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。     

大鼠磨牙钻磨后HSP70在牙髓不同时间表达的免疫组化研究

目的：研究大鼠磨牙受到钻磨刺激后HSP70在牙髓组织中的表达特点,推测其在牙髓损伤修复中的生物学作用。方法：采用免疫组织化学方法观察大鼠磨牙钻磨损伤后4h-14d内诱导型HSP70在牙髓组织中的动态表达及定位情况。结果：HSP70的表达在损伤早期呈强阳性,随后表达逐渐减弱,至修复期为弱阳性表达;同一时期受损牙髓的不同区域存在强弱变化。结论：HSP70大鼠磨牙受到钻磨损伤后呈现由强到弱的动态变化及区域性变化,这可能与牙髓损伤的应激保护和修复过程中蛋白合成的需要有关。     

下齿槽神经缺损神经移植后牙髓组织学实验研究

作者就下齿槽神经血管束缺损及神经移植后进行了牙髓组织学观察。通过对兔牙齿的电活力测验，牙髓微血管、组织学形态及牙髓神经纤维的实验观察，表明：牙齿电活力变化因不同时期神经血管束损伤而逐渐恢复。牙髓微血管在神经血管损伤初期，数量减少且扩张渗出；修复晚期，三级血管结构正常且丰富。牙髓组织细胞学变化早期呈不同程度的病理改变，后逐渐趋于正常。牙髓神经纤维也呈渐进性再生，由此认为，无论下齿槽神经血管束损伤修复     

牙髓损伤性刺激对牙髓组织中P物质等的影响

目的：比较和分析急性牙髓炎和牙齿开髓刺激后，牙髓组织中Ｐ物质（ＳＰ）、降钙素基因相关肽（ＣＧＲＰ）的变化。方法：利用免疫组织化学方法观察大鼠牙齿开髓后２ｈ、４ｈ和急性牙髓炎２ｈ、４ｈ组牙髓组织中ＳＰ、ＣＧＲＰ免疫阳性纤维的变化。结果：牙髓炎性及机械损伤性伤害刺激引起牙髓组织ＳＰ／ＣＧＲＰ免疫阳性纤维一致性变化。牙齿开髓及牙髓炎２ｈ、４ｈ组牙髓组织中ＳＰ、ＣＧＲＰ免疫阳性纤维减少。结论：两种刺激均引起牙髓组织内神经末梢释放ＳＰ、ＣＧＲＰ。     

大鼠磨牙牙髓组织在程度不同的机械性损伤刺激后HSP70的表达及意义的研究

目的 检测大鼠磨牙在程度不同的机械性钻磨损伤刺激(浅磨、深磨、穿髓)后热休克蛋白70(HSP70)的动态表达。方法 采用"二步法"免疫组化法观察HSP70在正常组、浅磨组、深磨24h组、穿髓24h组、穿髓3d组和穿髓7d组牙髓组织中的表达情况。结果 浅磨组和正常组HSP70在牙髓组织中有个别标本弱阳性着色,与后4组相比P<0.01;深磨24h组可见成牙本质细胞、成纤维细胞阳性表达,血管内皮细胞弱阳性表达;穿髓24h组和3d组表现为急性炎症反应,HSP70在成牙本质细胞、血管内皮细胞、中性粒细胞中均呈强阳性表达,成纤维细胞呈阳性表达;至穿髓7d时为弱阳性表达。结论 在程度不同的钻磨刺激下,HSP70在鼠牙髓组织中的表达有强弱不同的动态变化;HSP70在牙髓损伤修复过程中可能是主要介导牙齿矿化修复的细胞因子之一。     

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牙外伤包括牙体硬组织损伤、牙髓组织损伤和牙周组织损伤。牙髓损伤常见于牙齿折断、牙齿移位和牙齿全脱出中,可见牙髓损伤几乎在所有的牙齿外伤中都存在。外伤后,牙髓组织的转归可分为牙髓存活、牙髓钙变和牙髓坏死3种。牙髓组织的转归与以下因素有关:(1)外伤冲击力对牙髓组织的损伤;(2)外伤后外界不良刺激;(3)牙齿发育程度、个体差异等。儿童恒牙外伤后牙齿尚处于生长发育中,其治疗和预后远比成人复杂。本文针对年轻恒牙的特点,提出外伤后牙髓损伤判断和处置的对策。     

