中华口腔医学杂志
中華口腔醫學雜誌
중화구강의학잡지
Chinese Journal of Stomatology
2013年
12期
734-739
,共6页
靳趁心%乔治向%刘冰%陈东来%王永兰
靳趁心%喬治嚮%劉冰%陳東來%王永蘭
근진심%교치향%류빙%진동래%왕영란
β防御素%前列腺素E类%基质金属蛋白酶1%牙龈成纤维细胞
β防禦素%前列腺素E類%基質金屬蛋白酶1%牙齦成纖維細胞
β방어소%전렬선소E류%기질금속단백매1%아간성섬유세포
Beta-defensins%Prostaglandins E%Matrix metalloproteinase 1%Gingival fibroblasts
目的 观察人类β防御素3(human β-defensin-3,HBD-3)对人牙龈成纤维细胞(human gingival fibroblasts,HGF)增殖及分泌前列腺素E2(prostaglandin E2,PGE2)、基质金属蛋白酶1(matrix metalloproteinase-1,MMP-1)的影响,了解HBD-3调节牙周免疫的分子机制.方法 采用组织块原代培养法培养HGF,取P4代细胞接种于96孔板,分为对照组和4个实验组,实验组分别加入100μl质量浓度为0.3、1.0、3.0、10.0 mg/L的HBD-3,对照组加入等量达尔伯克改良伊格尔培养基(Dulbecco's modified Eagle's medium,DMEM)培养液,37 ℃、5％ C02、饱和湿度条件下培养7d,以甲基噻唑基四唑法检测各组HGF的增殖情况并绘制生长曲线,以酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)法检测各组12h时培养上清液中PGE2及MMP-1的含量.各组结果数据的总体比较采用单因素方差分析,组间比较采用双侧t检验,以P＜0.05为差异有统计学意义.结果 细胞生长曲线显示各组HGF数量均随时间延长而增加,且与HBD-3质量浓度呈一定的依赖性;ELISA结果显示1.0 mg/L HBD-3组培养上清液中PGE2及MMP-1含量最高,分别为(350.56 ±63.96) ng,/L和(13.22± 0.59) μg/L,且与对照组、10.0 mg/L HBD-3组相比差异均有统计学意义(P＜0.05).结论 HBD-3可以促进HGF增殖,且呈浓度依赖性;低质量浓度的HBD-3上调HGF的PGE2及MMP-1分泌,在牙周免疫反应中发挥一定作用.
目的 觀察人類β防禦素3(human β-defensin-3,HBD-3)對人牙齦成纖維細胞(human gingival fibroblasts,HGF)增殖及分泌前列腺素E2(prostaglandin E2,PGE2)、基質金屬蛋白酶1(matrix metalloproteinase-1,MMP-1)的影響,瞭解HBD-3調節牙週免疫的分子機製.方法 採用組織塊原代培養法培養HGF,取P4代細胞接種于96孔闆,分為對照組和4箇實驗組,實驗組分彆加入100μl質量濃度為0.3、1.0、3.0、10.0 mg/L的HBD-3,對照組加入等量達爾伯剋改良伊格爾培養基(Dulbecco's modified Eagle's medium,DMEM)培養液,37 ℃、5％ C02、飽和濕度條件下培養7d,以甲基噻唑基四唑法檢測各組HGF的增殖情況併繪製生長麯線,以酶聯免疫吸附測定(enzyme-linked immunosorbent assay,ELISA)法檢測各組12h時培養上清液中PGE2及MMP-1的含量.各組結果數據的總體比較採用單因素方差分析,組間比較採用雙側t檢驗,以P＜0.05為差異有統計學意義.結果 細胞生長麯線顯示各組HGF數量均隨時間延長而增加,且與HBD-3質量濃度呈一定的依賴性;ELISA結果顯示1.0 mg/L HBD-3組培養上清液中PGE2及MMP-1含量最高,分彆為(350.56 ±63.96) ng,/L和(13.22± 0.59) μg/L,且與對照組、10.0 mg/L HBD-3組相比差異均有統計學意義(P＜0.05).結論 HBD-3可以促進HGF增殖,且呈濃度依賴性;低質量濃度的HBD-3上調HGF的PGE2及MMP-1分泌,在牙週免疫反應中髮揮一定作用.
목적 관찰인류β방어소3(human β-defensin-3,HBD-3)대인아간성섬유세포(human gingival fibroblasts,HGF)증식급분비전렬선소E2(prostaglandin E2,PGE2)、기질금속단백매1(matrix metalloproteinase-1,MMP-1)적영향,료해HBD-3조절아주면역적분자궤제.방법 채용조직괴원대배양법배양HGF,취P4대세포접충우96공판,분위대조조화4개실험조,실험조분별가입100μl질량농도위0.3、1.0、3.0、10.0 mg/L적HBD-3,대조조가입등량체이백극개량이격이배양기(Dulbecco's modified Eagle's medium,DMEM)배양액,37 ℃、5％ C02、포화습도조건하배양7d,이갑기새서기사서법검측각조HGF적증식정황병회제생장곡선,이매련면역흡부측정(enzyme-linked immunosorbent assay,ELISA)법검측각조12h시배양상청액중PGE2급MMP-1적함량.각조결과수거적총체비교채용단인소방차분석,조간비교채용쌍측t검험,이P＜0.05위차이유통계학의의.결과 세포생장곡선현시각조HGF수량균수시간연장이증가,차여HBD-3질량농도정일정적의뢰성;ELISA결과현시1.0 mg/L HBD-3조배양상청액중PGE2급MMP-1함량최고,분별위(350.56 ±63.96) ng,/L화(13.22± 0.59) μg/L,차여대조조、10.0 mg/L HBD-3조상비차이균유통계학의의(P＜0.05).결론 HBD-3가이촉진HGF증식,차정농도의뢰성;저질량농도적HBD-3상조HGF적PGE2급MMP-1분비,재아주면역반응중발휘일정작용.
Objective To observe the effect of human β-defensin-3 (HBD-3) on proliferation and the secretion of prostaglandin E2(PGE2) and matrix metalloproteinase-1 (MMP-1) in human gingival fibroblasts(HGF).Methods The HGF were cultured with tissue-explant method and the fourth-generation HGF were plated in 96-well plate.All groups except the control group were treated with different concentrations of HBD-3 for 7 days.Then the HGF proliferation was evaluated with methyl thiazolyl tetrazolium(MTT) colorimetry and the secretions of PGE2 and MMP-1 at the 12th hours of each group were detected by enzyme-linked immunosorbent assay(ELISA).Results The result of MTT dynamic monitoring showed that the amount of HGF increased with time in all groups in concentration dependent manner.ELISA showed that the secretions of PGE2 and MMP-1 in 1.0 mg/L HBD-3 group were (350.56 ± 63.96) ng/L and (13.22 ±0.59) μg/L,significantly higher than those in the control group and 10.0 mg/L HBD-3 group (P ＜ 0.05).Conclusions HBD-3 promoted the proliferation of HGF.The low concentration of HBD-3 may play a role in immunoregulation through increasing the secretions of PGE2 and MMP-1.
