中华口腔医学杂志
中華口腔醫學雜誌
중화구강의학잡지
Chinese Journal of Stomatology
2013年
6期
330-333
,共4页
陈旭%张珍丽%孙鲁雁%郭艳
陳旭%張珍麗%孫魯雁%郭豔
진욱%장진려%손로안%곽염
牙髓%牙乳头%LIM矿化蛋白1%年轻恒牙
牙髓%牙乳頭%LIM礦化蛋白1%年輕恆牙
아수%아유두%LIM광화단백1%년경항아
Dental pulp%Dental papilla%LIM mineralization protein-1%Immature permanent teeth
目的 比较人年轻恒牙牙髓、根尖牙乳头及成熟恒牙牙髓组织中LIM矿化蛋白1(LIMmineralization protein-1,LMP-1)的表达,探寻LMP-1在牙髓牙本质复合体发育和成熟中的作用.方法 收集48颗因正畸拔除的健康前磨牙,其中年轻和成熟恒牙各24颗,样本来源对象的年龄分别为11 ～14岁和18 ～30岁.拔除后即刻分离获得年轻恒牙的牙髓和根尖牙乳头组织(各24个样本)以及成熟恒牙的牙髓组织(24个样本).实验分组如下:第1组为牙根形成2/3的年轻恒牙的根尖牙乳头组织；第2组为牙根形成2/3的年轻恒牙的牙髓组织；第3组为成熟恒牙的牙髓组织.每组12个样本用于反转录PCR(reverse transcription-PCR,RT-PCR)检测,另12个样本用于蛋白质印迹法检测,以β-肌动蛋白做内参.图像分析软件(Meta Morph 6.2.6)对阳性条带进行灰度值测定,目的条带与内参的灰度值比值为LMP-1的表达强度.应用SPSS13.0对数据进行统计学分析,LMP-1在年轻恒牙根尖牙乳头与牙髓组织之间的表达强度的两两比较采用两配对样本的t检验,年轻恒牙牙髓与成熟恒牙牙髓组织之间的表达强度的两两比较采用两独立样本的t检验,以双侧P＜0.05为差异有统计学意义.结果 LMP-1 mRNA在年轻恒牙的根尖牙乳头、牙髓和成熟恒牙的牙髓组织中的表达强度分别为0.25±0.09、0.46±0.24和0.31 ±0.10,经两样本t检验分析,LMP-1的基因表达在第2组年轻恒牙牙髓组织明显高于其在第1组年轻恒牙根尖牙乳头组织(t=2.92)以及第3组成熟恒牙牙髓组织的表达强度(t=2.31,P＜0.05).LMP-1蛋白在3组中的表达强度分别为0.33 ±0.08、0.82±0.10和0.52 ±0.19,LMP-1的蛋白表达在第2组显著高于第1组(t=3.33)及第3组中的表达强度(t =3.11,P＜0.05).结论 无论是在mRNA水平还是在蛋白水平,LMP-1在人年轻恒牙牙髓、根尖牙乳头以及成熟恒牙牙髓组织中均有不同程度的表达,提示LMP-1在牙髓牙本质复合体发育和成熟过程中可能发挥重要作用.
目的 比較人年輕恆牙牙髓、根尖牙乳頭及成熟恆牙牙髓組織中LIM礦化蛋白1(LIMmineralization protein-1,LMP-1)的錶達,探尋LMP-1在牙髓牙本質複閤體髮育和成熟中的作用.方法 收集48顆因正畸拔除的健康前磨牙,其中年輕和成熟恆牙各24顆,樣本來源對象的年齡分彆為11 ～14歲和18 ～30歲.拔除後即刻分離穫得年輕恆牙的牙髓和根尖牙乳頭組織(各24箇樣本)以及成熟恆牙的牙髓組織(24箇樣本).實驗分組如下:第1組為牙根形成2/3的年輕恆牙的根尖牙乳頭組織；第2組為牙根形成2/3的年輕恆牙的牙髓組織；第3組為成熟恆牙的牙髓組織.每組12箇樣本用于反轉錄PCR(reverse transcription-PCR,RT-PCR)檢測,另12箇樣本用于蛋白質印跡法檢測,以β-肌動蛋白做內參.圖像分析軟件(Meta Morph 6.2.6)對暘性條帶進行灰度值測定,目的條帶與內參的灰度值比值為LMP-1的錶達彊度.應用SPSS13.0對數據進行統計學分析,LMP-1在年輕恆牙根尖牙乳頭與牙髓組織之間的錶達彊度的兩兩比較採用兩配對樣本的t檢驗,年輕恆牙牙髓與成熟恆牙牙髓組織之間的錶達彊度的兩兩比較採用兩獨立樣本的t檢驗,以雙側P＜0.05為差異有統計學意義.結果 LMP-1 mRNA在年輕恆牙的根尖牙乳頭、牙髓和成熟恆牙的牙髓組織中的錶達彊度分彆為0.25±0.09、0.46±0.24和0.31 ±0.10,經兩樣本t檢驗分析,LMP-1的基因錶達在第2組年輕恆牙牙髓組織明顯高于其在第1組年輕恆牙根尖牙乳頭組織(t=2.92)以及第3組成熟恆牙牙髓組織的錶達彊度(t=2.31,P＜0.05).LMP-1蛋白在3組中的錶達彊度分彆為0.33 ±0.08、0.82±0.10和0.52 ±0.19,LMP-1的蛋白錶達在第2組顯著高于第1組(t=3.33)及第3組中的錶達彊度(t =3.11,P＜0.05).結論 無論是在mRNA水平還是在蛋白水平,LMP-1在人年輕恆牙牙髓、根尖牙乳頭以及成熟恆牙牙髓組織中均有不同程度的錶達,提示LMP-1在牙髓牙本質複閤體髮育和成熟過程中可能髮揮重要作用.
