and in 8 rabbits at 2 weeks (53%). There were significant differences in the rate of ANFH between 1 week, 2 weeks and 3 days (P lt; 0.05); but there was no significant difference between 1 week and 2 weeks (P gt; 0.05). The intensity of CGRP immunoreactivity increased and reached the peak at 1 week, and then decreased at 2 weeks in experimental group. The IA value of CGRP in experimental group at 1 week was significantly higher than that of control group and that of experimental group at 3 days (P lt; 0.05). The IA value of CGRP in experimental group at 2 weeks was significantly lower than those at 3 days and 1 week (P lt; 0.05). The intensity of SP immunoreactivity decreased and reached the lowest at 1 week, and then increased. The IA value of SP in experimental group at 1 week was significantly lower than that of control group and that of experimental group at 2 weeks (P lt; 0.05). Conclusion The sensory neuropeptides may be affected by the steroid, which may play a key role in the process of steroid-induced ANFH by imbalance of bone metabol ism, disturbance of the microcirculation of bone, and disorder of the protective pain-transmission.
Objective To explore the effects of calcitonin gene-related peptide (CGRP) on the migration of bone marrow mesenchymal stem cells (BMSCs) and vascular endothel ial growth factor (VEGF) expression in vitro. Methods TheBMSCs were isolated from Sprague Dawley rats using whole bone marrow adherence method. At 1, 2, and 3 weeks after culture, the expressions of CGRP receptor (CGRPR) was detected by Western blot. The BMSCs were treated with CGRP at concentration 1 × 10-8 mol/L (experimental group) and did not treated (control group), and the efficacy of BMSCs migration was analyzed by Transwell chamber assay after 72 hours; at 1, 3, 5, and 7 days, the mRNA expressions of vascular cell adhesion molecule 1 (VCAM-1) were detected by real-time fluorescent quantitative PCR; the protein expressions of VEGF were examined using immunohistochemistry and Western blot. Results CGRPR expressed stably in the cultured BMSCs and reached the peak at 2 weeks. CGRP had a significantly enhanced role in promoting cell migration. The number of cell migration was (3.20 ± 1.77) cells/HP in experimental group and (1.11 ± 0.49) cells/HP in control group, showing significant difference (t=4.230, P=0.001). In experimental group, the expressions of VCAM-1 mRNA increased with time and reached the peak at 7 days. There were significant differences in the expressions of VCAM-1 mRNA between control group and experimental group at 3, 5, and 7 days (P lt; 0.05). Immunocytochemistry results showed positive DAB staining for VEGF at 5 and 7 days in experimental group. Western blot results showed that the protein expressions of VEGF increased significantly at 5 and 7 days in experimental group when compared with control group (P lt; 0.05), which was signfiantly higher at 5 days than at 7 days in experimental group (P lt; 0.05). Conclusion CGRP can promote the migration of BMSCs and stimulate the protein expression of VEGF, which may plays an important role in regulating bone metabol ism by increasing angiogenesis.
