牙髓炎癥誘導(dǎo)P2X受體家族在外周及中樞神經(jīng)系統(tǒng)中的表達(dá)
發(fā)布時(shí)間：2019-02-13 05:42
【摘要】：第一部分LPS誘導(dǎo)牙髓炎癥過程中P2X受體在三叉神經(jīng)節(jié)中的表達(dá)和作用 研究目的：離子通道型嘌呤受體家族(P2X)可以參與調(diào)節(jié)初級感覺神經(jīng)元中的疼痛感覺。本實(shí)驗(yàn)?zāi)康脑谟谘芯吭谘浪柩装Y引起三叉神經(jīng)節(jié)(trigeminal ganglion,TG)中神經(jīng)元活化以及P2X受體的表達(dá)模式,并采用P2X受體抑制劑觀察抑制P2X受體后對痛覺傳導(dǎo)相關(guān)因子表達(dá)的改變的影響。 研究方法：本實(shí)驗(yàn)將脂多糖(Lipopolysaccharides,LPS)置于大鼠左側(cè)的上頜第一磨牙髓腔,以誘導(dǎo)牙髓炎癥。觀察了急性牙髓炎癥期TG中活化神經(jīng)元標(biāo)志c-fos的分布,P2X受體與谷氨酸能神經(jīng)元和GABA能神經(jīng)元的關(guān)系,并采用免疫組化和蛋白印跡技術(shù)觀察了TG的V1-V2支中的P2X受體的表達(dá)模式。進(jìn)一步在髓腔中置入P2X受體抑制劑TNP-ATP并同時(shí)于大鼠左側(cè)須墊注射TNP-ATP,觀察了抑制P2X受體對急性牙髓炎癥期TG中c-fos,谷氨酸能神經(jīng)元標(biāo)志VGluT1和GABA能神經(jīng)元標(biāo)志GAD67表達(dá),以及p38和ERK1/2磷酸化的影響。 研究結(jié)果：FG逆行性標(biāo)記顯示同側(cè)TG的V1-V2支支配左上頜第一磨牙,牙髓炎癥可導(dǎo)致V1-V2支中神經(jīng)元活化的標(biāo)志c-fos表達(dá)增加；P2X2, P2X3和P2X5受體在TG的神經(jīng)元胞漿中表達(dá),在牙髓炎癥進(jìn)展的第1天表達(dá)增加,至第三天持續(xù)增加；P2X2, P2X3和P2X5受體在V1-V2支中的表達(dá)增高主要發(fā)生在與痛覺傳導(dǎo)相關(guān)的小直徑神經(jīng)元中。熒光雙標(biāo)記法顯示P2X2,P2X3和P2X5受體在V1-V2支中分別與VGluT1和GAD67共表達(dá)在小直徑的TG神經(jīng)元的胞漿中；LPS刺激牙髓誘導(dǎo)炎癥能導(dǎo)致c-fos, VGluT1和GAD67表達(dá)增高,并導(dǎo)致p-p38和p-ERK1/2表達(dá)增加；抑制P2X受體會(huì)導(dǎo)致c-fos, VGluT1和GAD67的表達(dá)增高受到抑制,并降低p-p38和p-ERK1/2表達(dá)。 研究結(jié)論：LPS誘導(dǎo)的急性牙髓炎癥可誘導(dǎo)P2X2, P2X3和P2X5受體在活化的TGV1-V2支中的表達(dá)增高,提示P2X受體可能參與了牙髓炎癥誘導(dǎo)的神經(jīng)敏感性增高。抑制P2X受體會(huì)使牙髓炎癥引起的TG中c-fos,VGluT1和GAD67的表達(dá)增高受到抑制,并降低p38和ERK1/2的磷酸化。 第二部分牙髓炎癥對大鼠中樞神經(jīng)系統(tǒng)中P2X表達(dá)的影響 研究目的：本實(shí)驗(yàn)擬采用大鼠開髓模型誘導(dǎo)大鼠牙髓炎癥,目的在于研究不同牙髓炎癥進(jìn)展階段對腦干的三叉神經(jīng)脊束核尾端亞核(Vc)和丘腦的腹后中核(VPM)中的影響。 研究方法：本實(shí)驗(yàn)在大鼠左上頜第一磨牙頜面開髓,并暴露與口腔菌群環(huán)境,誘導(dǎo)牙髓炎癥。取牙髓炎癥第1天,第3天,第7天和第28天時(shí),大鼠雙側(cè)腦干的三叉神經(jīng)脊束核尾端亞核(Vc)和丘腦的腹后中核(VPM),采用免疫組化和免疫熒光方法觀察P2X受體家族(P2X1-7受體)的表達(dá)模式。 研究結(jié)果：實(shí)驗(yàn)觀察到牙髓炎癥第1天,第3天,第7天顯示急性牙髓炎癥的組織學(xué)表現(xiàn),其中第3天最為典型,而第28天顯示牙髓壞死和慢性根尖周炎的組織學(xué)表現(xiàn)；P2X1-6受體在Vc和VPM中均表達(dá)在神經(jīng)元的胞漿中,而P2X7表達(dá)在Vc和VPM中在小膠質(zhì)細(xì)胞中；P2X2,P2X4和P2X7受體在急性牙髓炎癥期在同側(cè)Vc中的表達(dá)增加；P2X1-5和P2X7的表達(dá)都在雙側(cè)VPM中均上升,開髓3d時(shí),對側(cè)VPM中P2X1-5和P2X7的平均表達(dá)面積顯著高于同側(cè)。 研究結(jié)論：除P2X6受體持續(xù)高表達(dá)外,P2X1-5和P2X7受體均可受到牙髓炎癥的誘導(dǎo),在Vc和VPM中表達(dá)增高。
[Abstract]:The expression and function of P2X receptor in the trigeminal ganglion during the first part of LPS-induced dental pulp inflammation The purpose of this study is that the ion channel type methotrexate receptor family (P2X) can be involved in the regulation of the pain in primary sensory neurons. The purpose of this study was to study the activation of the neurons in the trigeminal ganglion (TG) and the expression pattern of the P2X receptor in the pulpal inflammation, and to use the P2X receptor inhibitor to observe the effect of the P2X receptor on the expression of the hyperalgesia-related factors. In this experiment, Lipopolysaccharide (LPS) was placed on the left side of the rat's maxillary first pulp cavity to induce the teeth. The distribution of c-fos, the distribution of the c-fos, the relationship between the P2X receptor and the neuronal and GABA-ergic neurons in the TG of acute endodontic inflammation were observed, and the expression of the P2X receptor in the V1-V2 branch of TG was observed by immunohistochemistry and Western blotting. In the model, the P2X receptor inhibitor TNP-ATP was further placed in the medullary cavity and the TNP-ATP was injected at the left side of the