损伤性嗅觉诱发电位的实验研究

目的　建立损伤性嗅觉功能障碍动物模型 ,了解电刺激嗅觉诱发电位的特性及变化规律。方法　通过立体定位和电解损毁技术 ,对动物一侧或双侧嗅球造成损伤 ,动态观察损伤后不同时间点 (2 4h、4 8h和 1周 )嗅觉诱发电位的变化 ,并观察嗅球与嗅黏膜病理和超微结构改变。结果一侧嗅球损伤后 ,诱发电位N2波消失 ,N1和P1波潜伏期延长 ,波幅减小 ;双侧嗅球损伤后 ,表现为N1、N2波消失 ,仅记录到P1波 ,其潜伏期明显延长 ,波幅变化较大 ,以上改变均以 2 4～ 4 8h变化最显著。组织病理学和超微结构显示 ,2 4～ 4 8h嗅球周围大量炎性细胞浸润 ,神经元变性样改变。结论嗅球损伤程度不同 ,对嗅觉诱发电位的影响也不相同 ,这种改变是以其组织病理和超微结构变化为基础的。     

嗅神经离断后大鼠嗅黏膜气味受体的表达变化

明显减少,差异具有统计学意义(t=63.960,P=0.000).随着术后恢复时间的延长,嗅神经离断侧再生嗅黏膜表达Olr1493基因受体的细胞逐渐增多,至第6周时基本和对照侧一致,分别为(417.8±32.4)个和(445.3±10.0)个,差异无统计学意义(t=2.600,P=0.060).结论 大鼠嗅神经离断后再生嗅感觉神经元及其受体数量可以恢复至正常水平,表达位置亦未发生变化.     

分鼻侧评估嗅觉功能及单侧嗅觉减退的临床分析

目的 了解嗅觉减退患者左右两侧嗅觉功能损伤程度是否相同.方法 对104例以嗅觉减退为主诉的患者,通过病史采集了解嗅觉减退的相关病史及其现况,耳鼻咽喉科常规检查及鼻窦CT检查排除鼻腔结构异常或仅发生于单侧鼻腔-鼻窦的病变,并分别对左、右鼻侧进行T&T嗅觉计嗅觉识别阈测试,评估患者每侧嗅觉功能.结果 本组104例嗅觉减退患者,90例(86.5％)患者双侧嗅觉功能水平相同,14例(13.5％)患者双侧嗅觉功能水平不同,其中6例患者表现为单侧(左侧或右侧)嗅觉功能正常,而另一侧嗅觉功能减退.14例左右鼻侧嗅觉功能不同的患者,其常规查体及影像学检查均未见明显鼻腔结构异常或发生于单侧鼻腔鼻窦的病变.结论 人类的左、右两个鼻腔分别拥有一个完整的嗅觉传导系统,尽管双侧受损概率相同,但受损程度可不同.单侧嗅觉减退作为嗅觉障碍的一种特殊表现形式,只有通过分鼻侧进行嗅觉功能评估,才能更全面地了解患者的嗅觉功能.     

The effect of basic fibroblast growth factor on the regeneration of guinea pig olfactory epithelium

Olfactory receptor cells are widely thought to regenerate after degeneration and also thought to show turnover in normal circumstances in animal olfactory epithelium. The identity of the factor that controls proliferation and differentiation of olfactory receptor cells is a very important problem that has yet to be resolved. In this study, the mitogenic effects of epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) on olfactory receptor cells in guinea pig olfactory epithelium was examined. The intraperitoneal injection of 1,000 ng bFGF/day for 14 days increased the cells in proliferation detected by immunostaining with proliferating cell nuclear antigen (PCNA), while neither EGF nor low-dose bFGF had any effect. These results support the idea that an adequate dose of bFGF plays an important role in the neurogenesis in the olfactory epithelium. Further study is needed to clarify the efficacy of bFGF in the damaged olfactory epithelium, but bFGF may provide a therapeutic option for olfactory disturbances caused by complete or partial loss of olfactory receptor cells. Received: 3 October 2001 / Accepted: 10 October 2001     

