红景天抗大鼠嗅球衰老机制的研究

目的 :探讨红景天抗老龄大鼠嗅球衰老的机制。方法 :取健康大鼠 ,青龄组 (3个月 ) 10只 ;老龄组(2 6个月 ) 2 0只 (随机分成老龄对照组、老龄红景天组各 10只 ) ;均断头处死 ,取嗅球 ,经石蜡包埋 ,连续切片 ,免疫组化染色 ,光镜观察。结果 :成纤维细胞生长因子 (FGF)及凋亡相关基因 (Bcl 2和Bax)主要表达在大鼠嗅球的僧帽细胞 ;青龄组FGF和Bcl 2阳性细胞表达率均显著高于老龄对照组 (P <0 .0 1) ,且FGF和Bcl 2蛋白的表达呈明显正相关 (P <0 .0 1) ;而青龄组Bax阳性细胞表达率略低于老龄对照组 ,但差异不显著 (P >0 .0 5 )。老龄红景天组FGF和Bcl 2蛋白阳性细胞表达率明显高于老龄对照组 (P <0 .0 1)。结论 :大鼠嗅球老化与FGF和Bcl 2蛋白表达减少密切相关 ;推测红景天可能是通过上调嗅球中FGF蛋白的表达 ,诱导僧帽细胞Bcl 2蛋白表达增多 ,抑制嗅球中僧帽细胞的凋亡是其抗大鼠嗅球衰老的重要机制。     

大鼠嗅球中凋亡相关基因蛋白的表达及其意义

目的 :明确凋亡相关基因 Bcl- 2和 Bax在大鼠嗅球中的表达及分布 ,并探讨其意义。方法 :取健康大鼠青龄组 (3～ 6个月 ) ,成龄组 (12～ 15个月 ) ,老龄组 (2 4～ 2 6个月 )各 10只 ,将其断头处死 ,取嗅球 ,经石蜡包埋 ,连续切片 ,免疫组化染色 ,光镜观察。结果 :Bcl- 2和 Bax主要表达于大鼠嗅球中的僧帽细胞。Bcl- 2阳性细胞表达率青龄组显著高于老龄组 (P>0 .0 1) ;Bax阳性细胞表达率老龄组虽略高于青龄组 ,但差异不显著 (P>0 .0 5)。结论 :随着年龄的增长 ,嗅球中抗凋亡基因 Bcl- 2蛋白表达显著减少 ,提示凋亡与嗅觉老化的密切关系。推测嗅球中僧帽细胞的凋亡可能是老化性嗅觉障碍的重要因素之一。     

大鼠嗅球凋亡基因蛋白的表达与蛋白激酶C调控的探讨

目的:明确蛋白激酶Ｃ和凋亡相关基因Ｂｃｌ－２和Ｂａｘ在大鼠嗅球中的表达分布及相关性,探讨蛋白激酶Ｃ(ｐｒｏｔｅｉｎ ｋｉｎａｓｅ Ｃ,ＰＫＣ)对大鼠嗅球神经元凋亡的调节作用。方法:取青龄大鼠(４~６个月龄),成龄大鼠(１０~１２月龄);老龄大鼠(２４~２６月龄)各１０只,断头处死,将大鼠嗅球置于４%的多聚甲醛中固定,连续切片,应用免疫组化方法检测ＰＫＣ与凋亡基因Ｂｃｌ－２和Ｂａｘ在大鼠嗅球中的表达。结果:大鼠嗅球中的ＰＫＣ表达老龄组(７０．４±５．３８)明显高于青龄组(２０．４４±１．２１);而Ｂｃｌ－２的表达青龄组(７０．１６±５．０１)明显高于老龄组(２４．１０±４．００)。差异显著(Ｐ＜０．０１);且ＰＫＣ和Ｂｃｌ－２的表达呈负相关(ｒ＝０．８３７１)。Ｂａｘ随增龄逐渐增多,但差异不显著。结论:大鼠嗅球细胞的凋亡与ＰＫＣ的调节密切相关;推测ＰＫＣ对大鼠嗅球的生长、发育起着重要的负反馈调节作用。     

