血竭提取物对牙龈炎大鼠牙周组织修复及TLR4/NF-κB信号通路的影响

Effect of dragon’s blood extract on periodontal tissue repair and TLR4/NF-κB signal pathway

目的:探讨血竭提取物对牙龈炎大鼠牙周组织修复及Toll样受体4/核转录因子kappa B(TLR4/NF-κB)通路的影响。方法:将50只大鼠随机分为对照组、牙龈炎组及血竭提取物低、中、高剂量组,每组10只。除对照组外,其余各组均采用丝线结扎法建立牙龈炎大鼠模型;模型建立成功后,血竭提取物低、中、高剂量组大鼠分别给予150、300、600 mg·kg^(-1)·d^(-1)血竭提取物灌胃处理(1次/d,连续4周),模型组、对照组大鼠同时间给予等量生理盐水灌胃处理。麻醉处死大鼠,取左侧上颌第二磨牙颌骨组织,以亚甲基蓝染色观察并测定牙槽骨丧失(loss of alveolar bone,ABL)情况;H-E染色观察颌骨组织病理学改变;酶联免疫吸附试剂盒(ELISA)法检测各组大鼠牙周组织(颌骨组织)IL-17、IL-4水平;免疫印迹法检测大鼠牙周组织(颌骨组织)骨形成蛋白2(bone morphogenetic protein-2,BMP-2)及TLR4、NF-κB p65蛋白水平。采用SPSS 19.0软件包对数据进行统计学分析。结果:与对照组相比,模型组大鼠ABL、颌骨组织中IL-17、IL-4及TLR4、NF-κB p65蛋白水平显著升高(P<0.05),颌骨组织中BMP-2蛋白水平显著降低(P<0.05);与模型组相比,血竭提取物低、中、高剂量组大鼠ABL、颌骨组织中IL-17、IL-4及TLR4、NF-κB p65蛋白水平显著降低(P<0.05),颌骨组织中BMP-2蛋白水平显著升高(P<0.05),呈剂量依赖性。结论:血竭提取物可能通过抑制TLR4/NF-κB通路活化,抑制炎症反应,促进牙龈炎大鼠牙周组织修复。

PURPOSE:To discuss the effects of dragon’s blood extract,a Chinese herb,on periodontal tissue repair and toll like receptor 4/nuclear transcription factor kappa B(TLR4/NF-κB) in gingivitis rats.METHODS:Sixty rats were randomly divided into control group,gingivitis group and low,medium and high dose groups of dragon’s blood extract,with 10rats in each group.Except the control group,the gingivitis rat model was established by silk thread ligation in other groups.The model was established successfully.The low,medium and high dose groups rats were given 150,300 and 600mg·kg^(-1)·d^(-1)dragon’s blood extract successively by gavage(once a day for 4 weeks).Rats in the model group and the control group were given the same amount of normal saline by gavage at the same time.After the rats were sacrificed under anesthesia,the jaw tissue of the left maxillary second molar was stained with methylene blue to observe and measure the loss of alveolar bone(ABL),H-E staining was used to observe the pathological changes of periodontal tissue(jaw tissue).The levels of IL-17 and IL-4 in periodontal tissue(jaw tissue) of rats in each group were detected by enzyme-linked im-munosorbent assay(ELISA).Bone morphogenetic protein-2(BMP-2),TLR4 and NF-κB p65 protein level in rat periodontal tissue were detected by Western blot.SPSS 19.0 software package was used to analyze the data.RESULTS:Compared with the control group,IL-17,IL-4,TLR4 and NF-κB p65 protein and ABL in jaw tissue of model group were significantly increased(P<0.05),and the level of BMP-2 protein in jaw tissue significantly decreased(P<0.05).Compared with the model group,IL-17,IL-4,TLR4 and NF-κB p65 protein and ABL in jaw tissue of rats in low,medium and high dose groups of dragon’s blood extract were significantly increased,and the level of BMP-2 protein significantly decreased(P<0.05).CONCLUSIONS:Dragon’s blood extract can inhibit TLR4/NF-κB.Activation of B pathway inhibits inflammatory response and promotes periodontal tissue repair in gingivitis rats.