放射线损伤对小鼠嗅上皮细胞凋亡及再生的影响

目的研究放射线损伤后小鼠嗅上皮细胞凋亡和再生情况,探讨放射治疗后嗅觉障碍的致病机制。方法以4Gy的X射线照射小鼠鼻部,建立放射线损伤小鼠嗅黏膜的动物模型分别于照射前及照射后第1,3,6,12,60天分批处死动物取材;TUNEL法检测嗅上皮中细胞凋亡,BrdU掺入免疫组化检测基底细胞再生。结果放射线损伤后小鼠嗅上皮凋亡细胞显著增多（P〈0.01）,基底细胞再生也相应增多（P〈0.01）,但再生细胞少于凋亡细胞（P〈0.01）。结论放射线损伤可促进嗅上皮细胞凋亡及基底细胞再生,但再生细胞少于凋亡细胞,二者失衡可能是放射治疗后嗅觉障碍的致病机制之一。     

Correlation and agreement of olfactory perception assessed by the Connecticut Chemosensory Clinical Research Center olfactory test and the Brief-Smell Identification Test

IntroductionAssessing olfactory perception in olfactory disorders is of utmost importance in therapy management. However, the University of Pennsylvania Smell Identification Test and the Sniffin’ Sticks are the only tests validated in Brazil.ObjectivesTo evaluate the correlation and agreement between the Chemosensory Clinical Research Center olfactory test and the Brief-Smell Identification Test – University of Pennsylvania Smell Identification Test – in healthy participants and in participants with olfactory disorders based on the results and technical aspects of both tests.MethodsFifty participants without olfactory complaints and 50 participants with olfactory disorders who underwent the Chemosensory Clinical Research Center olfactory test and the Brief-Smell Identification Test were included. The following tests were used for statistical analysis: Mann-Whitney U test, Spearman’s correlation, intraclass correlation coefficient and Bland-Altman plot. An alpha error (significance level) of 0.05 was considered in the statistical analysis.ResultsBoth tests were effective in distinguishing the groups without the presence of overlapping values ??for the measured markers. Additionally, there was a strong correlation between Spearman’s correlation and intraclass correlation coefficient between the tests and for both nostrils. However, the correlations were lower when the groups were individually evaluated. The Bland-Altman plot showed no bias when all participants were simultaneously evaluated.ConclusionsThe tests to assess olfactory perception presented a high level of agreement. In our sample, we could infer that the Connecticut Chemosensory Clinical Research Center olfactory test is similar to the Brief-Smell Identification Test and can be used in the routine diagnosis of patients with complaints of olfactory disorders, considering the advantage of its low cost.     

嗅裂疾病三例

目的报道一种新的嗅觉障碍疾病——嗅裂疾病，总结其临床特征、影像学特点以及治疗尝试。方法本组3例，均以完全失嗅为主诉。详细采集病史，行耳鼻咽喉科常规体检、鼻内镜检查。主观嗅功能测试使用T＆T标准嗅觉计行嗅觉察觉阈和识别阈测试。以醋酸异戊酯为刺激剂行嗅觉事件相关电位测试，刺激量为2ml。行鼻窦冠状位和轴位CT扫描。治疗方案：①抗生素治疗半个月；②糖皮质激素治疗半个月。结果鼻内镜检查患者双侧嗅裂处黏膜肿胀，嗅裂消失。T＆T主观嗅觉测试均为完全失嗅，最大嗅刺激未引出嗅觉事件相关电位。鼻窦CT提示局限于双侧嗅裂的软组织影。抗生素治疗嗅觉无改善。局部和全身糖皮质激素治疗，1例嗅觉改善，但停药后嗅觉又消失，再次使用糖皮质激素治疗无效。另2例嗅觉无改善。结论嗅裂疾病是一种以嗅觉障碍为主诉，病变局限于嗅裂的疾病，鼻窦CT有助于确诊，抗生素和糖皮质激素治疗效果不理想。     

嗅觉诱发电位的实验研究

目的：观察电刺激家兔嗅区黏膜记录的诱发电位的波形及其稳定性，探讨一种可以客观评价嗅觉功能的方法。方法：将双极刺激电极经家兔前鼻孔置于嗅区黏膜，给予电刺激，在颅顶记录嗅觉诱发电位(OEPs)；改变刺激参数和部位，观察对电位潜伏期、阈值和波形的影响。结果：在颅顶前方近嗅球处记录到一组波形稳定的N1-P1-N23相复合波。潜伏期为：N1波20．6ms，P1波33．5ms，N2波58．3ms；改变刺激强度对各波无明显影响；改变刺激频率对各波影响较大，频率过高．波形分化较差。结论：正常OEPs是一组波形稳定的负-正-负3相复合波，波形稳定、重复性好，各参量可作为评价嗅觉功能变化的有用指标。     