在鼠嗅球中凋亡相关基因蛋白的表达及其意义

目的：明确凋亡相关基因Ｂｃｌ－２和Ｂａｘ在大鼠嗅球中的表达及分布，并探讨其意义。方法：聚健康大鼠青龄组（３￣６个月），成龄组（１２￣１５个月），老龄组（２４￣２６个月）各１０只，将其断头处死，取嗅球，经石蜡包埋，连续切片，免疫组化染色，光镜观察。结果：Ｂｃｌ－２和Ｂａｘ主要表达于大鼠球中的僧帽细胞。Ｂｃｌ－２阳性细胞表达率青龄组显著高于老龄组（Ｐ〉０．０１）；Ｂａｘ阳性细胞表达率老龄组虽略高于青龄组，但差异不显著（Ｐ〉０．０５）。结论：随着年龄的增长，嗅球中抗凋亡基因Ｂｃｌ－２蛋白表达显著减少，提示凋亡与嗅觉老化的密切关系。推测嗅球中僧帽细胞的凋亡可能是老化性嗅觉障碍的重要因素之一。     

鼻科学

”1769成纤维细胞生长因子在大鼠嗅球中表达及其与老化性嗅觉减退的关系/关桂梅…//耳鼻咽喉一头颈外科一1999,6(2)一118一1 20,1 23 为探讨成纤维细胞生长因子(fi broblastgrowth factor,F(汗)在嗅球中表达及其与老化性嗅觉减退的关系,取健康幼龄组和老龄组大鼠各10只,将其断头处死,分别取嗅粘膜和嗅球,经石蜡包埋,连续切片,免疫组化染色,光镜观察。结果,幼龄组嗅球FGF阳性表达(100环)明显高于老龄组(30%)(P<0.01),而两组嗅粘膜均未见表达。结论,随着年龄的增长,嗅球中FGF表达明显降低,推测嗅球中FGF降低与老化性嗅觉减退有密切关系。…     

成纤维细胞生长因子在大鼠嗅球中表达及其与老化性嗅觉减退的关系

为探讨成纤维细胞生长因子（fibroblastgrowthfactor，FGF）在嗅球中表达及其与老化性嗅觉减退的关系，取健康幼龄组和老龄组大鼠各10只，将其断头处死，分别取嗅粘膜和嗅球，经石蜡包埋，连续切片，免疫组化染色，光镜观察。结果，幼龄组嗅球FGF阳性表达（100％）明显高于老龄组（30％）（P＜0．01），而两组嗅粘膜均未见表达。结论，随着年龄的增长，嗅球中FGF表达明显降低，推测嗅球中FGF降低与老化性嗅觉减退有密切关系。     

红景天素上调大鼠嗅球成纤维细胞生长因子及其受体机制的探讨

目的：明确大鼠嗅球中成纤维细胞生长因子（FGF）合成、分泌和作用的靶细胞及其与受体（FGFR）表达的相关性；探讨中药红景天素抗大鼠嗅球衰老的信号传导机制。方法：取Wister大鼠青年对照组；老龄对照组和老龄红景天素用药组各10只，3个月后断头处死，取嗅球，以原位杂交和免疫组化方法分别检测FGFmRNA和FGF、FGFR蛋白在大鼠嗅球中的表达。结果：随着年龄的增长，大鼠嗅球中FGFmRNA、FGF、FGFR的表达明显减少；青年对照组和老龄对照组差异显著（P＜0．01）；老龄红景天素组FGFmRNA、FGF、FGFR明显高于老龄对照组（P＜0．05）。并且FGFR与FGF的表达呈明显的正相关。结论：大鼠嗅球组织的生长，发育与成纤维细胞生长因子及其受体信号传导密切相关；推测红景天素主要通过诱导大鼠嗅球FGF的表达，上调其受体FGFR的信号传导途径，发挥其对嗅球的抗衰老作用。     

红霉素干预对离体人鼻息肉嗜酸粒细胞凋亡基因表达的影响

目的　本实验探讨人鼻息肉组织中嗜酸粒细胞凋亡基因的表达以及红霉素干预对其表达的影响。方法　采用免疫组化方法检测 4 0例离体人鼻息肉组织中嗜酸粒细胞凋亡基因的表达 ,及15例经红霉素干预后 (外植块培养 )的离体人鼻息肉组织中嗜酸粒细胞凋亡基因表达的变化。结果　鼻息肉组织Bcl 2表达 5 % (2 / 4 0 ) ,Bax表达 6 5 % (2 6 / 4 0 ) ,Bcl xL 表达 4 0 % (16 / 4 0 ) ,Fas表达90 % (36 / 4 0 )。与下鼻甲黏膜相比 ,Bax和Fas表达差异有显著性 (P <0 0 5 )。经红霉素干预后 ,鼻息肉组织Bax表达明显上调 (93 3% ,14 / 15 ) ,与单纯用RPMI16 4 0培养者比较差异有显著性 (P <0 0 5 ) ,但Bcl xL和Fas表达差异无显著性。结论　Bax可能是鼻息肉组织嗜酸粒细胞凋亡抑制机制中最有意义的调控基因 ,红霉素可增强Bax的表达 ,是促进嗜酸粒细胞凋亡因素之一。     

