【摘要】： 目的：研究兩種不同劑量的鏈霉素損傷后,小鼠前庭感覺上皮中毛細胞再生情況。 方法：通過左耳后半規(guī)管將兩種不同劑量的鏈霉素注入小鼠前庭,分別建立小鼠前庭感覺上皮中度和重度損傷模型。術(shù)后不同時間點行橢圓囊免疫熒光染色、掃描電鏡和樹脂切片,觀察不同程度損傷后橢圓囊感覺上皮的形態(tài)特征及毛細胞再生情況。 結(jié)果：橢圓囊前庭感覺上皮中度損傷后,毛細胞嚴重缺失,支持細胞保存良好；損傷后2周內(nèi)出現(xiàn)以幼小纖毛為特征的新生毛細胞,并隨著時間延長而增多；但是新生毛細胞的形態(tài)有別于正常毛細胞,而且再生的毛細胞數(shù)目有限。重度損傷后,毛細胞和支持細胞均受到嚴重損害,前庭感覺上皮被一極薄的單層扁平上皮取代；毛細胞的自我再生能力較中度損傷時極度減弱；部分細胞無毛細胞或支持細胞的特征。 結(jié)論：鏈霉素損傷后,小鼠前庭感覺上皮具有有限的自我再生能力。重度損傷極度降低了這種再生能力,前庭感覺上皮被一極薄的單層扁平上皮取代。 目的：探討小鼠前庭毛細胞再生過程中Notch信號途徑因子和Atoh1的作用。 方法：通過后半規(guī)管途徑將鏈霉素注入小鼠前庭,建立小鼠前庭感覺上皮中度損傷模型。然后我們分別用免疫熒光染色和實時定量PCR的方法檢測橢圓囊前庭感覺上皮中Notch信號途徑因子(Notch1, Jagged1, Hes1, Hes5)及Atoh1的蛋白和mRNA的表達。 結(jié)果：免疫組化結(jié)果顯示Jagged1表達于正常和損傷后支持細胞的細胞膜；Atoh1在正常小鼠前庭感覺上皮中無表達,但是毛細胞損傷后Atoh1表達增高,并可伴有myosinⅦa的共表達。實時定量PCR結(jié)果顯示小鼠前庭毛細胞再生過程中Hes5下降,Atohl上升,Notch1, Jagged1和Hes1無變化。 結(jié)論：Notch信號通路和Atoh1參與了小鼠前庭毛細胞損傷后的自我再生過程。再生的啟動時間可能早于過去的觀點,Atoh1的升高可能是多條信號通路共同作用的結(jié)果。 目的：研究兩種不同劑量的鏈霉素損傷后,小鼠前庭感覺上皮中神經(jīng)突觸、神經(jīng)纖維和前庭神經(jīng)節(jié)神經(jīng)元的變化。 方法：通過后半規(guī)管將兩種不同劑量的鏈霉素注入小鼠前庭,分別建立小鼠前庭感覺上皮中度和重度損傷模型。神經(jīng)絲蛋白200抗體免疫熒光染色觀察損傷后不同時間點橢圓囊前庭感覺上皮中神經(jīng)突觸和神經(jīng)纖維的變化；上前庭神經(jīng)節(jié)神經(jīng)元樹脂切片及定量分析,評價神經(jīng)元的存活情況。 結(jié)果：中度損傷和重度損傷后1月,神經(jīng)盞損傷嚴重,但是神經(jīng)纖維存活,上前庭神經(jīng)節(jié)神經(jīng)元形態(tài)正常,密度無明顯改變；重度損傷后3月,扁平上皮中無神經(jīng)盞,上前庭神經(jīng)節(jié)神經(jīng)元的密度明顯下降。 結(jié)論：小鼠前庭感覺上皮損傷后,神經(jīng)支配退化緩慢。這為干細胞移植或人工前庭植入研究提供了有利條件。
[Abstract]:Aim: to study the regeneration of hair cells in vestibular sensory epithelium of mice after two different doses of streptomycin. Methods: two different doses of streptomycin were injected into the vestibular of mice through the posterior semicircular canal of the left ear to establish the model of moderate and severe vestibular sensory epithelium injury. Immunofluorescence staining, scanning electron microscopy and resin sections were performed at different time points after operation to observe the morphological characteristics of sensory epithelium and hair cell regeneration after different degrees of injury. Results: after moderate injury of sensory epithelium of ovale vesicles, hair cells were seriously absent and Sertoli cells were well preserved, and new hair cells with the characteristics of young cilia appeared within 2 weeks after injury, and increased with the prolongation of time. However, the morphology of new hair cells is different from that of normal hair cells, and the number of regenerated hair cells is limited. After severe injury, both hair cells and Sertoli cells were seriously damaged, the vestibular sensory epithelium was replaced by a very thin monolayer flat epithelium, and the ability of self-regeneration of hair cells was extremely weakened when the injury was moderate. Characteristics of hairless cells or Sertoli cells in some cells. Conclusion: the vestibular sensory epithelium of mice has limited ability of self-regeneration after streptomycin injury. Severe injury greatly reduces this ability to regenerate and the vestibular sensory epithelium is replaced by a very thin monolayer flattened epithelium. Aim: to investigate the role of Notch signal pathway factor and Atoh1 in the regeneration of mouse vestibular hair cells. Methods: streptomycin was injected into the vestibule of mice through the posterior semicircular canal pathway to establish the model of moderate vestibular sensory epithelial injury in mice. Then immunofluorescence staining and real-time quantitative PCR were used to detect the expression of Notch signal pathway factor (Notch1, Jagged1, Hes1, Hes5), Atoh1 protein and mRNA in the vestibular sensory epithelium of elliptic sac. Results: the results of immunohistochemistry showed that Jagged1 was expressed in the membrane of Sertoli cells after injury and normal mice, but there was no expression of Atoh1 in vestibular sensory epithelium of normal mice, but the expression of Atoh1 was increased after hair cell injury with co-expression of myosin 鈪，

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