G蛋白偶联受体30在声带中表达的观察

发布时间：2018-06-13 15:36

本文选题：GPR30 + 声带　；参考：《复旦大学》2012年硕士论文

【摘要】：[目的]：育龄期女性体内性激素水平会随月经周期而呈现周期性波动,这种雌激素浓度的波动可以影响女性的嗓音,研究表明,约有1/3的育龄期妇女会经历经前期发声综合征,表现为发声易疲劳、音域降低、一些泛音丢失、以及声强减弱等；喉镜下可表现为声带粘膜水肿,血管充血及炎症。动物模型实验亦表明,E2缺乏同样可使大鼠喉部粘膜发生充血、水肿及炎症,与育龄期女性声带粘膜的周期性表现类似。E2对嗓音的这种影响机制,到目前为止还不清楚,多数研究认为声带上无雌激素受体(ER)表达,为数不多的有关声带中SHRs的文献样本样相对小,报道的ER表达的阳性率也低。GPR30是一种能够与E2特异性结合的G蛋白,介导某些快速、非基因效应,最近被认为是一种新的雌激素受体,GPR30可存在于很多组织器官中,近来发现,在一些病理过程中,GPR30可通过抑制炎症反应来解缓机体所受到的损伤。因此我们推测GPR30可能也存在于声带,并且参与了E2介导的炎症过程,从而使很多育龄期女性在体内激素生理性波动时,声带粘膜出现短暂的充血、水肿等炎症反应。为此,本实验旨在了解GPR30在人及大鼠声带组织中的表达及其可能的意义。 [方法]：获取大鼠声带标本,收集人良性病变声带标本及喉切除的喉癌病例中未受肿瘤组织侵犯的瘤周声带标本。采用Rt-PCR的方法以及免疫组织化学方法观察大鼠及人VF组织中GPR30的表达及分布。利用χ2检验的方法统计不同性别、年龄及不同声带病变中GPR30表达有无差异。结合近年来相关文献,分析GPR30在声带因E2波动而产生的炎症反应过程中可能发挥的作用。 [结果]： 1.GPR30mRNA在所有大鼠声带标本(n=12)中均有表达；在人声带组织中,有19/29例标本可见有GPR30mRNA的表达。 2.免疫组织化学结果表明,GPR30蛋白在34/63例中有表达。GPR30位于VF上皮,可表达于胞膜,胞质和胞核中。 3.GPR30在VF不同病变类型中的表达：(1)GPR30mRNA在瘤周声带中的表达率为55.6%(5/9例),在VF良性病变组织中GPR30mRNA的阳性率为70%(14/20例)。(2)免疫组化结果表明,在癌前病变声带白斑及瘤周声带组织中,GPR30蛋白的阳性表达率为60.9%(14/23例),在其他VF的良性病变中,GPR30蛋白的阳性表达率为50%(20/40例)。 4.GPR30的表达部位：(1)在VF良性病变中,GPR30主要表达于上皮细胞的胞膜上,少数表达于细胞核。(2)而在瘤周声带组织,GPR30主要表达于上皮,VF固有层及肌层细胞均未见有表达,上皮细胞中GPR30定位主要是位于胞核。 5.在VF良性病变标中,GPR30在mRNA水平及蛋白水平表达具有性别及年龄差异,但无统计学意义。 [结论]： 1.本研究通过Rt-PCR和免疫组化的方法,首次在大鼠及人声带中观察到GPR30的表达。 2.GPR30主要表达于VF上皮细胞,在固有层及肌层均未见表达；而且,GPR30的表达部位与所处环境有关,良性病变主要表达于胞膜,而白斑及瘤周声带组织GPR30主要位于胞核。 3.GPR30可能参与了女性在月经周期中因E2波动而导的声带炎症反应。
[Abstract]:Purpose :

The sex hormone level of female in the childbearing age can fluctuate with the menstrual cycle , and the fluctuation of estrogen concentration can affect the female voice . The study shows that about 1 / 3 of the women in the childbearing age can experience the premenstrual syndrome of phonation , which shows that the vocalization is easy to fatigue , the sound field is reduced , some overtones are lost , and the sound intensity is weakened , etc .
In this paper , it is not clear that the expression of estrogen receptor ( ER ) in the vocal cord is similar to that of the female vocal cord mucosa in the childbearing age .

The method comprises the following steps of :

The expression and distribution of GPR 30 in rat and human VF tissues were observed by means of the method of Rt - PCR and immunohistochemical method . The expression and distribution of GPR 30 in rat and human VF tissues were observed by the method of Rt - PCR and immunohistochemical method .

The results are as follows :

1 . The mRNA was expressed in all rat vocal cord specimens ( n = 12 ) .
In human vocal cord tissue , there were 19 / 29 specimens showing the expression of GPRS 30mRNA .

2 . The results of immunohistochemistry showed that the pr30 protein was expressed in 34 / 63 cases , which could be expressed in the membrane , cytoplasm and nucleus of VF epithelium .

( 2 ) The positive expression rate was 60.9 % ( 14 / 23 ) and 50 % ( 20 / 40 ) in the benign lesions of other VF .

4 . The expression site : ( 1 ) In benign lesions of VF , the majority of the cells were expressed in the membrane of epithelial cells , and a small number was expressed in the nucleus .

5 . In benign lesions of VF , there was no significant difference in mRNA level and protein level in the expression of mRNA and protein level , but there was no statistical significance .

Conclusion :

1 . In this study , the expression of GPRS 30 was first observed in rat and human vocal cords by the method of Rt - PCR and immunohistochemistry .

2 . There was no expression in lamina propria and myometrium , mainly expressed in VF epithelial cells .
Moreover , the expression site of GPRS 30 is related to the environment in which benign lesions are mainly expressed in the cell membrane , while the white spot and the peripheral vocal cord tissue GPRS 30 are mainly located in the nucleus .

3 . gprs 30 may be involved in the vocal cord inflammatory response of women in menstrual cycles due to e2 fluctuations .
【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R767

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