电子麻醉对牙髓SP、CGRP阳性纤维的作用

目的：研究电子麻醉镇痛作用的外周机理。方法：用免疫组化方法观察电子麻醉对大鼠牙髓及牙周组织感觉神经肽的影响。结果：电活力测试仪刺激可使被测牙牙髓内SP、CGRP阳性纤维完全脱失,牙周组织及领域牙不受影响。而电子麻醉使被累及的牙齿牙髓中的SP、CGRP阳性纤维数量减少,刺激部位牙龈组织中的SP、CGRP阳性纤维脱失。结论：电活力测试仪刺激仅兴奋牙髓中的感觉神经纤维,而电子麻醉可同时兴奋牙周组织及牙髓中的感觉神经纤维。电子麻醉和电活力测试仪刺激所兴奋的神经纤维可能属于不同的功能亚群,它们的兴奋分别产生镇痛作用的疼痛。     

Er:YAG激光备洞对牙髓P物质表达的影响

目的探讨Er:YAG激光备洞对牙髓的生物学效应。方法雄性SD大鼠66只随机分为实验组与对照组,实验组大鼠左上第一磨牙用高速牙钻备洞,右上第一磨牙用Er:YAG激光备洞,对照组大鼠磨牙不作任何处理。于不同时段分批处死大鼠,取标本做HE染色进行牙髓组织病理学观察,免疫组化法观察P物质(substance P,SP)的表达情况。结果 Er:YAG激光备洞后牙髓组织病理学改变及P物质的表达改变恢复正常要快于高速牙钻备洞。结论 Er:YAG激光备洞对牙髓的刺激小。     

牙髓诊断试验的临床应用

在牙体牙髓和根尖周疾病的诊治中．准确判断牙髓状态非常重要．它直接关系到对疾病的正确诊断、合理的治疗设计及对预后的准确估计。临床工作中对牙髓实施的冷测、热测和电测是将温度刺激或电刺激施加于牙齿硬组织上,牙髓组织中的A6神经纤维接受到刺激后会作出相应的反应．产生不同程度的痛感．据此．医生对牙髓状态做出判断。但此时患牙的反应仅指示其神经纤维的功能强度,并不能提供牙髓血流的信息．因此不能反映出牙髓的病理状态。近年来．发展出一些测定牙髓血供状态的新诊断方法．如激光多普勒血流测定仪、脉搏血氧饱和度测试仪．这些测试方法的结果更能真实地反映牙髓的病理状态．但仍需结合病史、临床检查、X线片检查来综合判断牙髓状况。本文通过回顾牙髓诊断试验的文献,对各种方法的原理．临床应用、可靠性及安全性进行综述．旨在为临床实践提供有益的参考。     

大鼠磨牙被钻磨后牙髓中诱导型热休克蛋白70 mRNA的表达

目的：探讨热休克蛋白70（HSP70）下牙髓自我保护的关系,以及HSP70表达是否存在的阈值。方法：将SD大鼠分为浅磨组、深磨组和对照组,采用原位杂交方法观察大鼠磨牙受到钻磨刺激后4h,HSP70mRNA是否表达以及在大鼠牙髓中的杂交定位。结果：HSP70在浅磨组与正常对照组牙髓中呈阴性,在深磨组牙髓中呈阳性。分布于成牙本质细胞、牙髓成纤维细胞和血管平滑肌细胞及内皮细胞。结论：牙髓组织受外界刺激时,可诱导HSP70表达。诱导型HSP70的表达可能存在阈值。进一步研究制洞深度与HSP70表达的相关性,将对临床操作有重要指导意义。     