rat, and the expression of the c-fos, the glutamate-capable neuronal marker VGluT1 and the GABA-enabled neuronal marker GAD67 in the acute endodontic inflammation phase TG, and the phosphorylation of p38 and ERK1/ 2 were observed. The results of the study: The reverse marker of FG showed that the V1-V2 branch of the same side was dominant to the left maxillary first molar, which could lead to the increase of the expression of c-fos in the neurons in the V1-V2 branch, and the expression of the P2X2, P2X3 and P2X5 receptors in the cytoplasm of the neurons in the TG, and the first in the progress of the dental pulp inflammation. The increase of the expression of P2X2, P2X3 and P2X5 in the V1-V2 branch mainly occurred in the small straight line related to the conduction of the hyperalgesia. The expression of P2X2, P2X3 and P2X5 was co-expressed with VGluT1 and GAD67 in the cytoplasm of the small-diameter TG neurons, and the expression of p-p38 and p-ERK1/ 2 increased and the expression of p-p38 and p-ERK1/ 2 increased, and the inhibition of P2X receptors could lead to increased expression of p-p38 and p-ERK1/ 2. The expression of c-fos, VGluT1 and GAD67 was inhibited and the p-p38 and p-ERK were reduced. Conclusion: The expression of P2X2, P2X3 and P2X5 receptor in the activated TGV1-V2 branch can be induced by LPS-induced acute dental pulp inflammation, suggesting that the P2X receptor may be involved in the induction of dental pulp inflammation. The inhibition of the expression of c-fos, VGluT1 and GAD67 in the TG induced by the pulpal inflammation by the P2X receptor is inhibited and p38 and ERK are reduced. 1/ 2 phosphorylation. The second part of dental pulp inflammation is the central nervous system of the rat The effect of P2X expression in the system is the purpose of the study: this experiment is to be used in rats The purpose of this study was to study the subnucleus (Vc) of the nucleus of the trigeminal nerve of the brain stem and the ventral part of the thalamus in the stage of different pulp inflammation. The effect of this experiment on the maxillary first molar and maxillofacial opening of the rat's left maxillary first molar and the exposure to The first day, third day, day 7 and day 28 of the dental pulp inflammation, the subnucleus (Vc) and the nucleus (VPM) of the posterior subnucleus (VPM) of the nucleus of the trigeminal nerve of the rat's dual-side brain stem were observed, and the P2X receptor family (P) was observed by the method of immunohistochemistry and immunofluorescence. The expression pattern of the 2X1-7 receptor was observed. The results of the study showed that the first day, the third day and the seventh day of the dental pulp inflammation showed the histological appearance of the acute dental pulp inflammation, in which the third day was the most typical, and the dental pulp was shown on the 28th day. The expression of P2X2, P2X4, and P2X7 in the same side of Vc and VPM increased with the expression of P2X2, P2X4, and P2X7 in the same side, and the expression of P2X1-5 and P2X7 was in the same side. Both sides VPM rise and open the nucleus for 3d, P2X1-5 and P2 in the side VPM The average expression area of X7 was significantly higher than that of the same side. Conclusion: In addition to the high expression of P2X6 receptor, P2X1-5 and P2X7 receptors can be affected by pulpitis.
