牙髓炎症中降钙素基因相关肽阳性神经纤维的变化

本研究的目的在于探讨牙髓炎症及修复过程的分子生物学机理。在大鼠磨牙制备近髓窝洞，封入新鲜龋坏组织，建立了大鼠牙髓炎模型。用免疫组织化学方法观察正常牙髓及炎症４天、８天、１４天牙髓降钙素基因相关肽（ＣＧＲＰ）阳性神经纤维的变化。结果表明，ＣＧＲＰ阳性神经纤维广泛存在于正常牙髓中，大量阳性纤维进入牙本质小管；炎症４天组的牙髓中及进入牙本质小管内的阳性神经纤维均明显增多；炎症８天组冠髓部分坏死，阳性神经纤维包绕坏死区周围，根髓阳性神经纤维显著增多并可见大量“出芽”（ｓｐｒｏｕｔｉｎｇ）；炎症１４天组牙髓已大部分坏死，尖周膜腔阳性神经纤维聚集。结果提示，ＣＧＲＰ可能参与了牙髓炎症及其修复过程。     

正常与炎症牙髓中降钙素基因相关肽免疫反应神经纤维的研究

目的观察人正常与炎症牙髓中降钙素基因相关肽免疫反应神经纤维（CGRP－IRF）的分布，为深入探讨神经系统如何参与牙髓的各种生理，病理改变的调节作用提供新的资料。方法采用ABC免疫组化法观察人正常牙髓中CGRP－IRF的分布及龋源性炎症牙髓中CGRP－IRF的改变。结果牙髓中CGRP－IRF为许多串珠状棕褐色点状连成的线条，含明显的膨体。正常牙髓中，CGRP－IRF多沿血管走行并分布在血管周围，一些单根CGRP－IRF远离血管，主要分布在造牙本质细胞下神经丛。髓角处，偶尔可见CGRP-IRF突入造牙本质细胞层。炎症牙髓中，在病损下方和血管周围CGRP－IRF明显增多。结论CGRP－IRF广泛分布于人牙髓中，在牙髓炎症中起重要作用。     

不同发育阶段人牙髓中降钙素基因相关肽的变化

降钙素基因相关肽 (calcitoningene -relatedpeptide ,CGRP)是一种传递信息的多肽 ,几乎每一个生理、病理过程都有神经肽参与[1] 。免疫防御能力是在个体发生、发育过程中逐渐完善的 ,此过程中神经的变化可能反映了牙髓免疫防御功能完善过程。关于人年轻恒牙发育过程中牙髓CGRP含量与根尖闭合状态的关系 ,国内外未见相关报道。本研究采用免疫组织化学的方法 ,观察了CGRP数量变化及分布特征 ,为CGRP如何参与牙髓的生长发育提供依据。1　材料和方法牙髓标本取自因正畸需要而拔除的正常前磨牙 ,根尖未闭合组 15例 (10～ 12岁 ) ,根尖闭…     

人颞颌关节降钙素基因相关肽阳性神经纤维的分布

目的 :观察降钙素基因相关肽阳性神经纤维在人颞下颌关节的分布。方法 :利用免疫组化技术 ,观察正常成人 12人 ( 12侧 )颞下颌关节新鲜组织中 ,降钙素基因相关肽阳性神经纤维在关节的盘周附着和关节囊的内、外、前、后壁及关节盘中央的分布 ,并比较各个区神经纤维密度的异同。结果 :除关节盘中央外 ,关节囊及盘周组织均有丰富的降钙素基因相关肽阳性神经纤维 ,其中各个区密度分别是 :前部盘周附着和关节囊 ( 4 48.7± 78.5 )mm2 、后部 ( 4 2 6.7± 5 5 .9)mm2 、内侧 ( 2 0 5 .2± 3 9.9)mm2 、外侧 ( 2 95 .0± 3 6.6)mm2 。结论 :降钙素基因相关肽阳性神经纤维广泛分布于颞下颌关节软组织中 ,是感觉神经的一部分。     

正常与炎症牙髓中降钙素基因相关肽免疫反应神经纤维的研究

观察人正常与炎症攻髓中降钙素基因相关肽免疫反应神经纤维的分布，为深入探讨神经系统如何参与牙髓的各种生理，病理２的调节作用提供新的资料。方法采用ＡＢＣ免疫组化法观察人正常牙髓中ＣＧＲＰ－ＩＲＦ的分布及龋源性炎症牙髓中ＣＢＲＰ－ＩＲＦ的改变。     