Validation of the Connecticut olfactory test (CCCRC) adapted to Brazil

IntroductionOlfactory changes are quite common in the population, causing a significant impact on the quality of life. Documentation of the olfactory function is essential for the diagnosis, treatment and follow-up of patients with inflammatory diseases of the upper airways, neurodegenerative diseases or viral infections. Among the different existing smell tests, the CCCRC is an inexpensive test, easy to apply, but it has not yet been evaluated on a large scale in the Brazilian population.ObjectiveTo validate the CCCRC smell test, after adaptation for the Brazilian population, evaluating the performance of healthy volunteers and the stability of the test in retests.MethodsIn this study, we carried out a cultural adaptation of the CCCRC test to Brazil. To validate and determine the normality scores, we applied the test to 334 healthy volunteers, aged >18 years of age. The retest was also carried out in up to four weeks on 34 additional volunteers to assess validity of the results.ResultsWhen evaluating the participants’ performance, normosmia and mild hyposmia values were obtained in more than 95% of them. Women (58.4%) showed better accuracy than men (41.6%): p < 0.02, and individuals over 60 years of age showed worse performance (median: 6; 75^th percentile: 6.5; 25^th percentile). The test and retest of the 34 volunteers demonstrated that there was agreement (ICC, intraclass correlation coefficient) considered good in the left nostril (ICC = 0.65) and excellent in the right nostril (ICC = 0.77) in the combined score.ConclusionThe CCCRC test adapted to Brazil showed normal values, similar to the originally-described test and validations in other countries, with a high reproducibility rate. Considering the highly favorable cost-benefit ratio, the adapted CCCRC is a very useful tool for measuring olfactory function in the Brazilian population.     

上呼吸道感染后嗅觉障碍患者嗅上皮超微结构观察

目的通过对上呼吸道感染后嗅觉障碍的患者鼻腔大体及嗅上皮超微结构的研究，从形态学上观察嗅觉减退或丧失的超微结构改变。方法选择上呼吸道感染后嗅觉减退或丧失患者10例，用T＆T嗅觉测试法测试患者的嗅觉功能。常规前鼻镜、鼻内镜下对鼻腔大体结构进行观察，鼻内镜下钳取嗅区黏膜行透射电镜超微结构观察。结果上呼吸道感染后嗅觉障碍患者嗅黏膜超微结构有以下变化：①嗅上皮结构层次仍能保持，但细胞间隙增宽；②上皮表面嗅泡明显减少，即使嗅泡存在，其末端的纤毛也明显减少，部分嗅泡呈空泡状改变；③微绒毛细胞和支持细胞表面的微绒毛减少或缺失；④支持细胞的细胞核变形或固缩，嗅细胞的树突水肿变形，细胞器减少。结论上呼吸道感染后嗅觉功能障碍与嗅黏膜上皮超微结构的改变密切相关。患者嗅泡及嗅泡内纤毛缺失，微绒毛细胞及支持细胞的微绒毛减少是引起嗅觉减退的主要原因，支持细胞胞核的变形及嗅细胞树突的形态学改变与嗅觉改变相关。     

Possible effects of low testosterone levels on olfactory function in males

IntroductionFunctions attributed to androgens have increased, ranging from the role in hypothalamic–pituitary–gonadal axis and reproductive behaviors to modulation of cognition, mood and some other functions. Sex differences and changes in circulating sex hormones affect human sensory function. In the literature, authors reported this kind of influence for olfaction predominantly in females.ObjectiveTo investigate the effects of low testosterone levels on olfactory functions in males, in this prospective clinical study.MethodsMale patients diagnosed with prostate cancer were included. Thirty-nine patients with prostate cancer whose testosterone levels were lower than 50 ng/dL due to castration, were the study group. Thirty-one patients with prostate cancer who were not castrated with testosterone levels higher than 50 ng/dL were selected as the control group. Acoustic rhinometry and peak nasal inspiratory flow tests were performed for all participants; and for evaluation of olfactory function, both groups completed the Connecticut chemosensory clinical research center olfactory test.ResultsThe mean ages of the patients and controls were 69.6 ± 7.2 (57–89) and 66.3 ± 5.8 (50–78) years, respectively (p = 0.039). There was a significant difference between groups in terms of testosterone levels (p < 0.0001). The multivariate logistic regression revealed testosterone level as the only predictive factor determining the difference between the groups. In terms of olfactory parameters, all scores were lower in the emasculated group (butanol threshold test p = 0.019, identification p = 0.059, and Connecticut center score p = 0.029) There was a significant correlation between testosterone levels and olfactory parameters (p = 0.023; p = 0.025 for identification and Connecticut center scores, respectively).ConclusionLow testosterone levels in males have negative effects on olfactory functions. Further molecular research is required to understand the connection between testosterone and olfaction.     

