黄芩苷抑制炎性因子对牙周炎症的影响及其相关机制研究

Study on Effect of Baicalin on Periodontitis by Inhibiting Inflammatory Factors and Its Related Mechanism

目的:探讨黄芩苷对牙周炎症的影响,并研究其相关分子机制。方法:将50只无特定病原级雌性大鼠分为空白组(不作任何处理)、对照1组(于右侧上颌第2磨牙颊侧正中龈沟底注射生理盐水)、对照2组[于右侧上颌第2磨牙颊侧正中龈沟底注射含脂多糖(LPS)的生理盐水]、实验1组(于右侧上颌第2磨牙颊侧正中龈沟底注射含LPS的生理盐水和浓度为0.1μg/mL的黄芩苷)、实验2组(于右侧上颌第2磨牙颊侧正中龈沟底注射含LPS的生理盐水和浓度为1.0μg/mL的黄芩苷),每组10只。实验第9天处死一部分大鼠取上颌骨,Micro-CT扫描检测牙槽骨吸收情况;另一部分大鼠取其静脉血培养巨噬细胞,逆转录-聚合酶链式反应(RT-PCR)法检测转化生长因子-β(TGF-β)m RNA表达,Western blot法检测TGF-β、白细胞介素-1β(IL-1β)、胱天蛋白酶-1(Caspase-1)、高迁移率族蛋白B1(HMGB1)表达。结果:对照1组各检测指标与空白组比较,差异均无统计学意义(P>0.05)。与对照1组比较,对照2组大鼠骨体积、骨表面体积比、骨小梁厚度、骨小梁数目、巨噬细胞TGF-β蛋白及mRNA、IL-1β、Caspase-1、HMGB1表达均下降(P<0.05),而骨小梁模型因子及骨小梁间隙均上升(P<0.05)。与对照2组比较,实验1组、实验2组大鼠骨体积、骨表面体积比、骨小梁厚度、骨小梁数目、巨噬细胞TGF-β蛋白及mRNA、IL-1β、Caspase-1、HMGB1表达均增加(P<0.05),骨小梁模型因子、骨小梁间隙均减小(P<0.05)。与实验1组比较,实验2组大鼠骨体积、骨表面体积比、骨小梁厚度、骨小梁数目、巨噬细胞TGF-β蛋白及mRNA、IL-1β、Caspase-1、HMGB1表达均增加(P<0.05);骨小梁模型因子、骨小梁间隙均减小(P<0.05)。结论:黄芩苷可促进巨噬细胞分泌TGF-β,激活Caspase-1/IL-1β通路,促进诱导的破骨细胞形成减少,从而减少牙槽骨破坏和吸收,减轻牙周炎症。

Objective:To discuss the effect of baicalin on periodontitis and study its related molecular mechanism.Methods:A total of 50 specific pathogen free-grade female rats were divided into the blank group without any treatment,the Control Group One(injection of normal saline into the bottom of median gingival sulcus of maxillary second molar on the right side of buccal surface),the Control Group Two(injection of normal saline containing lipopolysaccharide(LPS)into the same location as that of the Control Group One),the Experiment Group One(injection of normal saline containing LPS and0.1μg/mL baicalin into the same location as that of the Control Group One),and the Experiment Group 2(injection of normal saline containing LPS and 1.0μg/mL baicalin into the same location as that of the Control Group One),10 rats in each group.On the 9 th day of experiment,a part of rats were sacrificed,and their maxillary bone was taken out;alveolar bone resorption was examined by Micro-CT.On the 9 th day of experiment,venous blood was taken from the rest part of rats to culture macrophage;the expression of m RNA of transforming growth factor-β(TGF-β)was detected by reverse transcriptase polymerase chain reaction(RT-PCR);the protein expression of TGF-βand Interleukin-1β(IL-1β),Caspase-1,high mobility group box 1(HMGB1)was detected by Western blot.Results:There were no significant differences being found in the comparison of each detection index between the Control Group One and the blank group(P>0.05).Compared with those in the Control Group One,in the Control Group Two,bone volume,the ratio of bone surface to bone volume,the thickness and number of bone trabecula,and the expression of protein and m RNA of macrophage like TGF-β1,as well as the expression of IL-1β,caspase-1,and HMGB1 marked by macrophage were decreased(P<0.05);pattern factor and spacing of bone trabecula were increased(P<0.05).Compared with those in the Control Group Two,in the Experiment Group One and Group Two,bone volume,the ratio of bone surface to bone volume,the thickness and number of bone trabecula,and the expression of protein and mRNA of macrophage like TGF-β1,as well as the expression of IL-1β,Caspase-1,and HMGB1 were increased(P<0.05);pattern factor and spacing of bone trabecula were decreased(P<0.05).Compared with those in the Experiment Group One,in the Experiment Group 2,bone volume,the ratio of bone surface to bone volume,the thickness and number of bone trabecula,and the expression of protein and mRNA of macrophage like TGF-β1,as well as the expression of IL-1β,Caspase-1,and HMGB1 were increased(P<0.05);pattern factor and spacing of bone trabecula were decreased(P<0.05).Conclusion:Baicalin can promote secretion of TGF-βby macrophage and reduction of induced osteoclastogenesis,as well as activate the Caspase-1/IL-1βpathway,thus reducing alveolar bone destruction and resorption,as well as relieving periodontitis.