Objective To investigate the effect of curcumin on calcitionin gene related peptide (CGRP) expression after spinal cord injury (SCI) in rats. Methods A total of 200 rats, weighing 250-300 g, were randomly divided into 4 groups (n=50): sham-operation group, normal saline (NS) group, low-dose curcumin group (30 mg/kg), and high-dose curcumin group (100 mg/kg). In sham-operation group, only vertebral lamina excision was performed without SCI; the SCI model was established in the other 3 groups. At immediate after modeling, 30 mg/kg and 100 mg/kg curcumin were injected intraperitoneally in 2 curcumin groups, equivalent NS was given in NS group (30 mg/kg), but no treatment in sham-operation group. At 1, 3, 7, 14, and 21 days after operation, the motor neural function was evaluated by the inclined plane test and Basso-Beattie-Bresnahan (BBB) scores; immunohistochemical staining and Western blot assay were used to observe CGRP expression. Results BBB score and inclined plane test score of NS group, low-dose curcumin group, and high-dose curcumin group were significantly lower than those of sham-operation group at each time point (P lt; 0.05). BBB score of low-dose curcumin group and high-dose curcumin group was significantly higher than that of NS group at 3, 7, 14, and 21 days after SCI (P lt; 0.05), and the score of high-dose group was significantly higher than that of low-dose curcumin group at 7, 14, and 21 days after SCI (P lt; 0.05). Inclined plane test score of low-dose curcumin group and high-dose curcumin group was significantly higher than that of NS group at 7, 14, and 21 days after SCI (P lt; 0.05), and the score of high-dose curcumin group was significantly higher than that of low-dose curcumin group at 7, 14, and 21 days after SCI (P lt; 0.05). Immunohistochemical staining results showed that the CGRP positive cells of sham-operation group was significantly more than those of the other 3 groups, and the CGRP positive cells of high-dose curcumin group were significantly more than those of low-dose curcumin group at each time point (P lt; 0.05); the CGRP positive cells of low- and high-dose curcumin groups were significantly more than those of NS group at 3, 7, 14, and 21 days after SCI (P lt; 0.05). Western blot assay results showed that the CGRP protein expressed at each time point after SCI in sham-operation group; the CGRP protein expression gradually decrease with time passing in NS group; but the CGRP protein expression gradually increased with time passing in low- and high-dose curcumin groups, and reached the peak at 14 days, then maintained a high level. Conclusion After SCI in rats, 30 mg/kg curcumin can improve rats’ motor function, and 100 mg/kg curcumin effect is more obvious, especially in promoting the expression of CGRP. That may be the mechanism of protection of the nervous system.
Objective To explore the changes of calcitonin gene-related peptide (CGRP) and substance P (SP) levels after end-toend and end-to-side neurorrhaphy. Methods Twenty female Wistar rats were divided into 4 experimental groups and control group. In the experimental groups, common peroneal nerves were transected on both sides. End-to-side coaptation was performed on the left, while end-to-end coaptation on the right. After 1, 2, 4 and 27 weeks, the rats were sacrificed, and immunoreactivities of CGRP and SP in suture sites, lumbar spine and dorsal root ganglia(DRGs) were evaluated respectively. Results The expression ofCGRP and SP decreased in dorsal horn and DRGs within 1 week postoperatively. After 4 -27 weeks, CGRP and SP in dorsal horn could return to almost normal level, but they had little recovery in DRGs. Although the trend of change between end-to-end and end-to-side was coincident, in most experimental groups, thereexisted differences in the dorsal horn between end-to-end and end-to-side. The sciatic nerve stained by acetylcholinesterase, SP, CGRP and PGP 9.5 showed that the fibers could pass through the suture site of either end-to-end or end-to-side. Conclusion Nerve regeneration can be achieved by end-to-side neurorrhaphy, andthe mechanism of sensory nerve recovery of these two methods is similar. But the recovery in end-to-side coaptation is insufficient to some degree.