The effect of capsaicin on substance P expression in pulp tissue inflammation

AIM: To evaluate the effect of capsaicin on substance P (SP) expression during induced inflammation in rat pulp tissue. METHODOLOGY: Radioimmunoanalysis was used to measure SP levels in 36 mandibular molar pulps taken from six Wistar rats. Twelve samples were obtained from healthy pulps and used as negative control group. Another 12 samples were obtained after inducing inflammation with mechanical pulp exposure; these were used as the positive control group. Capsaicin was infiltrated into the inferior dental nerve in the experimental group and 12 samples were obtained after mechanical pulp exposure. RESULTS: The lowest SP expression was found in mechanically exposed pulps where capsaicin pretreatment had been carried out (0.028 ng mL(-1)), followed by healthy pulps (0.302 ng mL(-1)). The highest SP expression was found in mechanically exposed pulps with no capsaicin pretreatment (124 ng mL(-1)). The Kruskal-Wallis test showed statistically significant differences between the groups (P < 0.001). CONCLUSION: Inferior dental nerve infiltration with capsaicin reduces SP expression in dental pulp tissue in rats.     

卫星胶质细胞活化在束缚应激致大鼠咬肌痛觉敏感中的作用

目的:观察慢性心理应激致咬肌痛觉敏感中三叉神经节卫星胶质细胞活化和P物质释放的关系。方法:40只健康雄性SD大鼠随机分为空白对照组、束缚应激3 d组、束缚应激7 d组、束缚应激14 d组。用体质量检测、旷场实验观察束缚应激后大鼠是否产生焦虑、抑郁样情绪;用Von Frey电子测痛仪检测大鼠咬肌痛阈。用免疫组织荧光染色法检测三叉神经节卫星胶质细胞活化的标志物神经胶质酸性蛋白(Glial fibrillary acidic protein, GFAP)及神经肽P物质(Substance P, SP)免疫阳性产物的变化。结果:受束缚应激后,大鼠体质量增长明显低于对照组,在旷场中总移动距离和总移动速度均明显降低,提示大鼠处于焦虑、抑郁样情绪状态。束缚应激后大鼠咬肌机械性痛阈明显降低;提示大鼠咬肌痛敏增强。束缚应激7 d、14 d组大鼠三叉神经节内GFAP及SP的表达明显高于空白对照组。结论:慢性心理应激可导致三叉神经节中的卫星胶质细胞活化,神经元SP表达升高。因此,三叉神经节中卫星胶质细胞和神经元的活化可能参与了慢性心理应激引起咬肌痛觉敏感升高过程。     

Substance P enhances the inhibition of osteoblastic cell differentiation induced by lipopolysaccharide from Porphyromonas gingivalis

BACKGROUND: Substance P (SP) is a multifunctional neuropeptide that transmits pain signals, regulates the immune system, and may modulate emotional stress. SP stimulates bone resorption activity of osteoclasts, and SP level in gingival crevicular fluid is correlated with the degree of periodontal inflammation. However, the exact roles of SP in bone metabolism and periodontal diseases are poorly understood. To elucidate the effect of stress on bone metabolism, we investigated the effect of SP on osteoblastic cell differentiation in the presence of lipopolysaccharide from Porphyromonas gingivalis (P-LPS). METHODS: The primary osteoblastic cells were isolated from fetal rat calvaria (RC) and cultured with SP, P-LPS, and an SP antagonist (SPa). The effects of SP on bone nodule (BN), alkaline phosphatase (ALPase) activity, mRNA expressions of SP receptor, bone matrix proteins, and Cbfa 1 were investigated. RESULTS: SP stimulated the expression of SP receptor mRNA in RC cells and enhanced its expression in the presence of P-LPS (50 ng/ml). SP inhibited BN formation and ALPase activity in a dose-dependent manner (10(-7) to 10(-5) M) and further suppressed mRNA expression of bone sialoprotein, osteopontin, and osteocalcin but not of type I collagen mRNA. The inhibitory effects were enhanced in the presence of P-LPS and blocked by Spantide III. Furthermore, the expression of Cbfa 1 mRNA was also markedly suppressed in the presence of SP and P-LPS. CONCLUSION: These findings suggest that SP inhibits osteoblastic cell differentiation and may be related to bone metabolism in periodontal diseases under conditions of stress.     

