Analysis of gene expression concerned with PDL mechanical receptor in biting

咬合过程中PDL机械受体相关基因表达分析

• 批准号: 12671896
• 负责人: BABA Shunsuke
• 金额: $ 1.73万
• 依托单位: Kyoto Prefectural University of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671896/
• 关键词: cytokine; PDL; 咬合接触; 咬合挙上

Periodontal ligament (PDL) cells, which consist of osteoblasts, fibloblasts, osetoclasts, and et.al., maintain the attachment of the tooth to alveolar bone. Recently, it has been reported that PDL cells examines function and characterization in vitro. To confirm immune response for periodontal disease casing dental plaque on PDL cells, we investigated the cytokine Mrna expression and production stimulating Prevotella intermedia and Streptococcus mutans. PDL cells were obtained from the middle third of the root surface of healthy human erupted maxillary third molar. The tissue minced and cultured as explants in tissue culture medium [D-MEM with D-Glucose 4500 mg/dl L-Glu NaHCO3 ] containing 10% low-endotoxin fetal calf serum (FCS), 100U penisillin, 25ug/ml streptomysin, amphotericin B as FugizoneTM in 0.85% saline). Cultures were then grown in medium and cloned by limiting dilutions, I.e., the cells obtained from primary cultures. The confluent cultures were used for experimentation bet … More ween the 3th and 4th passages. Bacteria were the kind gift from department of Microbiology, Kyoto Prefectural University of Medicine. Cultures were confluent (1.0× 105cells/well). And stimulated by bacteria (1.0×107/ml) after 24 hours in a 6-well plate. No bacteria were used as control. Total RNA was extracted by ISOGENTM (NIPPON GENE). 1mg of RNA was used for Cdna synthesis with a reverse transcriptase (SuperscriptII RNAse H; GIBCO-BRL) and an oligo (dt) 16 primer. Reaction mixture was used as a template for PCR amplification was run for 35 cycles at 94℃ for 1 min, 60℃ for 1 min, and 72℃ for 1 min. PCR product was analyzed by electrophoresis on a 1.5% agarose gel containing ethidium bromide, and the bands were examined under UV light. The expression of IL-1b, IL-6, IL-8, TNF-α, TGF-α, and TGF-β were detected. This study indicated expression of many inflammatory cytokines. These results suggest that there was a relationship between the cytokine expression and the inflammation of periodontal ligaments on pockets. Furthermore, we may examine immune mechanism which marginal periodontitis affect mechanical stress and Bone Morphogenetic Protein Less

牙周膜 (PDL) 细胞由成骨细胞、成纤维细胞、破骨细胞等组成，维持牙齿与牙槽骨的附着。最近，有报道称 PDL 细胞可在体外检查功能和表征。为了确认 PDL 细胞上牙周病牙菌斑的免疫反应，我们研究了刺激中间普雷沃氏菌和变形链球菌的细胞因子 Mrna 表达和产生。 PDL细胞取自健康人已萌出的上颌第三磨牙根面的中间三分之一处。将组织切碎并作为外植体在组织培养基[D-MEM与D-葡萄糖4500 mg/dl L-Glu NaHCO3]中培养，该培养基含有10%低内毒素胎牛血清(FCS)、100U青霉素、25ug/ml链霉素、两性霉素B（作为FugizoneTM的0.85%盐水）。然后将培养物在培养基中生长并通过有限稀释进行克隆，即从原代培养物中获得的细胞。汇合的培养物用于第三和第四代的实验。细菌是京都府立医科大学微生物学系赠送的礼物。培养物汇合（1.0×105细胞/孔）。并在6孔板中用细菌（1.0×107/ml）刺激24小时后。不使用细菌作为对照。通过ISOGENTM(NIPPON GENE)提取总RNA。使用逆转录酶（SuperscriptII RNAse H；GIBCO-BRL）和寡核苷酸（dt）16 引物将 1mg RNA 用于 Cdna 合成。以反应混合物为模板进行PCR扩增，94℃1分钟、60℃1分钟、72℃1分钟运行35个循环。 PCR产物在含有溴化乙锭的1.5%琼脂糖凝胶上进行电泳分析，并在紫外光下检查条带。检测IL-1b、IL-6、IL-8、TNF-α、TGF-α、TGF-β的表达。这项研究表明许多炎症细胞因子的表达。这些结果表明细胞因子的表达与牙袋上牙周膜的炎症之间存在关系。此外，我们还可以检查边缘性牙周炎影响机械应力和骨形态发生蛋白的免疫机制。

项目成果

Namba M, Yamada K, Kudeken W, Mizutani G, Tsukitani K, Mori M, Takai Y: "Phenotypic changes of calponin in salivary gland myoepithelial cells during postnatal development of rats"Acta Histochem, Cytochem. 33(2). 73-79 (2000)

Namba M、Yamada K、Kudeken W、Mizutani G、Tsukitani K、Mori M、Takai Y：“大鼠出生后发育过程中唾液腺肌上皮细胞中钙调蛋白的表型变化”Acta Histochem，Cytochem。

Masataka Itoigawa, Chihiro Ito, Hugh T.W.Tan, Masato Okuda, Harukuni Tokuda, Hoyoku Nishino, Hiroshi Furukawa: "Cancer chemoprevetive activity of naphthoquinones and their analogs From Avicennia plants"Cancer Letters. 174・2. 135-139 (2001)

Masataka Itoikawa、Chihiro Ito、Hugh T.W.Tan、Masato Okuda、Harukuni Tokuda、Hoyoku Nishino、Hiroshi Furukawa：“来自白骨壤植物的萘醌及其类似物的癌症化学预防活性”癌症快报 174・2。

Yamada K, Namba M, Sumitomo S, Mori M, Tsukitani K, Shrestha P, Takai Y: "Heterogeneity of expression of calponin and metallothionein in reactive myoepithehal/ductal basal cells of obstructive sialadenitis"Acta Histochem.Cytochem.. 33・4. 287-294 (2000)

山田K、难波M、住友S、森M、月谷K、Shrestha P、高井Y：“阻塞性唾液腺炎反应性肌上皮/导管基底细胞中钙调蛋白和金属硫蛋白表达的异质性”组织化学学报.细胞化学.. 33・4。 287-294 (2000)

Namba M, Yamada K, Kudeken W, Mizutani G, Tsukikani K, Mori M, Takai Y: "Phenotypic changes of calponin in salivary gland myoepithelial cells during postnatal development of rats"Acta Histochem. Cytochem.. 33・2. 73-79 (2000)

Namba M、Yamada K、Kudeken W、Mizutani G、Tsukikani K、Mori M、Takai Y：“大鼠出生后发育过程中唾液腺肌上皮细胞中钙调蛋白的表型变化”细胞化学学报.. 73・2。 (2000)

Namba M, Yamada K, Kudeken W, Mizutani G, Tsukitani K, Mori M, Takai Y: "Phenotypic changes of calponin in salivary gland myoepithelial cells during postnatal development of rats"Acta Histochem.Cytochem.. 33・2. 73-79 (2000)

Namba M、Yamada K、Kudeken W、Mizutani G、Tsukitan K、Mori M、Takai Y：“大鼠出生后发育过程中唾液腺肌上皮细胞中钙调蛋白的表型变化”Acta Histochem.Cytochem.. 73・2。 (2000)