变应性鼻炎大鼠鼻黏膜Bcl-2 mRNA及其蛋白的表达

目的 :探讨变应性鼻炎 (AR)大鼠鼻黏膜凋亡相关基因Bcl 2mRNA及其蛋白的表达 ,以进一步了解变应性鼻炎的发病机制。方法 :将 2 0只大鼠随机分为实验组和对照组 ,每组 10只。实验组以卵清蛋白腹腔注射致敏 ,继之鼻局部激发建立变应性鼻炎动物模型。取实验组与对照组动物鼻呼吸区黏膜 ,行Bcl 2mRNA原位杂交染色、Bcl 2与Bax蛋白免疫组化染色 ,并行苏木精 伊红染色以资对比。结果 :实验组Bcl 2mRNA与Bcl 2蛋白主要表达于鼻黏膜腺体细胞 ,其次是上皮层 ,鼻分泌物中可见大量呈强阳性反应的嗜酸性粒细胞 ,Bcl 2mRNA与Bcl 2蛋白表达水平显著高于对照组 (P <0 .0 5 ) ,Bax蛋白与Bcl 2蛋白表达部位一致 ,两组表达水平差异无统计学意义 (P >0 .0 5 )。结论 :变应性鼻炎大鼠鼻黏膜凋亡调控基因Bcl 2mRNA及其蛋白表达上调 ,是变应性鼻炎鼻分泌物增多及鼻黏膜上皮破坏的机制之一     

增殖和凋亡基因在喉上皮良恶性病变中的检测及其意义

目的 :探讨增殖相关基因Ki 6 7和凋亡相关基因Bax、Bcl 2在喉上皮良、恶性疾病中的表达和临床意义。方法 :采用免疫组织化学SP法检测存档石蜡标本喉癌、不典型增生、喉乳头状瘤、声带息肉、喉正常粘膜组织中Ki 6 7和Bax、Bcl 2表达。结果 :Ki 6 7在以上组织中的表达率依次为 5 2 .0 %、5 0 .0 %、4 7.8%、4 1.7%、10 .0 % ,喉癌明显高于喉正常粘膜 4 4 .0 % (P <0 .0 5 ) ,不典型增生明显高于喉正常粘膜 (P <0 .0 5 )。Bcl 2的表达率在不典型增生为 11.0 % ,在喉癌中为 4 4 .0 % ,其余 3组为 0 %。喉癌表达明显高于不典型增生和正常粘膜(P <0 .0 5 ) ,极显著高于喉乳头状瘤和声带息肉 (均P <0 .0 1)。Bax在各组织中表达率依次为 76 .0 %、77.8%、6 9.6 %、5 8.3%、80 .0 % ,各组差异无显著性意义。在喉乳头状瘤中Ki 6 7、Bcl 2表达成正相关 (P <0 .0 5 )。结论 :Ki 6 7、Bcl 2在喉乳头状瘤中的协同表达提示细胞内无法控制的增殖 ;二者有可能是癌变的前提 ,在癌前病变中有一定的诊断价值     

大鼠嗅球中凋亡相关基因蛋白和成纤维细胞因子表达的相 …

OBJECTIVE: To study the expression and distribution of apoptotic gene Bcl-2 and Bax on olfactory of rat, and its relation with the FGF and to investigate the mechanism of senile dysosmia. METHODS: Ten young (3 months) and ten senile rats (24 months) were used in this study. After the removal of the heads of these rats, olfactory bulb were immediately fixed with neutralized formalin, followed with paraffin-embedding, serial sectioning, immunohistochemiscal staining and microscopic observing. RESULTS: The positive expression rate of Bcl-2 and FGF on olfactory bulb in young rats was significantly stronger than that in senile rats (P < 0.01). The positive expression rate of Bax in senile rats was a little stronger than that in young rats, but the differences were not statistically significant. (P > 0.05) CONCLUSION: The expression of FGF and anti-apoptotic gene Bcl-2 on olfactory bulb decrease with the increase of age. This suggests neuron apoptosis on olfactory bulb are regulated by neurotrophic factor, and directly related with senile dysosmia. The mitral cell apoptosis as a result of FGF decreased on olfactory bulb may play a key role in the senile dysosmia.     