목적 비교인년경항아아수、근첨아유두급성숙항아아수조직중LIM광화단백1(LIMmineralization protein-1,LMP-1)적표체,탐심LMP-1재아수아본질복합체발육화성숙중적작용.방법 수집48과인정기발제적건강전마아,기중년경화성숙항아각24과,양본래원대상적년령분별위11 ～14세화18 ～30세.발제후즉각분리획득년경항아적아수화근첨아유두조직(각24개양본)이급성숙항아적아수조직(24개양본).실험분조여하:제1조위아근형성2/3적년경항아적근첨아유두조직；제2조위아근형성2/3적년경항아적아수조직；제3조위성숙항아적아수조직.매조12개양본용우반전록PCR(reverse transcription-PCR,RT-PCR)검측,령12개양본용우단백질인적법검측,이β-기동단백주내삼.도상분석연건(Meta Morph 6.2.6)대양성조대진행회도치측정,목적조대여내삼적회도치비치위LMP-1적표체강도.응용SPSS13.0대수거진행통계학분석,LMP-1재년경항아근첨아유두여아수조직지간적표체강도적량량비교채용량배대양본적t검험,년경항아아수여성숙항아아수조직지간적표체강도적량량비교채용량독립양본적t검험,이쌍측P＜0.05위차이유통계학의의.결과 LMP-1 mRNA재년경항아적근첨아유두、아수화성숙항아적아수조직중적표체강도분별위0.25±0.09、0.46±0.24화0.31 ±0.10,경량양본t검험분석,LMP-1적기인표체재제2조년경항아아수조직명현고우기재제1조년경항아근첨아유두조직(t=2.92)이급제3조성숙항아아수조직적표체강도(t=2.31,P＜0.05).LMP-1단백재3조중적표체강도분별위0.33 ±0.08、0.82±0.10화0.52 ±0.19,LMP-1적단백표체재제2조현저고우제1조(t=3.33)급제3조중적표체강도(t =3.11,P＜0.05).결론 무론시재mRNA수평환시재단백수평,LMP-1재인년경항아아수、근첨아유두이급성숙항아아수조직중균유불동정도적표체,제시LMP-1재아수아본질복합체발육화성숙과정중가능발휘중요작용.
Objective To examine the expression of LIM mineralization protein-1 (LMP-1) in the apical papilla and dental pulp tissues of human immature permanent teeth and to investigate the role of LMP-1 in the development and maturation of pulp-dentin complex.Methods Forty-eight healthy premolars in need of extraction for orthodontic treatment were obtained with 24 immature permanent teeth and 24 mature permanent teeth.After extraction,the apical papilla was detached from the dental pulp in the immature permanent tooth and the dental pulp of mature permanent tooth was rapidly removed.The samples were divided into 3 groups:group 1,apical papilla of immature permanent teeth (root formed 2/3 of its full length) ; group 2,dental pulp tissues of immature permanent teeth; group 3,dental pulp tissues of mature permanent teeth.There were 24 samples for each group.Half of them were used for reverse transcriptien-PCR (RT-PCR) detection,and the other half were used for Western blotting detection.Band intensities were quantified using Meta Morph software 6.2.6 and subsequently normalized by dividing the band gray value ofthe target gene by the intensity of its corresponding β-actin.Two-sample t test was used to analyze the difference of expression intensity between group 1 and group 2 as well as group 2 and group 3 with SPSS 13.0 software package.Statistical significance was established as P ＜ 0.05.Results As indicated by RT-PCR,LMP-1 expressed in the apical papilla,dental pulp of immature permanent teeth and the dental pulp of mature permanent teeth were 0.25 ± 0.09,0.46 ± 0.24 and 0.31 ± 0.10 respectively.The expression intensity of LMP-1 mRNA in the dental pulp tissues of human immature permanent teeth were significantly higher than that in the apical papilla tissues(t =2.92) and that in the dental pulp tissues of human mature permanent teeth(t =2.31) (P ＜ 0.05).As indicated by Western blotting,LMP-1 expressed in the three groups were 0.33 ± 0.08,0.82 ± 0.10 and 0.52 ± 0.19 respectively.The expression intensity of LMP-1 protein in the dental pulp tissues of human immature permanent teeth were significantly higher than that in the apical papilla tissues(t =3.33) and that in the dental pulp tissues of human mature permanent teeth(t =3.11)(P ＜ 0.05).Conclusions LMP-1 were positively expressed in all the samples including apical papilla of immature permanent teeth,dental pulp of immature and mature permanent teeth at the level of both mRNA and protein,but with different intensity.LMP-1 could play an important role in the development and maturation of pulp-dentin complex.
