本文選題：妊娠期煙熏　切入點(diǎn)：新生大鼠　出處：《新鄉(xiāng)醫(yī)學(xué)院》2014年碩士論文

【摘要】：背景眾所周知吸煙成癮已成為這個(gè)社會不可忽視的問題,抽煙不僅對吸煙者本人乃至周圍的人甚至子代都產(chǎn)生不良影響。孕期尼古丁暴露會導(dǎo)致胎兒造成極大的危害,如：致畸、宮內(nèi)缺氧、低體重兒、先天性疾病、嬰兒猝死癥候群等。煙堿是香煙中的主要生物堿,在妊娠期煙堿暴露所致胎兒中樞性呼吸異常中起關(guān)鍵作用,而穩(wěn)定的節(jié)律性呼吸對于維持機(jī)體正常的生命活動(dòng)至關(guān)重要。因此,研究孕期尼古丁暴露對新生大鼠延髓腦片基本節(jié)律性放電和在體耗氧量的作用至關(guān)重要,為進(jìn)一步明確孕期尼古丁暴露對子代延髓呼吸中樞的抑制作用,并為相關(guān)疾病的預(yù)防和治療提供新思路和實(shí)驗(yàn)基礎(chǔ)。 目的研究妊娠期煙熏對子代延髓呼吸中樞基本節(jié)律性呼吸放電及延髓呼吸中樞a7nACh受體(a7nicotinic acetylcholinergic receptor, a7nAChR)蛋白和基因表達(dá)的變化,探討α7nAChR在孕期煙熏中所致子代呼吸抑制中的作用和機(jī)制。 方法0-3天Sprague-Dawley大鼠,雌雄不拘,隨機(jī)分為對照組和實(shí)驗(yàn)組。對照組新生大鼠產(chǎn)自自然日夜規(guī)律、常規(guī)飼養(yǎng)的成年大鼠；實(shí)驗(yàn)組新生大鼠產(chǎn)自除常規(guī)飼養(yǎng)外,自合籠至分娩每日2次(8Am和8Pm)、置于80cm*60cm*100cm*煙熏箱中8只香煙煙熏30min的大鼠。 1.用新生大鼠離體延髓腦片標(biāo)本記錄反映延髓呼吸中樞功能的腦片基本節(jié)律性呼吸放電(rhythmic respiratory discharge activity, RRDA)。觀察α7nAChR激動(dòng)劑乙酰膽堿(acetylcholine,ACh)和拮抗劑銀環(huán)蛇毒素(Alpha-bungarotoxin,α-BGT)對對照組和實(shí)驗(yàn)組RRDA的作用,分析α7nAChR是否對RRDA有抑制作用,以明確α7nAChR是否參與孕期煙熏對子代的呼吸抑制作用。 2.采用Western Blot技術(shù)檢測α7nAChR在延髓呼吸中樞面神經(jīng)后核內(nèi)側(cè)區(qū)神經(jīng)元上的表達(dá),明確孕期煙熏是否減少α7nAChR在延髓呼吸中樞面神經(jīng)后核內(nèi)側(cè)區(qū)神經(jīng)元上的表達(dá)。 3.采用qRT-PCR技術(shù)檢測α7nAChR mRNA在延髓呼吸中樞面神經(jīng)后核內(nèi)側(cè)區(qū)神經(jīng)元上的表達(dá),明確孕期煙熏是否減少α7nAChR mRNA在延髓呼吸中樞面神經(jīng)后核內(nèi)側(cè)區(qū)神經(jīng)元上的表達(dá)。分析α7nAChR功能改變與受體和αRNA表達(dá)是否一致,是否存在相關(guān)性。 結(jié)果 1.對照組和實(shí)驗(yàn)組腦片放電60min內(nèi)穩(wěn)定無衰減,實(shí)驗(yàn)?zāi)Ｐ头�(wěn)定可靠。實(shí)驗(yàn)組腦片放電弱于對照組(以對照組為100%,實(shí)驗(yàn)組20min時(shí)TI88.23±3.17%、IA89.52±4.74%、RC90.06±4.57%)；α7nAChR激動(dòng)劑ACh對實(shí)驗(yàn)組和對照組腦片放電都有興奮作用,對照組興奮效果強(qiáng)于實(shí)驗(yàn)組(實(shí)驗(yàn)組變化率分別13.05±0.46%、10.37±0.49%、7.65±±±0.40%；對照組為：19.67±0.54%、17.55±0.49%18.04±0.58%)；α7nAChR拮抗劑a-BGT對實(shí)驗(yàn)組和對照組腦片放電都有抑制作用,對照組抑制效果強(qiáng)于實(shí)驗(yàn)組(實(shí)驗(yàn)組變化率分別為9.67±0.50%、8.43±±±0.49%、6.67±0.53%;對照組為：16.65±0.45%、14.92±0.49%、14.33±0.43%)。 2. Western Blot實(shí)驗(yàn)結(jié)果顯示,與對照組相比,實(shí)驗(yàn)組α7nAChR在延髓呼吸中樞面神經(jīng)后核內(nèi)側(cè)區(qū)神經(jīng)元表達(dá)減少(對照組1.012±0.090、實(shí)驗(yàn)組0.604±0.034)。 3qRT-PCR實(shí)驗(yàn)結(jié)果顯示,與對照組相比,實(shí)驗(yàn)組α7nAChRmRNA在延髓呼吸中樞面神經(jīng)后核內(nèi)側(cè)區(qū)神經(jīng)元表達(dá)水平下調(diào)(對照組1.004±0.101、實(shí)驗(yàn)組0.809±0.069)。 結(jié)論 1.孕期煙熏抑制新生鼠延髓腦片基本節(jié)律性呼吸放電、抑制α7nAChR對RRDA的調(diào)節(jié)作用,下調(diào)α7nACh受體蛋白和mRNA表達(dá)水平。 2.抑制α7nACh受體活性及功能,通過下調(diào)α7nACh受體蛋白水平和受體mRNA表達(dá)水平使延髓呼吸中樞面神經(jīng)后核內(nèi)側(cè)區(qū)神經(jīng)元數(shù)量減少、功能下降可能是孕期煙熏抑制子代延髓中樞呼吸功能的機(jī)制之一。
[Abstract]:Background : It is well known that smoking addiction has become a non - negligible problem in society . Smoking is not only harmful to smokers themselves or even those around them . The exposure to nicotine in pregnancy plays a key role in fetal central respiratory abnormalities due to exposure to nicotine .

