夏枯草黄酮抑制大鼠牙周炎低氧诱导因子1α表达的治疗机制
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摘要
目的:了解夏枯草黄酮治疗大鼠牙周炎过程中所靶向抑制的低氧诱导因子1α(hypoxia-inducible factor-1α,HIF-1α)的表达情况。方法:取40只无特定病原体(specefic pathogen free,SPF)级雄性Wistar大鼠,按照随机数表法将其随机分为5组(8只/组),依次为正常对照组、模型组、夏枯草黄酮组(50、100、200mg/L),采用牙龈卟啉单胞菌(P.gingivalis)浸渍丝线结扎的方法构建牙周炎模型。连续处理8周,其中模型组与对照组替代药品选用等量生理盐水。利用光镜检查大鼠牙周炎大体标本,采用四级法测量牙龈指数,应用牙周探针探测牙周附着丧失的情况。牙龈组织与血清中HIF-1α表达情况采用酶联免疫吸附测定法检测。软骨细胞中HIF-1α表达水平采用免疫组化染色法检测。结果:模型组牙周炎症状典型,可见牙龈萎缩,牙周附着严重丧失,根部分叉可见暴露,且多可见累及两侧邻牙。给予夏枯草黄酮治疗后可见牙龈水肿逐步减轻,牙周袋逐步变浅,且牙龈萎缩与牙周附着丧失亦见好转。相比于正常对照组,模型组的大鼠牙龈指数、牙周附着丧失程度及HIF-1α表达量均可见增加(P<0.01)。相比于模型组,夏枯草黄酮组随着给药浓度的增加可见大鼠牙龈指数、牙周附着丧失程度及HIF-1α均呈逐渐下降的趋势(P<0.05)。相比于正常对照组中仅少数细胞HIF-1α呈弱阳性表达,模型组HIF-1α(+)细胞明显增加。但夏枯草黄酮各剂量组HIF-1α(+)细胞均见减少,且各剂量组HIF-1α(+)细胞率明显下降(P<0.01)。结论:夏枯草黄酮或主要通过降低HIF-1α的表达而实现其缓解牙周炎病情的目的。
Objective:To investigate the expression of hypoxia-inducible factor-1α(HIF-1α)in the treatment of rat periodontitis.Methods:Forty male Wistar rats with specific pathogen free(SPF)were randomly divided into 5 groups(n=8)according to random number table,i.e.normal control group,model group,and prunella vulgaris flavonoid groups(50,100,and 200 mg/L).P.gingivalis impregnated silk ligation was used to construct a periodontitis model.The treatment continued for 8 weeks,during which the model group and the control group were treated with the same amount of normal saline.Gross specimens of rat periodontitis were examined by light microscopy.The gingival index was measured by L9 e and Silness method,and the loss of periodontal attachment was detected by periodontal probe.The expression of HIF-1αin gingival tissues and serum was detected by enzyme-linked immunosorbent assay.The expression level of HIF-1αin chondrocytes was detected by immunohistochemical staining.Results:In the model group,periodontal inflammation was typical,and the gums were atro-phied.The periodontal attachment was severely lost.The furcation was exposed and more visible.After treatment with prunella vulgarisflavonoids,the gingival edema gradually reduced,the periodontal pockets gradually became shallower,and the gingival recession and periodontal attachment loss also improved.Compared with the normal control group,the gingival in-dex,degree of periodontal at tachment loss and HIF-1αexpression in the modelgroup all increased(P<0.01).Compared with the model group,the gingival index,periodontal attachment loss and HIF-1αof the prunella vulgaris flavonoid group showed a gradual decline with the increasing of the concentration(P<0.05).Compared with only a few cells in the normal control group,HIF-1αwas weakly positively expressed,and the HIF-1α(+)cells in the model group significantly increased.However,the HIF-1α(+)cells in the various doses of prunella vulgaris decreased,and the HIF-1α(+)cell rate significantly decreased in each dose group(P<0.01).Conclusion:Prunella vulgaris flavonoids can relieve periodontitis by reducing the expression of HIF-1α.