【學(xué)位授予單位】：武漢大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2014
【分類號】：R781.31
本文編號：2421235
[Abstract]:The expression and function of P2X receptor in the trigeminal ganglion during the first part of LPS-induced dental pulp inflammation The purpose of this study is that the ion channel type methotrexate receptor family (P2X) can be involved in the regulation of the pain in primary sensory neurons. The purpose of this study was to study the activation of the neurons in the trigeminal ganglion (TG) and the expression pattern of the P2X receptor in the pulpal inflammation, and to use the P2X receptor inhibitor to observe the effect of the P2X receptor on the expression of the hyperalgesia-related factors. In this experiment, Lipopolysaccharide (LPS) was placed on the left side of the rat's maxillary first pulp cavity to induce the teeth. The distribution of c-fos, the distribution of the c-fos, the relationship between the P2X receptor and the neuronal and GABA-ergic neurons in the TG of acute endodontic inflammation were observed, and the expression of the P2X receptor in the V1-V2 branch of TG was observed by immunohistochemistry and Western blotting. In the model, the P2X receptor inhibitor TNP-ATP was further placed in the medullary cavity and the TNP-ATP was injected at the left side of the rat, and the expression of the c-fos, the glutamate-capable neuronal marker VGluT1 and the GABA-enabled neuronal marker GAD67 in the acute endodontic inflammation phase TG, and the phosphorylation of p38 and ERK1/ 2 were observed. The results of the study: The reverse marker of FG showed that the V1-V2 branch of the same side was dominant to the left maxillary first molar, which could lead to the increase of the expression of c-fos in the neurons in the V1-V2 branch, and the expression of the P2X2, P2X3 and P2X5 receptors in the cytoplasm of the neurons in the TG, and the first in the progress of the dental pulp inflammation. The increase of the expression of P2X2, P2X3 and P2X5 in the V1-V2 branch mainly occurred in the small straight line related to the conduction of the hyperalgesia. The expression of P2X2, P2X3 and P2X5 was co-expressed with VGluT1 and GAD67 in the cytoplasm of the small-diameter TG neurons, and the expression of p-p38 and p-ERK1/ 2 increased and the expression of p-p38 and p-ERK1/ 2 increased, and the inhibition of P2X receptors could lead to increased expression of p-p38 and p-ERK1/ 2. The expression of c-fos, VGluT1 and GAD67 was inhibited and the p-p38 and p-ERK were reduced. Conclusion: The expression of P2X2, P2X3 and P2X5 receptor in the activated TGV1-V2 branch can be induced by LPS-induced acute dental pulp inflammation, suggesting that the P2X receptor may be involved in the induction of dental pulp inflammation. The inhibition of the expression of c-fos, VGluT1 and GAD67 in the TG induced by the pulpal inflammation by the P2X receptor is inhibited and p38 and ERK are reduced. 1/ 2 phosphorylation. The second part of dental pulp inflammation is the central nervous system of the rat The effect of P2X expression in the system is the purpose of the study: this experiment is to be used in rats The purpose of this study was to study the subnucleus (Vc) of the nucleus of the trigeminal nerve of the brain stem and the ventral part of the thalamus in the stage of different pulp inflammation. The effect of this experiment on the maxillary first molar and maxillofacial opening of the rat's left maxillary first molar and the exposure to The first day, third day, day 7 and day 28 of the dental pulp inflammation, the subnucleus (Vc) and the nucleus (VPM) of the posterior subnucleus (VPM) of the nucleus of the trigeminal nerve of the rat's dual-side brain stem were observed, and the P2X receptor family (P) was observed by the method of immunohistochemistry and immunofluorescence. The expression pattern of the 2X1-7 receptor was observed. The results of the study showed that the first day, the third day and the seventh day of the dental pulp inflammation showed the histological appearance of the acute dental pulp inflammation, in which the third day was the most typical, and the dental pulp was shown on the 28th day. The expression of P2X2, P2X4, and P2X7 in the same side of Vc and VPM increased with the expression of P2X2, P2X4, and P2X7 in the same side, and the expression of P2X1-5 and P2X7 was in the same side. Both sides VPM rise and open the nucleus for 3d, P2X1-5 and P2 in the side VPM The average expression area of X7 was significantly higher than that of the same side. Conclusion: In addition to the high expression of P2X6 receptor, P2X1-5 and P2X7 receptors can be affected by pulpitis.
【學(xué)位授予單位】：武漢大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2014
【分類號】：R781.31
【參考文獻(xiàn)】
相關(guān)期刊論文 前1條
1 劉宇煒;陳曉青;田香;陳琳;吳鈺祥;黃丹;易卉玲;易初麗;李超英;;P2X_3 , but not P2X_1 , Receptors Mediate ATP-activated Current in Neurons Innervating Tooth-pulp[J];Journal of Huazhong University of Science and Technology(Medical Sciences);2013年03期
，本文編號：2421235
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