Objective To study the effect and mechanism of 偽 - 7nCR on the respiratory depression in children during pregnancy .

Methods Sprague - Dawley rats were randomly divided into two groups : control group and experimental group .
In the experimental group , the newborn rats were fed twice daily ( 8Am and 8Pm ) except for routine feeding , and were placed in 80 cm * 60 cm * 100 cm * smoking oven for 30 min .

1 . The basic rhythmic respiratory discharge activity ( RRDA ) of the brain slices reflecting the function of the medullary respiratory center was recorded from the specimen of the isolated medulla of the neonatal rat . The effect of 偽 - bungarotoxin ( 偽 - BGT ) on RRDA in the control group and experimental group was investigated .

2 . Western Blot was used to detect the expression of 偽7nCR on the neurons in the medial region of the nucleus of the central facial nerve of the medullary respiratory center .

3 . Using qRT - PCR technique to detect the expression of 偽7nCR mRNA in the medial region of the nucleus of the central facial nerve of the medullary respiratory center .

Results

1 . The experimental model was stable and reliable in the control group and the experimental group , and the experimental model was stable and reliable . The experimental group was weaker than the control group ( 100 % in the control group , TI88.23 鹵 3.17 % , IA89.52 鹵 4.74 % , RC90.06 鹵 4.57 % ) .
In the experimental group and the control group , there was an exciting effect on both the experimental group and the control group . The control group was more excited than the experimental group ( 13.05 鹵 0.46 % , 10.37 鹵 0.49 % , 7.65 鹵 鹵 0.40 % , respectively ) .
The control group was 19.67 鹵 0.54 % , 17.55 鹵 0.49 % 18.04 鹵 0.58 % ;
The results showed that a - BGT inhibited the discharges of brain slices in the experimental group and the control group , and the control group was stronger than the experimental group ( 9.67 鹵 0.50 % , 8.43 鹵 鹵 0.49 % , 6.67 鹵 0.53 % , 14.92 鹵 0.49 % , 14.33 鹵 0.43 % ) .

2 . Compared with the control group , the expression of 偽 - 7nR _ ( R ) R _ ( R ) R _ ( R _ ( R ) R _ ( R ) _ ( R _ ( R ) R _ ( R ) _ ( R _ ( R ) R _ ( R ) _ ( R _ ( R ) _ ( R ) _ ( R _ ( R ) _ ( R ) _ ( R _ ( R ) _ ( R ) _ ( R _ ( R ) _ ( R ) _ ( R _ ( R ) _ ( R ) _ ( R _ ( R ) _ ( R ) _ ( R _ ( R ) _ ( R ) _ ( R _ ( R ) _ ( R ) _ ( R ) in the

The results of 3qRT - PCR showed that the expression of 偽 - 7nCDRmRNA in experimental group was lower than that of control group ( 1.004 鹵 0.101 in the control group and 0.809 鹵 0.069 in the experimental group ) .

Conclusion

1 . During the pregnancy period , the basic rhythmic respiratory discharge was inhibited by smoke suppression in neonatal rats , and the regulatory action of 偽7nCR on RRDA was inhibited , and the expression level of 偽7nAl _ 1 receptor protein and mRNA was downregulated .

2 . The inhibition of the activity and function of 偽 - 7nach receptor , the decrease of the number of neurons in the medial region of the nucleus of the nucleus of the medullary respiratory center after downregulation of the level of the 偽 - 7nabolol receptor protein and the level of receptor mRNA , the decrease in function may be one of the mechanisms for the inhibition of the respiratory function of the nucleus of the medulla of the subnucleus during pregnancy .

【學(xué)位授予單位】：新鄉(xiāng)醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R714.2

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