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关性及其意义

目的　明确成纤维细胞因子 (fibroblastgrowthfactor,FGF)和凋亡相关基因Bcl 2及Bax在大鼠嗅球中的表达及相关性 ,并探讨老化性嗅觉障碍的机制。方法　取健康大鼠青龄组 ( 3个月 ) ,老龄组 ( 2 4个月 )各 10只 ,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。结果青龄组大鼠嗅球中Bcl 2和FGF阳性细胞表达率分别为 ( 86 40± 5 2 2 ) %和 ( 70 2 5± 6 0 1) % ,显著高于老龄组的 ( 30 0 0± 4 12 ) %和 ( 2 4 10± 4 0 0 ) % ,二者呈明显正相关 ( x±s,P <0 .0 1,r =0 .8971) ;Bax阳性细胞表达率老龄组为 ( 98.2 4± 6 .5 0 ) % ,虽高于青龄组的 ( 82 .2 5± 6 .37) % ,但差异不显著。结论　随着年龄的增长嗅球中FGF表达减少 ,抗凋亡基因Bcl 2明显减少 ,提示嗅球中抗凋亡基因Bcl 2表达受成纤维细胞生长因子的调控 ,且与老化密切相关。     

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。     

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。     

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。     

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。     

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。     

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     

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。     

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.     

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。     

Protein kinase C in eosinophil proliferation and infiltration in nasal polyp tissue]

OBJECTIVE: To explore the significance of protein kinase C (PKC) in eosinophil (EOS) proliferation and infiltration in nasal polyp tissues. METHODS: With the methods of in situ hybridization and immunohistochemistry staining, PKC and apoptosis gene (Bcl-2, Bax) expression were measured between nasal polyps from 26 patients and inferior turbinate mucous membrane tissue (ITMMT) from 20 normal persons. EOS was identified with May-Grünwald-Giemsa (MGG) staining. RESULTS: In EOS, the positive cell expressive rate of Bcl-2 mRNA and its protein were significantly higher in group of nasal polyps than that in group of ITMMT (P < 0.01). Although the positive cell expressive rate of Bax mRNA and its protein were slightly higher in group of nasal polyp tissue than that in group of ITMMT, the difference was not significant (P > 0.05). PKC was expressed in EOS of all 26 cases from nasal polyp tissues, but in 20 cases from ITMMT, only 7 cases showed occasional PKC expression. In both groups, PKC positive cell expression was significantly different, and the expression of PKC and Bcl-2 mRNA as well as their protein in EOS of nasal polyps showed remarkably positive relation (r1 = 0.0875, r2 = 0.0823, P < 0.01). CONCLUSION: PKC expression increasing in EOS of nasal polyp tissues was closely associated with apoptosis inhibiting. So we presumed that EOS inhibiting in nasal polyp tissues was obtained by activating PKC signal transduction pathway.     

阻塞性睡眠呼吸暂停低通气综合征上气道开大肌凋亡现象研究

目的:观察阻塞性睡眠呼吸暂停低通气综合征(OSAHS)患者腭帆张肌中Bax、Bcl-2的表达水平,探讨OSAHS患者上气道开大肌肌纤维是否存在凋亡现象。方法:选取30例OSAHS患者作为实验组,10例已排除OSAHS的慢性扁桃体炎患者作为对照组。采用免疫组织化学Elivison二步法和计算机图像分析系统检测两组腭帆张肌中Bax、Bcl-2的表达水平并进行对比分析。结果:①实验组Bax的表达水平和Bax/Bcl-2的比值明显高于对照组,差异有统计学意义(P<0.05);②Bax的表达水平与呼吸紊乱指数呈正相关(r=0.697,P<0.01)。结论:OSAHS患者上气道开大肌存在凋亡现象,且病情越重凋亡的程度也越严重。     

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.     

鼻息肉中诱导型一氧化氮合酶、Bax及Bcl-2的表达及相关性研究

目的:检测鼻息肉组织中诱导型一氧化氮合酶(iNOS)和凋亡相关基因Bax、Bcl-2的表达及相关性。方法:30例鼻息肉组织,应用TUNEL方法检测其凋亡情况;免疫组织化学SABC法检测iNOS及Bax、Bcl-2的表达;western blot方法检测iNOS、Bax、Bcl-2的表达。结果:①TUNEL方法检测凋亡细胞位于鼻息肉表层上皮细胞及腺体上皮细胞,均为弱阳性。②免疫组织化学法检测iNOS、Bax、Bcl-2均表达于鼻息肉组织上皮细胞、腺上皮细胞、血管内皮细胞和浸润的炎性细胞,阳性表达定位于细胞质,其中Bax表达为弱阳性,iNOS、Bcl-2表达为强阳性。③Western blot检测3种蛋白均有表达条带,iNOS、Bal-2条带清晰,Bax条带软弱,差异有统计学意义(P<0.01);iNOS与Bcl-2的灰度值比值呈正相关(r=0.851,P<0.01);iNOS与Bax的灰度值比值呈负相关(r=-0.714,P<0.01)。结论:鼻息肉组织中存在抗凋亡和促凋亡2种蛋白,iNOS抑制细胞凋亡,可能对鼻息肉的发生、发展起重要的促进作用。