【摘要】 目的 研究大剂量辣椒素对兔肺P物质和降钙素基因相关肽(CGRP)的影响。方法 将16只成年健康雄性新西兰大白兔随机分为两组:载体组(A)、大剂量辣椒素组(B)。两组动物取样前7 d和前6 d分别于颈部皮下注射等量的载体或者辣椒素(20 mg/mL)。大剂量戊巴比妥钠静脉麻醉处死动物后立即切取左肺下叶,称重,匀浆,离心后取上清液置于-80℃冰箱保存待测P物质和CGRP。结果 A组肺组织中P物质的浓度高于B组(Plt;0.05),而CGRP的浓度两组差异无统计学意义(P＞0.05)。结论 大剂量辣椒素(100 mg/kg)不能完全耗竭兔肺初级感觉神经纤维的神经肽类物质。
【摘要】 目的 研究降钙素基因相关肽（calcitonin gene related peptide， CRGP）在肝硬化门静脉高压症患者食管下段胃底静脉曲张中的作用。 方法 以2005年1月－2010年8月46例肝硬化门静脉高压症不同程度食管下段胃底静脉曲张患者作为研究对象，并按食管下段胃底静脉曲张严重程度分为轻度曲张组、中度曲张组、重度曲张组，以30例行胃肠疾病手术无肝病患者作为对照。术中水柱法测定门静脉压力；酶联免疫吸附法测定门静脉血中CGRP含量。 结果 对照组及轻、中、重度曲张组门静脉压力分别为（14.8±2.1）、（30.5±2.5）、（44.3±3.2）、（47.6±3.8） cm H2O（1 cm H2O=0.098 kPa）。门静脉血中CGRP的含量分别为（45.4±5.4）、（69.2±7.2）、（93.6±8.7）、（98.2±9.4） pg/mL。对照组门静脉压力及CGRP含量明显低于其他3组（Plt;0.05），在轻度曲张组明显低于中度和重度曲张组（Plt;0.05），中度和重度曲张组之间差异无统计学意义（Pgt;0.05）。 结论 CRGP在肝硬化门静脉高压症食管下段胃底静脉曲张的发生和发展中起重要作用，CGRP可作为反映食管静脉曲张程度的一种有用指标。【Abstract】 Objective To investigate the role of calcitonin gene related peptide (CRGP) in pathogenesis of esophageal varices in portal hypertension with cirrhosis. Methods from January 2005 to August 2010, 46 patients with portal hypertension and cirrhosis at different degrees of esophageal varices were divided into mild varicose group, moderate varicose group and severe varicose group according to the severity of esophageal varices. The patients who underwent gastrointestinal surgery without liver disease were as the control. Portal vein pressure was detected by mercury during the surgery. The expression of CGRP was assayed by enzyme-linked immunosorbent assay. Results The portal pressure was (14.8±2.1), (30.5±2.5), (44.3±3.2), and (47.6±3.8) cm H2O (1 cm H2O=0.098 kPa) in the control group and the mild, moderate and severe varicose group, respectively. Those CGRP content in the portal vein was (45.4±5.4), (69.2±7.2), (93.6±8.7), and (98.2±9.4) pg/mL, respectively. CGRP content and portal vein pressure were the lowest in control group, which were significantly lower than those in the other three groups (Plt;0.05); which were also significantly lower in mild varicose group than those in the moderate and severe esophageal varices group (Plt;0.05), while no statistic difference between moderate and severe esophageal varices group was found (Plt;0.05). Conclusion CGRP plays an important role in the occurrence and development of portal hypertension with cirrhosis concurrent esophageal varices, and it may serve as a useful indicator reflecting the degree of esophageal varices.
ObjectiveTo summarize the research progress on the calcitonin gene-related peptide (CGRP) and receptor activator of nuclear factor κB (RANK)/receptor activator of nuclear factor κB ligand (RANKL)/osteoprotegerin (OPG) system during bone reconstruction to provide theoretical basis for further research on the prevention and treatment of bone-related diseases.MethodsThe relevant research results at home and abroad in recent years were analyzed and summarized.ResultsCGRP and RANK/RANKL/OPG system play important regulatory roles in the bone reconstruction.ConclusionAt present, the research on the mechanism of CGRP and RANK/RANKL/OPG system in bone reconstruction is insufficient. Therefore, it is necessary to study further on the process and interrelation of CGRP and RANK/RANKL/OPG system in bone reconstruction to confirm their mechanism, which will bring new ideas and methods for the treatment of bone related diseases in clinic.