Objective:To investigate the expression of hypoxia-inducible factor-1α(HIF-1α)in the treatment of rat periodontitis.Methods:Forty male Wistar rats with specific pathogen free(SPF)were randomly divided into 5 groups(n=8)according to random number table,i.e.normal control group,model group,and prunella vulgaris flavonoid groups(50,100,and 200 mg/L).P.gingivalis impregnated silk ligation was used to construct a periodontitis model.The treatment continued for 8 weeks,during which the model group and the control group were treated with the same amount of normal saline.Gross specimens of rat periodontitis were examined by light microscopy.The gingival index was measured by L9 e and Silness method,and the loss of periodontal attachment was detected by periodontal probe.The expression of HIF-1αin gingival tissues and serum was detected by enzyme-linked immunosorbent assay.The expression level of HIF-1αin chondrocytes was detected by immunohistochemical staining.Results:In the model group,periodontal inflammation was typical,and the gums were atro-phied.The periodontal attachment was severely lost.The furcation was exposed and more visible.After treatment with prunella vulgarisflavonoids,the gingival edema gradually reduced,the periodontal pockets gradually became shallower,and the gingival recession and periodontal attachment loss also improved.Compared with the normal control group,the gingival in-dex,degree of periodontal at tachment loss and HIF-1αexpression in the modelgroup all increased(P<0.01).Compared with the model group,the gingival index,periodontal attachment loss and HIF-1αof the prunella vulgaris flavonoid group showed a gradual decline with the increasing of the concentration(P<0.05).Compared with only a few cells in the normal control group,HIF-1αwas weakly positively expressed,and the HIF-1α(+)cells in the model group significantly increased.However,the HIF-1α(+)cells in the various doses of prunella vulgaris decreased,and the HIF-1α(+)cell rate significantly decreased in each dose group(P<0.01).Conclusion:Prunella vulgaris flavonoids can relieve periodontitis by reducing the expression of HIF-1α.
引文
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[13]王斌,王小琴.慢性间歇低氧对大鼠牙周炎模型血清及牙龈组织中白细胞介素-18和低氧诱导因子-1α的影响[J].华西口腔医学杂志,2015,33(4):383-387.
[2] Himoto T,Fujita K,Nomura T,et al.Roles of copper in hepatocarcinogenesis via the activation of hypoxia-inducible factor-1α[J].Biol Trace Elem Res,2016,174(1):58-64.
[3] Zhang L,Zhang Y,Zhang X,et al.MicroRNA-433inhibits the proliferation and migration of HUVECs and neurons by targeting hypoxia-inducible factor 1alpha[J].J Mol Neurosci,2017,61(2):135-143.
[4]唐彩金,曾启新,方梅飞,等.消退素D1治疗实验性大鼠牙周炎的研究[J].口腔医学研究,2017,33(12):1270-1273.
[5]王2),顾思成,郭剑芬,等.高压氧疗法对大鼠心理应激相关牙周炎治疗作用及低氧诱导因子1α表达的研究[J].中国病理生理杂志,2016,32(2):339-346.
[6] Ishizuka S,Sakai T,Hiraiwa H,et al.Hypoxia-inducible factor-2αinduces expression of type X collagen and matrix metalloproteinases 13in osteoarthritic meniscal cells[J].Inflamm Res,2016,65(6):439-448.
[7] Ni JY,Xu LF,Wang WD,et al.Transarterial embolization combined with RNA interference targeting hypoxia-inducible factor-1αfor hepatocellular carcinoma:apreliminary study of rat model[J].J Cancer Res Clin Oncol,2017,143(2):199-207.
[8] Lee SY,Mustafa S,Ching YW,et al.Zinc induces normoxic accumulation of transcriptionally active hypoxia-inducible factor 1-alpha in mammary epithelial cells[J].Mol Biol(Mosk),2017,51(1):89-95.
[9]赵秀梅.中药夏枯草的药理作用研究现状[J].内蒙古中医药,2016,35(12):120-122.
[10] Li YJ,Zhang WB,Li SJ,et al.Prognosis value of Hypoxiainducible factor-1αexpression in patients with bone and soft tissue sarcoma:a meta-analysis[J].Springerplus,2016,5(1):1370.
[11]许丽华,徐莉,李彬,等.β-防御素2在实验性大鼠牙周炎口腔龈上皮、颊黏膜上皮及腮腺导管上皮的表达[J].口腔医学研究,2012,28(12):1217-1220.
[12]黄志刚,王耀,马克.低氧诱导因子1α基因在大鼠终板软骨细胞中的表达及意义研究[J].中国修复重建外科杂志,2017,31(3):351-356.
[13]王斌,王小琴.慢性间歇低氧对大鼠牙周炎模型血清及牙龈组织中白细胞介素-18和低氧诱导因子-1α的影响[J].华西口腔医学杂志,2015,33(4):383-387.