Effects of periodontitis patient’s own tissue nucleic acid on the mRNA expression of osteoclast-related factors in murine macrophages
10.7518/hxkq.2015.02.018
- VernacularTitle:牙周炎患者自身组织核酸刺激小鼠巨噬细胞后对破骨相关因子mRNA表达的影响
- Author:
Ziqing DING
1
;
Yuqin SHEN
;
Yue ZHOU
;
Yin LIU
;
Han GAO
;
Haijiao YU
;
Chongtao LIN
Author Information
1. 吉林大学口腔医院牙周病科
- Keywords:
specific sequence oligodeoxynucleotide MT01;
periodontitis patient’s own tissue nucleic acid;
osteoclast-related factors;
murine macrophages
- From:
West China Journal of Stomatology
2015;(2):192-196
- CountryChina
- Language:Chinese
-
Abstract:
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.