Olfactory rehabilitation and olfactory bulb volume changes in patients after total laryngectomy: a prospective randomized study

IntroductionAfter total laryngectomy, decreased olfactory function and olfactory bulb volume shrinkage have been reported to occur due to olfactory deprivation caused by nasal airflow interruption. There is evidence that the olfactory system can be modulated by repeated exposure to odors in a procedure called olfactory training. However, it is not known whether any recovery of the lost olfactory bulb volume is possible by eliminating olfactory deprivation via olfactory rehabilitation long after laryngectomy.ObjectiveThis study examined the recovery of olfactory function and the change in olfactory bulb volume via long-term olfactory rehabilitation after total laryngectomy.MethodsPossible causes of olfactory dysfunction in the study participants were evaluated by collecting detailed anamnesis. As olfactory tests, orthonasal butanol threshold and odor discrimination tests were performed. Three-dimensional olfactory bulb volumes were calculated using manual segmentation on T2-weighted coronal magnetic resonance images. In olfactory rehabilitation, four different odors were applied to all patients orthonasally, using a larynx bypass technique for 30 min per day for 6 months. Olfactory tests were performed before the rehabilitation and after 6 months of rehabilitation, and olfactory bulb volume measurements were performed using magnetic resonance images.ResultsEleven patients diagnosed with advanced laryngeal cancer who underwent total laryngectomy and postoperative radiotherapy with a follow-up of 5–10 years were included in the study. All patients were male, and the mean age was 58.18 ± 4.17 years. In total laryngectomized patients, the olfactory bulb volumes measured on magnetic resonance images were 42.25 ± 12.8 mm³ before and 55.5 ± 11.22 mm³ after rehabilitation, and this increase was highly significant. Olfactory test scores were 2.3 ± 1.27 before and 4.39 ± 0.86 after rehabilitation, and this increase was also highly significant.ConclusionAs a result of the olfactory rehabilitation applied by providing orthonasal air flow, the olfactory function lost after total laryngectomy was improved considerably, and the olfactory bulb volume was significantly increased. The increase in olfactory bulb volume in total laryngectomy patients via olfactory rehabilitation to eliminate olfactory deprivation due to nasal airflow interruption was demonstrated for the first time in this prospective longitudinal study.     

Examination and classification of human olfactory mucosa in patients with clinical olfactory disturbances

Summary To determine objectively the degree of olfactory disturbance, we biopsied the olfactory mucosa from patients who complained of anosmia. The olfactory disturbances in this study were caused by choanal atresia, chronic sinusitis, viral inflammation, and head trauma, as well as by congenital and idiopathic anosmia. The biopsy specimens were examined by light microscopy and the degree of mucosal degeneration present was classified according to five grades. The clinical courses of the patients studied paralleled the changes found in the olfactory mucosa.     

外伤后失嗅患者的嗅觉事件相关电位和MRI评估

目的 探讨嗅觉事件相关电位(olfactory event related potentials,OERP)和嗅觉通路MRI对外伤后失嗅的评估价值.方法 回顾性分析24例外伤后失嗅患者的临床资料.所有患者均行详细的病史采集、全面体检、T&T嗅觉检查、鼻内镜检查、OERP测试、颅脑CT和嗅觉通路MRI检查.结果 主观嗅觉测试:双侧完全失嗅20例;单侧完全失嗅,对侧嗅觉减退2例;单侧完全失嗅,对侧嗅觉正常2例.OERP测试:双侧最大嗅刺激均不能引出OERP者20例,单侧最大嗅刺激不能引出OERP者4例;单侧能引出OERP者4例,其中2例能正常引出,另2例OERP幅值下降且潜伏期延长.氨气刺激均能引出鼻内三叉神经化学感受事件相关电位.嗅路MRI检查:嗅球损伤24例次(100%),额叶直回损伤22例次(91.7%),额叶眶回损伤16例次(67%),远端嗅束和颞叶损伤各2例次(8%).结论 OERP能对外伤后嗅觉进行定性和定量的客观整体评估;嗅路MRI能对外伤后失嗅的损伤部位、程度进行客观、精确的评估.两者结合能对嗅觉功能进行全面、客观评价.     