红景天素对老龄大鼠嗅球中成纤维细胞生长因子表达的影响及意义

目的　探讨红景天素对老龄大鼠嗅球中成纤维细胞生长因子（ｆｉｂｒｏｂｌａｓｔｇｒｏｗｔｈ ｆａｃｔｏｒ,ＦＧＦ）表达的影响及意义。方法　将２０ 只老龄大鼠随机分成注射红景天素实验组（１０ 只） 和注射等量生理盐水对照组（１０ 只）。分别取嗅球和嗅粘膜,经石蜡包埋,连续切片,免疫组化染色,光镜观察。结果　实验组嗅球中ＦＧＦ阳性表达率明显高于对照组（ Ｐ＜ ０．０１） ,而２ 组嗅粘膜均未见ＦＧＦ表达。结论　红景天素能够提高老龄大鼠嗅球中ＦＧＦ的阳性表达率,提示其对嗅系统可能具有抗衰老作用。     

红景天素对老龄大鼠嗅球中成纤维细胞生长因子表达的影 …

目的 探讨红景天素对老龄大鼠嗅球中成纤维细胞生长因子（ＦＧＦ）表达的影响及意义。方法 将２０只老龄大鼠随机分成注射红景天素实验组（１０只）和注射等量生理盐水对照组（１０只）。分别取嗅球和嗅粘膜,经石蜡包埋,连续切片,免疫组化染色,光镜观察。结果 实验组嗅球中ＦＧＦ阳性表达率明显高于对照组（Ｐ〈０．０１）,而２组溴粘膜均未见ＦＧＦ表达。结论 红景天素能够提高老龄大鼠嗅球中ＦＧＦ的阳性表达率,提示其对     

Apoptosis in the Aging Olfactory Epithelium

Objectives Olfactory receptor neurons undergo apoptosis at a baseline rate, probably secondary to environmental damage even in the absence of gross disease. The study demonstrates age‐related changes in expression of genes known to regulate apoptosis in the rat olfactory mucosa. These results are compared with gene expression in young rats and rats that have undergone surgical deafferentation of the olfactory receptor neurons. Study Design The olfactory mucosae from three groups of rats were studied: young, normal rats (age, 12 wk); old, normal rats (age, 24 mo); and young rats 9 days after bilateral removal of the olfactory bulb. Bulbectomy is known to produce an initial wave of apoptotic cell death in the population of olfactory neurons. At 9 days after the injury, the olfactory mucosae consist of an enhanced population of regenerating neurons destined to also undergo apoptosis, since their synaptic target (bulb) has been removed. Methods Ribonuclease protection assays and histological analysis of the three groups were performed. Results Ribonuclease protection assay analysis indicates that age induces increases in the expression of pro‐apoptotic genes in the olfactory mucosae similar to the increase seen after deafferentation (bulbectomy). Specifically, the expression of procaspase‐3 and bax was increased in aged animals and bulbectomized animals when compared with young, normal animals. Conclusions Aging induces changes in gene expression in the olfactory mucosae that appear to favor apoptosis, probably associated with increased fragility of olfactory receptor neurons in older animals. These changes may, at least in part, explain the age‐related decline in olfactory sensation and loss of olfactory receptor neurons seen in elderly patients.     