This study aims to investigate the effect of lung ischemia reperfusion injury (LIRI) on expression of transient receptor potential vanilloid 1 (TRPV1) in the lung and brainstem of rats. Sixteen adult male Sprague Dawley rats weighing 250-320 g were randomly divided into Sham group and ischemia reperfusion group (IR group). Before ischemia, 0.5 hour and 4 hours after the reperfusion, respectively, arterial partial pressure of oxygen (PaO2) and arterial-alveolar oxygen pressure gradient (A-aDO2) were recorded and calculated, respectively. Left lung tissues and the brainstems were obtained at the end of the experiment. Lung tissue malondialdehyde (MDA), myeloperoxidase (MPO) activities, calcitonin gene related peptide (CGRP) and substance P (SP) levels were assessed. The mRNA and protein expressions of TRPV1 in the lung and brainstem were measured by qRT-PCR and Western blot. Compared with in the Sham group, rats in the IR group had a poorer blood gas exchange (P<0.05) and the MPO activity and MDA level of lung tissues in the IR group were significantly higher than those in the Sham group (P<0.05). CGRP level in the IR group increased remarkably (P<0.05), while SP level did not differ statistically between the two groups (P>0.05). The mRNA and protein expressions of TRPV1 in the lung tissue were upregulated in the IR group (P<0.05), but there were no differences of those in the brainstem between the two groups (P>0.05). The results suggest that LIRI could upregulate the expressions of TRPV1 and evoke CGRP release in the lung.
Objective To investigate the effect and mechanism of calcitonin gene-related peptide (CGRP) on the prevention and treatment of transplant vein graft disease. Methods The 25 New Zealand white rabbits were divided into three groups: an experimental group [n=8, the rabbit jugular veins transfected with adeno-associated virus vector tipe 2/1 containing CGRP gene (AAV2/1-CGRP)], a carrier group [n=9, transfected with mosaic adeno-associated virus vector tipe 2/1 containing LacZ gene (AAV2/1-LacZ)] and a control group (n=8, saline) and then the cervical veins were implantated into the ipsilateral carotid artery by reverse end-side anastomosis. At 4 weeks after surgery, the pathology of the specimens, CD68 immunohistochemistry, in situ β-galactosidase staining were obtained. The expression of CGRP gene was detected by reverse transcription-polymerase chain reaction (RT-PCR). Monocyte chemoattractant protein-1(MCP-1), tumour necrosis factor-α (TNF-α), inducible nitric oxide synthase (iNOS) and matrix metalloproteinase-9 (MMP-9) were detected by real-time polymerase chain reaction (real-time PCR). Results The CGRP and LacZ gene expression was positive at postoperative 4 weeks. The intima/media ratio was significantly inhibited in the experimental group. Macrophage infiltration and expression of inflammatory mediators including MCP-1, TNF-α, iNOS and MMP-9 were also significantly inhibited in the experimental group. Conclusion Transfection of AAV2/1-CGRP inhibits inflammatory mediator expression, macrophage infiltration and neointimal hyperplasia in experimental vein graft disease.
To observe the change of morphology and neuropeptide in the spinal neurons in order to clarify the functional state after injury of peripheral nerves is especially in the late stage. Sciatic nerves were cut with their proximal segments in the preparation of a model of peripheral nerve injury. Combination of horseradish peroxidase retrograde tracing immunohistochemistry and computer image analysis the changes in the morphometry of the perikarya of ventral horn neurons of the spinal cord, the quantitative changes of substance P (SP). Calcitonin gene-related peptide (CGRP) in dorsal horn and CGRP and choline acetyransferase (CHAT) in ventral horn of the spinal cord were examed. The results showd: (1) At the 3rd week after injury, swollen perikarya of the ventral horn neurons were observed, subseauently the swelling of perikarya was decreased tile the 6th week the neurons recovered to their normal size. At the 12th week the neurons were generally stable in their size, shortening of the dendrites was seen in 27% of the neurons. (2) The dendrites of the neurons progressively contracted till at the 12th week 53% of them were degenerated. The results of the 24th week were similar to the that at the 12th week. (3) CGRP in the ventral horn of the spinal cord was elevated to the highest point after 1 week of injury, that lasting for 4 weeks and 8 weeks later, the lever of CGRP returned to normal. From 20th to 24th week, there was no obvious changes of CHAT in the ventral horn of the spinal cord during observation. (4) SP went to the lowest point in the dorsal horn during 2-6 weeks, then recovered slowly, and beiny normal again after 16 weeks, however, CGRP was changed slightly. The results indicated that although a series of degenerating changes occurred in the neurons of the spinal cord during the late peripheral nerve injury, but the functional activity of the central meurons still was maintained at a certain level.