Age-related changes in dividing cells of the olfactory epithelium of the maturing guinea pig

Changes in dividing cells of the olfactory epithelium from guinea pigs of different ages were examined by immunohistochemical staining using anti-proliferating cell nuclear antigen antibody. Numerous dividing cells were scattered diffusely in the basal layer of the olfactory epithelium at 1 and 2 months following birth and then gradually decreased with maturation until 4 months. Findings then remained constant between 4 and 24 months. Subsequently, cell numbers were found to decrease as animals became older. The number of olfactory receptor cells did not vary significantly between 1 and 30 months. Although no correlation could be found between the numbers of dividing cells and olfactory receptor cells, it is still possible that the longevity of the olfactory receptor cells changes to maintain the overall size of the neuronal population. Received: 18 February 1998 / Accepted: 9 April 1998     

Olfactory mucosa histological findings in laryngectomees

After total laryngectomy, the patients often report immediate and marked olfactory deficit. The aim of this study was to determine whether hyposmia in laryngectomees reflects olfactory epithelial damage. Ten laryngectomized patients and ten rhinologically normal subjects were subjected to olfactory testing, after which histological examination of biopsied olfactory mucosa was performed. Olfactory testing in laryngectomees revealed a marked reduction in odor perception. Histological examination of olfactory mucosa specimens showed that in laryngectomees some neuroepithelial structural features were comparable with those found in normal subjects. However, additional signs of damage were also observed, consisting mainly of various degrees of epithelial degeneration, above and beyond those that are characteristic of physiological epithelium turnover. These different degenerative features consisted of severe damage to the neuroepithelium, culminating in complete topical loss. Bowman's glands were also observed to be involved in the degenerative process. Laryngectomy-induced hyposmia seems to be correlated with the almost complete loss of nasal airflow due to the disconnection between the upper and lower airways, which prevents odor molecules from reaching the olfactory area, together with degenerative phenomena, which affect the neuroepithelium, and consequent failure in neurosensorial performance.     

神经特异性烯醇化酶及嗅标记蛋白在不同胎龄胎儿嗅黏膜中的表达

目的　探讨神经特异性烯醇化酶 (neuron specificenolase ,NSE)及嗅标记蛋白 (olfactorymarkerprotein ,OMP)在不同胎龄胎儿嗅黏膜中的表达。方法　以免疫组化方法检测NSE及OMP在1 2、1 6、2 0、2 4、2 8和 34周 6例不同胎龄胎儿嗅黏膜中的表达。结果　NSE免疫阳性反应在孕 1 2～ 34周的胎儿嗅黏膜中均有表达 ,各胎龄胎儿嗅黏膜切片中均可见大量阳性着色的双极嗅神经细胞。孕1 2周时 ,阳性细胞数量很多 ,排列紧密 ,胞体多位于嗅上皮中下部且阳性嗅神经上皮占据胎儿鼻腔上2 /3的黏膜 ,随着孕龄的增大 ,阳性细胞逐渐成多层次排列 ,所占鼻腔黏膜面积却逐渐减小 ,至孕 34周时 ,仅局限于鼻腔上 1 /3黏膜。OMP免疫反应在孕 1 2周胎儿鼻腔上 2 /3黏膜切片中仅发现少量阳性着色的双极神经细胞 ,明显少于同龄胎儿NSE阳性反应细胞。随着胎龄的增加 ,OMP阳性细胞逐渐增多 ,且胞体多位于嗅上皮中上部 ,但其数量仍相对少于同龄NSE阳性细胞。结论　人胚胎 1 2周时嗅黏膜中已有大量的嗅神经细胞 ,其中少数嗅神经细胞已开始发育成熟。随着胎龄的增大 ,嗅上皮面积逐渐缩小 ,但成熟的嗅神经细胞逐渐增多 ,胚胎发育后期的胎儿嗅化学感受器发育已趋于成熟     

