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目的探讨大鼠牙胚发育过程中Biglycan的表达情况。方法健康雌性和雄性SD大鼠各15只,按雌∶雄=1∶1同笼饲养,精子涂片检查阳性时为受孕,按取材时间将15只孕鼠分为妊娠17.5d组、妊娠19.5d组、出生后1.5d组、出生后5.5d组和出生后9.5d组,每组3只。分别在妊娠第17.5天、第19.5天时取胎鼠及在出生后第1.5、5.5、9.5天时取新生鼠两侧下颌第一磨牙牙胚部位后的下颌骨组织,采用组织病理检查确定各组大鼠牙胚发育时期,采用免疫组织化学法检测大鼠牙胚Biglycan蛋白多糖的表达和定位情况,采用图像分析软件分析各组Biglycan蛋白多糖的累计光密度、有效目标分布区域面积和平均光密度。结果妊娠17.5d组胎鼠第一磨牙牙胚处于帽状期,Biglycan表达情况为牙板细胞(+)、牙囊细胞(■)、内釉上皮(■)及牙乳头细胞(■);妊娠19.5d组处于钟状早期,Biglycan表达情况同妊娠17.5d组;出生后1.5d组处于钟状期,Biglycan表达情况为压板细胞(■)、牙囊细胞(+)、内釉上皮(+)及牙乳头细胞(+);出生后5.5d组处于钟状末期,Biglycan表达情况为牙囊细胞(+)、内釉上皮(+)及牙乳头细胞(±);出生后9.5d组第一磨牙牙胚釉基质蛋白分泌基本结束,Biglycan表达情况为压板细胞(±)、牙囊细胞(+)及牙乳头细胞(±);随着发育进行,成釉细胞和成牙本质细胞中Biglycan表达逐渐减弱;妊娠17.5d组Biglycan蛋白多糖累计光密度值为1.470 0±0.001 1,有效目标分布区域面积为4.323 5±0.001 6,平均光密度值为0.340 0±0.002 3,妊娠19.5d组依次为1.460 0±0.001 7,4.424 2±0.001 5,0.034 0±0.004 5,出生后1.5d组依次为1.440 0±0.001 4,4.363 6±0.001 3,0.310 0±0.002 2,出生后5.5d组依次为1.430 0±0.001 3,4.612 9±0.001 1,0.300 0±0.000 1,出生后9.5d组依次为1.410 0±0.001 5,7.421 1±0.001 4,0.190 0±0.002 8,各组间两两比较差异均有统计学意义(P<0.05)。结论正常大鼠牙胚发育过程中Biglycan表达呈一定的时间和空间分布,随着发育的不同阶段而有所改变,Biglycan可能参与了牙胚的正常发育。
Objective To investigate the expression of Biglycan in rat tooth germ development. Methods Fifteen healthy female and male Sprague-Dawley rats were housed in the same cage according to the ratio of female to male with 1: 1. When pregnant with sperm smear positive test, 15 pregnant rats were divided into 17.5 d of pregnancy and 19.5 d of pregnancy d group, 1.5d after birth, 5.5d after birth and 9.5d after birth, three in each group. The fetuses were taken from 17.5 days and 19.5 days of gestation and the mandibular tissues were taken from the first molar on both sides of the newborn rat on the first, fifth, fifth and fifth days after birth respectively. At the period of mouse tooth germ development, the expression and localization of Biglycan proteoglycan in rat tooth germ were detected by immunohistochemical method. The total optical density, effective target distribution area and average optical density of Biglycan proteoglycan in each group were analyzed by image analysis software. Results The first molar tooth germ in 17.5 d gestation group was in cap-like shape, and the expression of Biglycan was (+), dental follicle cells (■), endometrial epithelium (■) and papillary cells (■) The expression of Biglycan in the bell-shaped early stage was similar to that of the 17.5-day gestation group in the group of 19.5d. The expression of Biglycan in the bell-shaped stage was 1.5 days after birth. The expression of Biglycan was (+), (+), Dental papilla cells (+) and dental papilla cells (+). The postnatal 5.5d group was in the bell-shaped end stage. The expression of Biglycan was (+ The expression of Biglycan in placental cells (±), dental follicle cells (+) and papillary cells (±) were significantly reduced after tooth germ formation. The expression of Biglycan in ameloblasts and odontoblasts The cumulative optical density of Biglycan proteoglycan in pregnancy 17.5d group was 1.470 0 ± 0.001 1, the effective target distribution area was 4.323 5 ± 0.001 6, the average optical density value was 0.340 0 ± 0.002 3, and the group of 19.5d gestation was 1.460 0 ± 0.001 7,4.424 2 ± 0.001 5,0.034 0 ± 0.004 5 1.5d after birth were 1.440 0 ± 0.001 4,4.363 6 ± 0.001 3, 0.310 0 ± 0.002 2 after birth 5.5d group followed by 1.430 0 ± 0.001 3,4.612 9 ± 0.001 1, 0.300 0 ± 0.000 1 9.5d after birth were 1.410 0 ± 0.001 5,7.421 1 ± 0.001 4, 0.190 0 ± 0.002 8, the difference between every two groups was statistically significant (P <0.05). Conclusion Biglycan expression in normal rat tooth germ has some time and space distribution, which changes with different stages of development. Biglycan may be involved in the normal development of tooth germ.