康复新治疗牙周炎的机制研究

Mechanism of Kangfuxin in treating periodontitis

目的通过研究康复新对牙周组织相关细胞增殖的影响及康复新在细胞炎症反应模型中的抗炎作用,从促进牙周组织再生和抗炎2个方面探讨康复新治疗牙周炎的相关机制。方法(1)采用淋巴细胞增殖检测法(MTS法)检测不同浓度康复新对人牙周膜成纤维细胞(hPDLFs)、人牙龈上皮细胞(hGECs)、人单核巨噬细胞(THP-1)和小鼠胚胎成骨细胞(MC3T3-E1)活性的影响;(2)通过细菌脂多糖(LPS)刺激小鼠巨噬细胞RAW264.7建立细胞炎症模型,采用实时定量聚合酶链反应(qRT-PCR)法检测康复新对细胞白细胞介素-1β(IL-1β)、IL-10、一氧化氮合酶(NOS)和基质金属蛋白酶-13(MMP-13)mRNA表达水平的影响。结果(1)与对照组比较:0.1000、0.0500、0.0250、0.0125 mg/mL的康复新组在24 h和48 h时间点均可刺激hPDLFs、THP-1和MC3T3-E1增殖(P<0.01),且促增殖作用具有浓度依赖性;(2)在炎症细胞模型中,与对照组比较,0.0500 mg/mL和0.0125 mg/mL的康复新组IL-1β和IL-10 mRNA表达水平比较,差异均无统计学意义(P>0.05),NOS和MMP-13 mRNA表达水平降低,差异有统计学意义(P<0.05)。结论康复新在一定浓度范围内能促进hPDLFs、hGECs、THP-1和MC3T3-E1增殖;康复新可抑制LPS诱导的RAW264.7细胞炎症反应,其抗炎机制可能是降低NOS和MMP-13 mRNA表达水平。康复新可能是通过促进牙周组织再生和抗感染治疗牙周炎。

Objective To study the effect of Kangfuxin on the proliferation of periodontal tissue-related cells and the anti-inflammatory effect of Kangfuxin in the cell inflammatory response model,and to explore the related mechanism of Kangfuxin in treating periodontitis from the aspects of promoting periodontal tissue regeneration and anti-inflammatory.Methods(1)MTS method was used to detect the effects of different concentrations of Kangfuxin on the activities of Human Periodontal Ligament Fibroblasts(hPDLFs),human Gingival Epithelial Cells(hGECs),THP-1 and MC3T3-E1 proliferation.(2)The cell inflammation model was established by stimulating mouse macrophage RAW264.7 with bacterial lipopolysaccharide(LPS).The quantitative Real-time polymerase chain reaction(qRT-PCR)was used to detect the effects of Kangfuxin on the mRNA expression of interleukin-1β(IL-1β),IL-10,nitric oxide synthase(NOS)and matrix metalloproteinase-13(MMP-13).Results(1)Compared with the control group:the 0.1000,0.0500,0.0250,0.0125 mg/mL rehabilitation groups could stimulate the proliferation of hPDLFs,THP-1 and MC3T3-E1 at 24 h and 48 h time points(P<0.01),and the proliferation promotion effect was concentration-dependent.(2)In the inflammatory cell model,compared with the control group,there was no significant differences in IL-1βand IL-10 mRNA expression levels in the 0.05 mg/mL group and the 0.0125 mg/mL group(P>0.05).The mRNA levels of NOS and MMP-13 we re decreased,and the difference was statistically significant(P<0.05).Conclusion Kangfuxin promote the proliferation of hPDLFs,hGECs,THP-1 and MC3T3-E1 in a certain concentration range.Kangfuxin inhibit LPS-induced inflammatory response of RAW264.7,and its anti-inflammatory mechanism may be the decrease of mRNA expression of NOS and MMP-13.Kangfuxin may treat periodontitis by promoting periodontal tissue regeneration and anti-inflammation.