咬合创伤去除前后牙髓组织中降钙素基因相关肽阳性神经纤维的变化

目的 研究咬合创伤对牙髓组织中降钙素基因相关肽(calcitonin gene-related peptide，CGRP)阳性神经纤维的影响，以及去除咬合创伤后牙髓组织中CGRP阳性纤维的恢复情况。方法 利用免疫组织化学方法，观察咬合创伤去除前后不同时期大鼠实验牙牙髓组织中CGRP阳性神经纤维的变化。结果 咬合创伤期实验牙牙髓组织中CGRP阳性纤维与对照牙相比逐渐出现数量增多、密度增加及纤维增粗的改变，而去除咬合创伤期实验牙牙髓组织中CGRP阳性纤维恢复接近正常大鼠。结论 咬合创伤能够引起大鼠牙髓组织中CGRP阳性纤维出现形态、分布、密度的改变，而去除创伤后大鼠牙髓组织中CGRP阳性纤维恢复接近正常大鼠。     

三叉神经痛患者神经纤维降钙素基因相关肽的观察

目的 :观察三叉神经痛患者痛支与非痛支神经纤维中降钙素基因相关肽的含量变化 ,加深对三叉神经痛发病机理的认识。方法 :用免疫组织化学法观察 16例患者痛支与非痛支神经纤维组织中降钙素基因相关肽免疫反应阳性颗粒的差异。结果 :发现痛支神经组织中降钙素基因相关肽免疫反应阳性颗粒的数量、面积均显著多于、大于非痛支神经组织中的降钙素基因相关肽免疫反应阳性颗粒。结论 :我们认为 :三叉神经痛的痛支神经过度合成和释放降钙素基因相关肽可能促进了SP的释放 ,导致阵发性剧烈疼痛 ,并在局部形成神经源性炎症。     

大鼠根尖周炎中降钙素基因相关肽阳性神经纤维的变化

目的：观察降钙素基因相关肽阳性(CGRP—IR)神经纤维在大鼠根尖周炎中的变化，为进一步研究CGRP在根尖周炎中的作用提供形态学依据。方法：在大鼠磨牙制备近髓窝洞，封入新鲜龋坏组织，建立大鼠根尖周炎模型。免疫组织化学技术观察正常根尖周膜和炎症14、21d根尖周组织CGRP-IR神经纤维的变化。结果：根尖周炎时，大鼠磨牙根尖周膜CGRP—IR神经纤维增多、增粗、染色加重，并围绕在坏死组织周围。结论：根尖周炎时，根尖周膜内CGRP增多，并佗于炎症的前沿区，CGRP可能参与了根火周组织炎症及修复过程。     

牙髓中的P物质,降钙素基因相关肽

神经肽(ncuropeptides)是体内传递信息的多肽,主要分布于神经组织,也分布于其它组织.同一种神经肽按其分布不同可能起递质(transmitter),调质(modulator) 或激素(hormone)样作用.近十多年来神经生物学领域中最重要的进展之一是对神经肽的研究.P物质(Substance P,SP)与降钙素基因相关肽(calcitoningene-related peptide,CGRP)同属神经肽类,其神经末稍中枢端止于脊髓背角,外周端止于内脏器官和皮肤粘膜等组织.SP和CGRP亦存在于大多数的口腔组织中,在牙髓中的含量相当高.最近的研究表明它们可能在牙髓的炎症和愈合过程中起重要的作用.     

降钙素基因相关肽和神经生长因子在损伤牙齿中的表达变化

目的：探讨降钙素基因相关肽（CGRP）和神经生长因子（NGF）在损伤牙齿中的表达变化及作用。方法：应用免疫组织化学方法，比较正常和机械损伤状态下CGRP和NGF在牙髓组织中的变化。结果：在正常牙髓组织中CGRP呈阳性反应，NGF染色呈阴性；损伤后即刻组CGRP染色弱阳性，NGF开始表达,3d组NGF牙髓染色强阳性；损伤5d后CGRP染色强阳性，NGF表达开始减弱；损伤9d后两者表达恢复到正常水平。结论：CGRP和NGF可能参与并促进牙齿损伤后的痛过敏和神经再生修复。     

The effect of orthodontic forces on calcitonin gene-related peptide expression in human dental pulp

Introduction

The purpose of this study was to quantify the effect of moderate and severe orthodontic forces on calcitonin gene-related peptide (CGRP) expression in healthy human dental pulp.