嗅神经切断对小鼠嗅感觉神经元的影响

目的分析嗅神经切断对小鼠嗅感觉神经元（olfactory receptor neurons，ORN）凋亡情况的影响，并探讨这一造模方法的可靠件。方法取2月龄C57小鼠33只，随机分组后，实验组小鼠行嗅神经切断术，通过辣根过氧化物酶（horseradish peroxidase，HRP）顺行神经示踪验证造模成功与否。以脱氧核苷酸转移酶介导的牛物素dUTP缺口末端标记技术（TdT mediated deoxyuridine triphosphate—biotin nick end labelling，TUNEI。）观察术后8h、2d、3d和5d嗅上皮中ORN的凋亡情况，同时在蛋白和mRNA水平观察成熟ORN的特异性标记蛋白——嗅标记蛋白（olfactory marker protein，OMP）在嗅上皮巾的表达情况。结果嗅神经切断术后嗅球中无HRP标记。TUNEL和OMP阳性反应发生于ORN，嗅神经切断术后TUNEL阳性细胞数显著增多，并于术后第2天达到高峰，与此同时OMPmRNA的表达水平开始显著下降，并住术后第5天降至更低，嗅上皮的厚度也相应变薄。结论本实验所采取的造模方法可以较可靠地切断小鼠的嗅神经，并造成小鼠嗅上皮中ORN的同步凋亡。     

Variables associated with olfactory disorders in adults: A U.S. population-based analysis

ObjectiveOlfactory dysfunction is known to have significant social, psychological, and safety implications. Despite increasingly recognized prevalence, potential risk factors for olfactory loss have been arbitrarily documented and knowledge is limited in scale. The aim of this study is to identify potential demographic and exposure variables correlating with olfactory dysfunction.MethodsCross-sectional analysis of the 2011–2012 and 2013–2014 editions of the National Health Examination and Nutrition Survey was performed. The utilized survey reports from a nationally representative sample of about 5000 persons each year located in counties across the United States. There is an interview and physical examination component which includes demographic, socioeconomic, dietary, and health-related questions as well as medical, dental, physiologic measurements, and laboratory tests. 3594 adult respondents from 2011 to 2012 and 3708 respondents from 2013 to 2014 were identified from the above population-based database. The frequency of self-reported disorders as well as performance on odor identification testing was determined in relation to demographic factors, occupational or environmental exposures, and urinary levels of environmental and industrial compounds.ResultsIn both subjective and objective analysis, smell disorders were significantly more common with increasing age. While the non-Hispanic Black and non-Hispanic Asian populations were less likely to report subjective olfactory loss, they, along with Hispanics, performed more poorly on odor identification than Caucasians. Those with limited education had a decreased prevalence of hyposmia. Women outperformed men on smell testing. Those reporting exposure to vapors were more likely to experience olfactory dysfunction, and urinary levels of manganese, 2-Thioxothiazolidine-4-carboxylic acid, and 2-Aminothiazoline-4-carboxylic acid were lower among respondents with subjective smell disturbance. In odor detection, elevated serum levels of lead and urinary levels of 2,4 dichlorophenol were associated with anosmia and hyposmia, respectively.ConclusionsThis study provides current, population-based data identifying demographic and exposure elements related to smell disturbances in U.S. adults. Age, race, gender, education, exposure to vapors, urinary levels of manganese, 2-Thioxothiazolidine-4-carboxylic acid, 2-Aminothiazoline-4-carboxylic acid, 2,4 dichlorophenol, and serum lead levels were all implicated in smell disturbance. Care should be taken in interpretation due to lack of consistency between subjective and objective measures of olfaction as well as limitations related to population-based data. Prospective trials are indicated to further elucidate these relationships.     

嗅感觉神经细胞的分离与培养

目的 建立哺乳动物嗅感觉神经细胞体外培养的方法。方法取成年家兔的嗅区粘膜,用胰酶和胶原酶消化法进行嗅感觉神经细胞的分离与原代培养,并应用抗神经微丝蛋白(NFP)抗体、抗嗅觉标记蛋白(OMP)抗体进行免疫细胞化学染色,同时采用甲苯胺蓝神经特异性染色和扫描电镜对其进行鉴定。结果 从嗅粘膜分离以及原代培养得到的嗅感觉神经元,光镜、电镜下为典型的双极神经元,OMP、NFP阳性,神经元特异性染色阳性。分离的嗅感觉神经元可在体外存活数小时,其细胞形态、生存数量和存活时间均能满足体外实验的基本要求。结论 本实验在家兔建立了较稳定、可靠的嗅感觉神经细胞分离和原代培养方法。为深入开展嗅感觉神经细胞的离体研究,提供了较为理想的材料来源和技术保障。