State Key Laboratory of Military Stomatology and National Clinical Research Center for Oral Diseases and Shaanxi Key Laboratory of Stomatology, Department of Prosthodontics, School of Stomatology, Fourth Military Medical University, 145 West Changle Road, Xi'an, 710032, People's Republic of China.
Arch Oral Biol. 2017 Nov;83:111-117. doi: 10.1016/j.archoralbio.2017.07.006. Epub 2017 Jul 20.
This study was aimed to evaluate the anti-matrix metalloproteinases (MMPs) ability of active components from citrus fruits (hesperetin: Hst, hesperidin: Hsd and naringenin: Nge).
Inactivation effects of citrus flavonoids (Hst, Hsd, Nge) at different concentrations on soluble collagenase were measured using a fluorometric assay. Matrix-bound endogenous MMPs activity was evaluated via dry mass loss and hydroxyproline (HYP) release of demineralized human dentin. Demineralized dentin beams were pretreated with 500μg/mL citrus flavonoids for 10min. Chlorhexidine (CHX) was used as inhibitor control. Beams pretreated with distilled water served as blank control. Dentin slabs were used for in situ zymography and evaluated under confocal microscopy. Ultrastructure of demineralized collagen fibers was exhibited by Transmission Electron Microscopy (TEM).
Citrus flavonoids exhibited inactivation function on soluble MMPs and the extent of inactivation increased in a dose-dependent manner. The inactivation percent of citrus flavonoids reached above 90% at the concentration of 500μg/mL. Compared with control group, citrus flavonoids pretreated demineralized dentin beams exhibited less dry mass loss, lower hydroxyproline release and more intact collagen architecture after 15days storage. Dentin samples pretreated with citrus flavonoids showed lower enzymes activities in in situ zymography.
Hst, Hsd or Nge have anti-MMPs ability and can preserve dentin collagen from degradation.
Hst, Hsd and Nge may have the potential to be used in dentin bonding systems and improve the resin-dentin bonding durability.
本研究旨在评估柑橘类水果(桔皮素:Hst、桔皮苷:Hsd 和柚皮苷:Nge)中的活性成分对基质金属蛋白酶(MMPs)的抑制作用。
采用荧光法测定不同浓度柑橘类黄酮(Hst、Hsd、Nge)对可溶性胶原酶的失活作用。通过干燥失重和脱矿人牙本质羟脯氨酸(HYP)释放来评估基质结合内源性 MMPs 活性。用 500μg/mL 柑橘类黄酮预处理脱矿牙本质梁 10min,氯己定(CHX)作为抑制剂对照。蒸馏水预处理的牙本质梁作为空白对照。用原位酶谱法对牙本质薄片进行处理,并在共聚焦显微镜下进行评估。脱矿胶原纤维的超微结构通过透射电子显微镜(TEM)显示。
柑橘类黄酮对可溶性 MMPs 具有失活作用,失活程度随浓度的增加呈剂量依赖性增加。柑橘类黄酮的失活率在 500μg/mL 浓度下超过 90%。与对照组相比,经柑橘类黄酮预处理的脱矿牙本质梁在储存 15 天后表现出较低的干燥失重、羟脯氨酸释放量和更完整的胶原结构。原位酶谱法显示,用柑橘类黄酮预处理的牙本质样本中的酶活性较低。
Hst、Hsd 或 Nge 具有抗 MMPs 能力,可以防止牙本质胶原降解。
Hst、Hsd 和 Nge 可能具有用于牙本质粘结系统的潜力,并提高树脂-牙本质粘结的耐久性。