Methods

Thirty human dental pulp samples were obtained from healthy premolars in which extraction was indicated for orthodontic reasons. Before extraction, teeth were divided into 3 groups of 10 premolars each: (1) the control group: healthy premolars without application of orthodontic forces; (2) the moderate force group: a 56-g force was applied to the premolars for 24 hours; and (3) the severe force group: a 224-g force was applied to the premolars for 24 hours. All dental pulp samples were processed, and CGRP was measured by radioimmunoassay.

Results

Greater CGRP expression was found in the severe force group followed by the moderate force group. The lower CGRP values were for the control group. The Kruskal-Wallis test showed statistically significant differences between groups (P < .0001). Least significant difference (LSD) post hoc tests showed statistically significant differences in CGRP expression between the control group and the severe force group (P < .0001) but not with the moderate force group (P = .06). Differences between the moderate and severe force groups were statistically significant (P < .0001).

Conclusions

CGRP expression in human dental pulp increases when teeth are submitted to severe orthodontic forces.     

免疫荧光法观察正畸大鼠牙髓CGRP的变化

目的:观察正畸牙齿移动不同时期大鼠牙髓组织中降钙素基因相关肽(CGRP)阳性神经纤维的变化。方法:采用冷冻切片和间接免疫荧光方法观察正畸大鼠牙髓组织CGRP阳性神经纤维的表达变化情况。结果:CGRP阳性神经纤维在大鼠牙髓组织中分布较丰富,在根髓呈粗大的束状;在冠髓呈条索状、串珠样,并在髓顶造牙本质细胞下层,成网状分布。正畸加力后3d大鼠牙髓CGRP阳性神经纤维数量开始增加;加力7d达到最高;至撤力后28d下降到正常水平。结论:CGRP阳性神经纤维参与了正畸牙齿移动大鼠牙髓组织的反应过程。     

牙本质基质蛋白1在人不同龋坏牙齿中的免疫定位

目的 :观察牙本质基质蛋白 1(dentalmatrixprotein ,DMP1)在不同龋坏人牙齿中的表达 ,探讨DMP1在牙髓损伤修复中的作用。方法 :采用免疫组化SABC方法观察DMP1在不同龋坏人恒牙中的表达 ,并用图像分析系统进行半定量分析。结果 :DMP1在正常人恒牙组表达阴性 ,浅龋组部分成牙本质细胞和前期牙本质中弱阳性表达 ;中龋组成牙本质细胞数目增多 ,在胞浆和胞突以中阳性表达 ;深龋组成牙本质细胞层排列紊乱 ,出现空泡变性 ,成牙本质细胞样细胞形态细长、扁平 ,两种细胞胞浆和胞突中DMP1表达阳性 ,在修复性牙本质层强阳性表达。与对照组相比 ,成牙本质胞浆中DMP1含量在中龋组和深龋组明显增加 ,浅龋组无显著意义。结论 :DMP1参与成牙本质细胞样细胞分化分泌 ,并在修复性牙本质和反应性牙本质形成过程中起重要作用     

Neuropeptide Y Y1 receptor in human dental pulp cells of noncarious and carious teeth

Aim To determine the distribution of the NPY Y1 receptor in carious and noncarious human dental pulp tissue using immunohistochemistry. A subsidiary aim was to confirm the presence of the NPY Y1 protein product in membrane fractions of dental pulp tissue from carious and noncarious teeth using western blotting. Methodology Twenty two dental pulp samples were collected from carious and noncarious extracted teeth. Ten samples were processed for immunohistochemistry using a specific antibody to the NPY Y1 receptor. Twelve samples were used to obtain membrane extracts which were electrophoresed, blotted onto nitrocellulose and probed with NPY Y1 receptor antibody. Kruskal–Wallis one‐way analysis of variance was employed to test for overall statistical differences between NPY Y1 levels in noncarious, moderately carious and grossly carious teeth. Results Neuropeptide Y Y1 receptor immunoreactivity was detected on the walls of blood vessels in pulp tissue from noncarious teeth. In carious teeth NPY Y1 immunoreactvity was observed on nerve fibres, blood vessels and inflammatory cells. Western blotting indicated the presence and confirmed the variability of NPY Y1 receptor protein expression in solubilised membrane preparations of human dental pulp tissue from carious and noncarious teeth. Conclusions Neuropeptide Y Y1 is expressed in human dental pulp tissue with evidence of increased expression in carious compared with noncarious teeth, suggesting a role for NPY Y1 in modulation of caries induced pulpal inflammation.     