Quantification of neuropeptides (calcitonin gene-related peptide, substance P, neurokinin A, neuropeptide Y and vasoactive intestinal polypeptide) expressed in healthy and inflamed human dental pulp

AIM: To quantify the expression of calcitonin gene-related peptide (CGRP), substance P (SP), neurokinin A (NKA), neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP) in healthy and inflamed human dental pulp tissue. METHODOLOGY: Six pulp samples were obtained from teeth having a clinical diagnosis of acute irreversible pulpitis. Another 12 pulp samples were obtained from premolars where extraction was indicated for orthodontic purposes. In six of these premolar teeth inflammation was induced by mechanical pulp exposure prior to sample collection. All samples were processed and 125I-labelled; neuropeptides were quantified by competition assays. ANOVA and Mann-Whitney's (post hoc) tests were used to establish statistically significant differences between the groups. RESULTS: Expression of five neuropeptides was found in all human pulp samples. Statistical analysis revealed a significantly higher (P < 0.05) expression of CGRP, SP, NKA and NPY in both inflammatory conditions compared with healthy pulp control values. VIP expression remained stable during the inflammatory conditions. CONCLUSION: Expression of CGRP, SP and NKA released from C-fibres and NPY released from sympathetic fibres is significantly higher in the inflamed human pulp compared with healthy pulp. Expression of VIP released from parasympathetic fibres is not increased during the inflammatory conditions of human dental pulp.     

龈沟液中神经肽P物质与慢性牙周炎关系临床研究

目的:探讨龈沟液中神经肽P物质(SP)与慢性牙周炎发生发展的关系.方法:研究组48 例,对照组45 例,研究组口内各选1 个牙周炎牙位、对照组各选1 个健康牙位收集龈沟液样本,采用放射免疫分析技术分别测定2 组龈沟液SP含量;记录牙龈指数(GI)、牙周袋探诊深度(PD)、附着丧失(AL)、牙槽骨吸收(ABL)情况,并进行与SP的相关性分析.结果:研究组SP含量显著高于对照组(t＝9.92,P<0.01);轻度牙周炎SP含量及牙周临床指标与对照组相比,均有显著差异(P<0.01);轻度牙周炎龈沟液SP含量与GI无显著相关性,但与PD、AL、ABL之间,以及中、重度牙周炎龈沟液SP含量与GI之间,均呈显著性相关(P＜0.05);中、重度牙周炎SP含量与PD、AL、ABL之间均呈非常显著性相关(P<0.01).结论:龈沟液中SP含量与慢性牙周炎的发生发展密切相关,可以反映牙周组织的破坏程度;测定患者龈沟液中SP含量,对于判断病情、指导治疗具有重要意义.     

iNOS在口腔扁平苔藓癌变过程中的表达研究

目的 :研究诱导型一氧化氮合酶 (iNOS)在口腔扁平苔藓、口腔鳞状细胞癌发展过程中的表达水平及其作用。方法 :以正常口腔黏膜作对照 ,采用免疫组化SP法检测 2 5例口腔扁平苔藓和 10例口腔鳞状细胞癌组织中的iNOS表达。结果 :正常口腔黏膜iNOS阴性表达 ;扁平苔藓组和鳞癌组iNOS表达较正常黏膜组均非常显著增加 (P <0 .0 1) ;扁平苔藓组与鳞癌组之间以及扁平苔藓组内部按上皮异常增生程度分组之间iNOS的表达均无显著差异 (P >0 .0 5 )。结论 :iNOS在口腔扁平苔藓伴上皮异常增生和鳞癌中高表达 ,在口腔鳞癌的发生发展中起着重要的作用。     

Heme oxygenase‐1 protects human periodontal ligament cells against substance P‐induced RANKL expression