凋亡相关蛋白Bcl-2和Bax在慢性鼻及鼻窦炎伴嗅觉障碍患者嗅黏膜中的表达

目的研究Bcl-2和Bax在慢性鼻及鼻窦炎（CRS）伴嗅觉障碍患者嗅黏膜中的表达,探讨其对嗅觉神经元（olfactory receptor neurons,ORNs）凋亡的调节作用。方法采用康涅狄格化学感受临床研究中心所采用的嗅觉检查法——CCCRC（Connecticut ChemosensoryClinical Research Center）对46例行功能性鼻内镜手术的患者进行嗅觉评分并分组：A组,CRS伴嗅觉障碍25例;B组,CRS不伴嗅觉障碍10例;C组,单纯鼻中隔偏曲行鼻中隔矫正术11例。免疫组织化学方法检测Bcl-2、Bax在3组患者嗅黏膜中的表达。结果在ORNs中,A组Bcl-2和Bax表达显著高于B组（q=3.24、4.29,P均〈0.05）和C组（q=8.56、12.99,P均〈0.01）,A组Bcl-2/Bax比值显著低于B组（q=3.76,P〈0.05）和C组（q=6.67,P〈0.01）;在基底细胞中,Bcl-2在3组表达无显著差异（q=0.68、0.69、1.06,P均〉0.05）,Bax在A组的表达显著高于B组和C组（q=9.54、11.98,P均〈0.01）,A组Bcl-2/Bax比值显著低于B组和C组（q=5.48、9.14,P均〈0.01）;在A、B、C 3组ORNs和基底细胞中,Bcl-2/Bax比值与嗅觉评分均呈正相关（rA分别为0.5631、0.8926,rB分别为0.5700、0.7991、rC分别为0.5694、0.8121,P均〈0.01）。结论细胞凋亡参与了CRS伴嗅觉障碍患者ORNs的减少,Bcl-2和Bax在此过程中起了重要的调控作用,Bcl-2/Bax比值决定细胞是否凋亡。     

Age-related olfactory dysfunction: cellular and molecular characterization in the rat

BACKGROUND: Olfactory receptor neurons (ORNs) undergo apoptosis at a baseline rate even in the absence of obvious disease. Although the precise triggers of the apoptotic cascade are unclear, ORNs are exposed directly to the external environment, making them susceptible to injury. As an adaptive mechanism, mammals have the ability to replace lost ORNs throughout adult life from neuronal precursors within the olfactory epithelium (OE). In humans, this process fails with age as the surface area of the OE and the number of ORNs decline, coupled with a loss of clinical olfactory function. The question addressed in this study is whether this age-related failure of olfactory sensation is a result of a decrease in neuronal proliferation or an increase in ORN cell death. METHODS: To begin to address this question the ribonuclease protection assay was used to assess expression of apoptosis-related genes in rat OE as a function of age. Second, the terminal deoxynucleotide transferase end labeling assay was used to assess the percentage of ORNs undergoing apoptosis (apoptotic index) in three groups of animals: young (12 weeks), old (32 months), and bulbectomized rats. Bulbectomy is a standard model for ORN injury associated with a massive increase in ORN apoptosis and serves as a positive control. RESULTS: Ribonuclease protection assay data indicate an age-related increase in Bax, Bcl-xL, and procaspase-3 messenger RNA expression in aged compared with young rats. A similar but more pronounced increase in expression of these apoptotic-related genes is seen after bulbectomy. The terminal deoxynucleotide transferase end labeling assay also showed a statistically significant increase in the apoptotic index with both age and bulbectomy. CONCLUSION: Taken together, the current results indicate that aging and injury induce parallel changes in OE. Furthermore, these findings support the hypothesis that age-related olfactory dysfunction is, at least in part, related to an increase in ORN cell death.     

成纤维生长因子 2在慢性鼻窦炎嗅觉障碍嗅上皮中的表达

目的：探讨慢性鼻-鼻窦炎嗅觉功能障碍的发病机制和成纤维生长因子 2 (fibroblast growth factors 2, FGF2)在嗅上皮表达中的意义。方法：应用免疫组织化学技术检测32例慢性鼻窦炎患者和11例对照嗅黏膜FGF2的表达。结果：和对照组相比，慢性鼻窦炎患者嗅粘膜FGF2的表达明显减少（P＜0.05）。结论：慢性鼻-鼻窦炎嗅粘膜中促进再生因子FGF2明显减少。     

抑凋亡基因Bcl—X在鼻息肉中表达的初步研究

目的：探讨细胞凋亡基因Ｂｃｌ－２，Ｂｃｌ－Ｘ，Ｂａｋ和Ｂａｘ蛋白在鼻息肉中的表达及意义。方法：应用免疫组织化学方法（ＡＰＡＡＰ法）检测Ｂｃｌ－２，Ｂｃｌ－２，Ｂａｋ和Ｂａｘ蛋白在６例鼻息肉中的表达。结果：６例鼻息肉上皮Ｂｃｌ－Ｘ均呈强阳性表达，３例（３／６）Ｂｃｌ－２阳性表达，２例（２／６）Ｂａｋ弱阳性表达，１例（１／６）Ｂａｘ阳性表达。结论：鼻息肉上皮抑凋亡基因Ｂｃｌ－Ｘ蛋白表达上调和促调亡基因