降钙素基因相关肽对人牙髓细胞的矿化影响

目的：分析降钙素基因相关肽（calcitonin gene-related peptide,CGRP)对人牙髓细胞促矿化功能的影响，探讨CGRP在牙髓炎症状态下表达量增多的生物学意义。方法：人牙髓细胞分实验组和对照组培养，实验组加入适量的CGRP。分别测定细胞的碱性磷酸酶活性、骨钙素含量、I型胶原mRNA表达量变化。结果：培养3、6、9、12、15d,5个时间点实验组比对照组细胞碱性磷酸酶活性增强、骨钙素含量增加，且有显著性差异（P＜0．01）。培养6d的实验组的细胞I型胶原mRNA表达比对照组明显高。结论：CGRP刺激牙髓细胞分化，促进矿化形成，CGRP表达增高将有助于修复性牙本质形成。     

Role of substance P and calcitonin gene-related peptide in the regulation of interleukin-8 and monocyte chemotactic protein-1 expression in human dental pulp

AIM: To determine whether leucocyte infiltration during neurogenic inflammation in the pulp is regulated by neuropeptides via inducing the release of proinflammatory chemokines interleukin-8 (IL-8) and monocyte chemotactic protein-1 (MCP-1) from human dental pulp. METHODOLOGY: Cultured primary pulp cells and pulp tissue explants were stimulated with substance P (SP) and/or calcitonin gene-related peptide (CGRP). IL-8 or MCP-1, secreted from cultured cells or produced in pulp explants, was analysed by enzyme-linked immunosorbent assay. RESULTS: Substance P induced IL-8 secretion from cultured pulp cells (approximately threefold increase over control, P < 0.05) and from pulp tissue explants (two- to three fold). SP only minimally to moderately induced MCP-1 (approximately two fold) in cultured pulp cells. While MCP-1 induction in cultured pulp cells was detected after 24 h of SP stimulation, no induction was observed in pulp tissue. CGRP did not induce IL-8, but moderately increased MCP-1 production (approximately three fold) in cultured pulp cells. There was no synergistic induction of MCP-1 by SP plus CGRP stimulation of pulp cells. CONCLUSIONS: Substance P is a stronger inducer of IL-8 production in dental pulp than CGRP. IL-8 is more strongly induced than MCP-1 by SP, suggesting a more important role for IL-8 than MCP-1 in leucocyte infiltration during neurogenic inflammation in dental pulp.     

Calcitonin gene-related peptide receptor expression in healthy and inflamed human pulp tissue

AIM: To use radioreceptor analysis for comparing calcitonin gene-related peptide (CGRP) receptor expression in human pulp tissue samples collected from teeth having a clinical diagnosis of acute irreversible pulpitis, healthy pulps and teeth with induced inflammation. METHODOLOGY: Six pulp samples were obtained from teeth having a clinical diagnosis of acute irreversible pulpitis. Another eight pulp samples were obtained from healthy premolars where extraction was indicated for orthodontic purposes. In four of these premolars, inflammation was induced prior to pulp collection. All the samples were processed and labelled with 125I-CGRP. Binding sites were identified by 125I-CGRP and standard CGRP competition assays. RESULTS: CGRP receptor expression was found in all human pulp tissue samples. Most receptors were found in the group of pulps from teeth having a clinical diagnosis of acute irreversible pulpitis, followed by the group of pulps having induced inflammation. The least number of receptors was expressed in the group of healthy pulps. The Kruskal-Wallis and Mann-Whitney (post-hoc) tests showed statistically significant differences between the groups (P < 0.05). CONCLUSION: CGRP receptor expression in human pulp tissue is significantly increased during inflammatory phenomena such as acute irreversible pulpitis.     

蟾毒对狗牙髓CGRP免疫阳性神经纤维的影响

目的：观察狗牙髓直接封蟾毒后牙髓内降钙素基因相关肽免疫反应阳性（简称ＣＧＲＰ－ＩＲ）神经纤维的动态变化。方法：采用冰冻切片和免疫组织化学方法（ＡＢＣ法）。结果：封药１ｈ，蟾毒组及可卡因组其牙髓内ＣＧＲＰ－ＩＲ神经纤维在形态、染色及数量上无显著差别；封药４ｈ，蟾毒组牙髓内ＣＧＲＰ－ＩＲ神经纤维明显断裂、数量减少、染色变浅，而可卡因组则无明显变化。结论：蟾毒快速麻痹牙髓神经的作用机理可能为一种神经毒性作用