Lee H‐J, Jeong G‐S, Pi S‐H, Lee S‐I, Bae W‐J, Kim S‐J, Lee S‐K, Kim E‐C. Heme oxygenase‐1 protects human periodontal ligament cells against substance P‐induced RANKL expression. J Periodont Res 2010; 45: 367–374. © 2010 John Wiley & Sons A/S Background and Objective: Although substance P (SP) stimulates bone resorption activity and this is reported to be correlated with the degree of periodontal inflammation, it is unclear how human periodontal ligament cells regulate neuropeptide‐induced osteoclastogenesis or the possible involvement of heme oxygenase‐1 (HO‐1) might be. This study examines how SP affects osteoprotegerin (OPG) and RANKL expression via HO‐1. Material and Methods: Using immortalized human periodontal ligament cells, the effects of SP on the expression of HO‐1, RANKL and OPG mRNA and proteins were determined by RT‐PCR and western blotting, respectively. Various concentrations of SP (10^?7, 10^?8, 10^?9 and 10^?10 m ) were added to the medium, and the cells were treated for 0, 0.25, 0.5, 1, 2 and 3 d. Results: Substance P upregulated RANKL and HO‐1 and downregulated OPG mRNA and protein expression in periodontal ligament cells, in a concentration‐ and time‐dependent manner. A HO‐1 inducer inhibited both the upregulation of RANKL expression and downregulation of OPG expression by SP in periodontal ligament cells. By contrast, treatment with a HO‐1 inhibitor or HO‐1 small interferring RNA (siRNA) enhanced SP‐stimulated RANKL expression. Inhibitors of ERK and p38 MAP kinases, phosphoinositide 3‐kinase and nuclear factor‐κB blocked the effects of SP on RANKL expression in periodontal ligament cells. Conclusion: These results suggest that SP stimulates osteoclastic differentiation by increasing the expression of RANKL vs. OPG via the HO‐1 pathway in periodontal ligament cells. The HO‐1 pathway may be an effective therapeutic target for inhibiting chronic periodontitis involving alveolar bone resorption.     

人正常牙髓和炎症牙髓中P物质和血管内皮生长因子的表达

目的:研究P物质(SP)和血管内皮生长因子(VEGF)在人正常、外伤初期、炎症早期、炎症晚期牙髓中的表达和相关性,探讨两者对牙髓微循环的调节作用。方法:选取因正畸或阻生需拔除的健康牙10颗作为正常组,选取同年龄阶段因外伤需要根管治疗的患牙29颗,分为外伤初期组、早期炎症组、晚期炎症组。采用免疫组织化学方法对各组中SP、VEGF进行组织学定位和定量检测。采用SSPS13.0软件包对各组数据进行单因素方差分析和相关分析。结果:与正常组相比,SP在其他各组中表达显著降低(P<0.05);VEGF在早期炎症组中表达较其他各组强(P<0.05),而在晚期炎症组则明显减弱(P<0.05);SP和VEGF在各组中的表达呈负相关(r=-0.378,P<0.05)。结论:SP可能通过VEGF途径引起牙髓微循环的病理改变,两者共同参与牙髓神经源性炎症的发生。     

人端粒酶逆转录酶与c-myc和刺激蛋白1在成釉细胞瘤中的表达

目的　研究原癌基因转录因子c-myc和刺激蛋白1(SP1)在人成釉细胞瘤(AB)中的表达及与端粒酶逆转 录酶(hTERT)的关系。方法　选取54例AB(原发31例,复发19例,恶变4例)存档蜡块,采用原位杂交检测AB中 c-myc mRNA、hTERTmRNA的表达,采用免疫组化SP法,检测AB中SP1蛋白的表达。结果　c-myc mRNA在AB中 阳性率为81·5%(44/54),hTERTmRNA为94·4%(51/54),SP1为83·3%(45/54);伴随AB的复发与恶变,3者表达强 度上升;相关性分析得出hTERT与c-myc之间存在相关性(rs=0·853,P<0·001),hTERT与SP1之间存在相关 性(rs=0·900,P<0·001)。结论　hTERT在AB中明显增加的活性可能与转录因子c-myc和SP1有关,3者在AB中 的表达趋势与AB的临床生物学特性相关。