CAJ | 학술논문

目的：检测牙周炎患者自身组织核酸刺激巨噬细胞后破骨相关因子白细胞介素-6（IL-6）、白细胞介素-12（IL-12）p35、IL-12p40、基质金属蛋白酶-9（MMP-9）、活化T细胞核因子?1（NFATc1）、核激活因子κB受体（RANK）、肿瘤坏死因子-α（TNF-α）mRNA的表达，观察牙周炎患者自身组织核酸对巨噬细胞向破骨细胞分化的影响作用。方法采集翻瓣术中慢性牙周炎患者炎症牙周组织及正畸患者健康牙拔除术获取的健康牙周组织，提取组织总RNA逆转录cDNA。培养小鼠巨噬细胞系RAW264.7，加入质量浓度1?μg·mL-1的特定序列寡核苷酸MT01共孵育3?h后（以1?μg·mL-1的PBS作为对照），加入已提取的炎症牙周组织及健康牙周组织cDNA（质量浓度为1?μg·mL-1）。实验分4组：健康组织cDNA，炎症组织cDNA，MT01+健康组织cDNA，MT01+炎症组织cDNA。4组细胞分别孵育3、6、12、24?h，采用实时定量聚合酶链反应法检测破骨相关因子IL-6、IL-12p35、IL-12p40、MMP-9、NFATc1、RANK及TNF-α mRNA的表达，进行两两组间比较。结果牙周炎患者自身组织核酸可上调RAW264.7破骨相关因子IL-6、IL-12p35、IL-12p40、MMP-9、NFATc1、RANK及TNF-α mRNA的表达；在免疫抑制剂MT01的作用下，牙周炎患者自身组织核酸上调RAW264.7内破骨相关因子mRNA的表达状况受到抑制。结论牙周炎患者自身组织核酸可以影响小鼠巨噬细胞向破骨细胞的分化。
목적：검측아주염환자자신조직핵산자격거서세포후파골상관인자백세포개소-6（IL-6）、백세포개소-12（IL-12）p35、IL-12p40、기질금속단백매-9（MMP-9）、활화T세포핵인자?1（NFATc1）、핵격활인자κB수체（RANK）、종류배사인자-α（TNF-α）mRNA적표체，관찰아주염환자자신조직핵산대거서세포향파골세포분화적영향작용。방법채집번판술중만성아주염환자염증아주조직급정기환자건강아발제술획취적건강아주조직，제취조직총RNA역전록cDNA。배양소서거서세포계RAW264.7，가입질량농도1?μg·mL-1적특정서렬과핵감산MT01공부육3?h후（이1?μg·mL-1적PBS작위대조），가입이제취적염증아주조직급건강아주조직cDNA（질량농도위1?μg·mL-1）。실험분4조：건강조직cDNA，염증조직cDNA，MT01+건강조직cDNA，MT01+염증조직cDNA。4조세포분별부육3、6、12、24?h，채용실시정량취합매련반응법검측파골상관인자IL-6、IL-12p35、IL-12p40、MMP-9、NFATc1、RANK급TNF-α mRNA적표체，진행량량조간비교。결과아주염환자자신조직핵산가상조RAW264.7파골상관인자IL-6、IL-12p35、IL-12p40、MMP-9、NFATc1、RANK급TNF-α mRNA적표체；재면역억제제MT01적작용하，아주염환자자신조직핵산상조RAW264.7내파골상관인자mRNA적표체상황수도억제。결론아주염환자자신조직핵산가이영향소서거서세포향파골세포적분화。
ObjectiveThis?paper?aimed?to?determine?the?mRNA?expression?of?osteoclast-related?factors?interleukin-6?(IL-6),?interleukin-12?(IL-12)?p35,?IL-12p40,?matrix?metalloproteinase-9?(MMP-9),?nuclear?factor?of?activated?T-cells?cyto-plasmic 1 (NFATc1), receptor activator of nuclear factor-κB (RANK), and tumor necrosis factor-α (TNF-α) mRNA in murine macrophages?infected?by?a?periodontitis?patient’s?own?tissue?nucleic?acid.?Another?aim?was?to?investigate?the?effects?of?a?perio-dontitis?patient’s?own?tissue?nucleic?acid?on?the?differentiation?of?macrophages?into?osteoclasts.?Methods???Inflammatory?periodontal?tissue?samples?of?chronic?periodontitis?patients?were?taken?during?periodontal?flap?surgery,?and?healthy?gingival?tissue?samples?were?taken?from?orthodontic?patients?during?tooth?extractions.?Total?RNA?from?periodontal?tissue?was?extracted?and?reversely?transcribed?into?cDNA?and?then?cryo-preserved?until?further?use.?First,?specific?sequence?oligodeoxynucleotide?MT01?at?a?concentration?of?1?μg·mL-1?was?added?in?murine?macrophage?RAW264.7,?and?the?cells?were?incubated?for?3?hours.?Cells?with?PBS?(1?μg·mL-1)?were?used?as?negative?controls.?The?inflammatory?periodontal?tissue?cDNA?and?healthy?periodontal?tissue?cDNA?(1?μg·mL-1)?was?added?subsequently.?There?were?four?experimental?groups:?healthy?periodontal?tissue?cDNA+RAW264.7,?inflammatory?periodontal?tissue?cDNA+RAW264.7,?MT01+healthy?periodontal?tissue?cDNA+RAW264.7,?and?MT01+inflammatory?periodontal?tissue?cDNA+RAW264.7.?Real-time?quantitative?polymerase?chain?reaction?was?used?to?detect?the?mRNA?expression?of?osteoclast-related?factors?IL-6,?IL-12p35,?IL-12p40,?MMP-9,?NFATc1,?RANK,?and?TNF-α mRNA after 3, 6, 12, and 24 hours.Results???The?mRNA?levels?of?osteoclast-related?factors?NFATc1,?MMP-9,?TNF-α, IL-6, IL-12p40, IL-12p35, and RANK in RAW264.7 were markedly upregulated?with?the?treatment?of?periodontitis?patient’s?own?tissue?nucleic?acid.?However,?the?mRNA?expression?of?osteoclast-related?factors?was?inhibited?by?use?of?an?im-munosuppressant?MT01.?Conclusion???The?periodontitis?patient’s?own?tissue?nucleic?acid?could?promote?the?differentiation?of?murine?macrophage?into?osteoclasts.