独一味胶囊对复发性口腔溃疡大鼠血清TNF-α、IL-2和IL-6水平的影响
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摘要
目的探讨独一味胶囊对复发性口腔溃疡(ROU)大鼠血清肿瘤坏死因子(TNF-α)、白介素2(IL-2)及白介素6(IL-6)水平的影响。方法 SPF级雄性SD大鼠40只,采用注射完全弗氏佐剂法建立ROU大鼠模型,随机分为4组(低、中、高剂量组及对照组),每组10只。低、中、高剂量组大鼠以1mL/100g灌服低、中、高剂量独一味胶囊溶液(独一味胶囊内容物以无菌生理盐水配置浓度为5%、10%、15%的溶液),对照组灌服1mL/100g无菌生理盐水。各组分别在给药1、3、7d后,对比各组大鼠口腔溃疡数目、溃疡面积,然后尾静脉采血,采用酶联免疫吸附实验(ELISA)检测各组血清TNF-α、IL-2及IL-6水平,并对比各组数据;给药7d后处死大鼠取溃疡组织,采用免疫印迹试验检测各组口腔溃疡组织中TNF-α、IL-2及IL-6蛋白相对表达量,并对比各组数据。结果低、中、高剂量组大鼠给药后不同时刻溃疡数目、溃疡面积、血清TNF-α、IL-2及IL-6水平组间、时间及交互比较,低、中、高剂量组大鼠给药7d后溃疡黏膜组织TNF-α、IL-2及IL-6蛋白相对表达量比较,差异有统计学意义(P<0.05)。低、中、高剂量组给药后不同时刻溃疡数目、溃疡面积、血清TNF-α、IL-6水平及给药7d后溃疡黏膜组织中TNF-α、IL-6蛋白相对表达量均显著低于对照组(P<0.05),且低、中、高剂量组均随时间及剂量依赖性降低(P<0.05);低、中、高剂量组给药后不同时刻血清IL-2水平及给药7d后溃疡黏膜组织中IL-2蛋白相对表达量均显著高于对照组(P<0.05),且3实验组均随时间及剂量依赖性升高(P<0.05);对照组大鼠给药后不同时刻溃疡数目、溃疡面积、TNF-α、IL-2、IL-6血清水平及给药7d后口腔溃疡黏膜组织中TNF-α、IL-2、IL-6蛋白相对表达量差异无统计学意义(P>0.05)。结论独一味胶囊可显著减少大鼠口腔溃疡数目及溃疡面积,加速大鼠口腔溃疡愈合,推测与独一味胶囊显著下调TNF-α、IL-6在血清及口腔溃疡黏膜组织中的表达及上调IL-2的表达有关。
Objective To explore the effects of the Duyiwei capsules on tumor necrosis factor-α(TNF-α),interleukin-2(IL-2)and interleukin-6(IL-6)levels of serum in rats with recurrent oral ulcer(ROU).Methods There were 40 males SD rats with SPF,and the rats model of ROU were established by injection of complete Freund's adjuvant method,which were randomly divided into 4 groups,10 in each group.The rats of the low,medium and high dose group were given 1 mL/100 g of different concentrations of the Duyiwei capsules solution,which were prepared by using sterile saline solution with concentration of 5%,10% and 15% of the Duyiwei capsules contents.The numbers and areas of oral ulcer of rats were observed and compared after 1,3 and 7 days.The TNF-α,IL-2 and IL-6 levels of serum at 1,3 and 7 days were detected by enzyme-linked immunosorbent assay(ELISA).The rats were executed after 7 days and the ulcer tissues were taken.The relative expression of TNF-α,IL-2 and IL-6 proteins of the tissues were detected by Western Blotting.Results There were significant differences in the numbers and areas of oral ulcer,the serum levels of TNF-α,IL-2 and IL-6 among the 3 doses groups,moments and interactions(P <0.05).The protein levels of TNF-α,IL-2 and IL-6 in the mucosal tissues after 7 days of the 3 doses groups were significantly different(P <0.05).The numbers and areas of oral ulcer,the serum levels of TNF-αand IL-6 at 1,3 and 7 days and the protein levels of TNF-αand IL-6 in the mucosal tissues after 7 days of the 3 doses groups were significantly lower than those of the control group(P <0.05),every dose group showed a significant decrease with time and dose-dependence(P <0.05).The serum levels of IL-2 at 1,3 and 7 days and the protein levels of IL-2 in the mucosal tissues after 7 days of the 3 doses groups were significantly higher than those of the control group(P <0.05),every dose group showed a significant increase with time and dose-dependence(P <0.05).There were no significant differences between the numbers and areas of oral ulcer,the serum levels of TNF-α,IL-2 and IL-6 after drug delivery of 1,3 and 7 days and the protein levels of TNF-α,IL-2 and IL-6 in the mucosal tissues after 7 days of the 3 dose groups(P >0.05).Conclusion The Duyiwei capsules can significantly reduce the numbers and areas of oral ulcer of rats,also accelerate the healing of oral ulcer in rats,and it may be related to the levels of the serum and protein of TNF-αand IL-6 down-regulation and the serum and protein of IL-2 up-regulation by the Duyiwei capsules.
Objective To explore the effects of the Duyiwei capsules on tumor necrosis factor-α(TNF-α),interleukin-2(IL-2)and interleukin-6(IL-6)levels of serum in rats with recurrent oral ulcer(ROU).Methods There were 40 males SD rats with SPF,and the rats model of ROU were established by injection of complete Freund's adjuvant method,which were randomly divided into 4 groups,10 in each group.The rats of the low,medium and high dose group were given 1 mL/100 g of different concentrations of the Duyiwei capsules solution,which were prepared by using sterile saline solution with concentration of 5%,10% and 15% of the Duyiwei capsules contents.The numbers and areas of oral ulcer of rats were observed and compared after 1,3 and 7 days.The TNF-α,IL-2 and IL-6 levels of serum at 1,3 and 7 days were detected by enzyme-linked immunosorbent assay(ELISA).The rats were executed after 7 days and the ulcer tissues were taken.The relative expression of TNF-α,IL-2 and IL-6 proteins of the tissues were detected by Western Blotting.Results There were significant differences in the numbers and areas of oral ulcer,the serum levels of TNF-α,IL-2 and IL-6 among the 3 doses groups,moments and interactions(P <0.05).The protein levels of TNF-α,IL-2 and IL-6 in the mucosal tissues after 7 days of the 3 doses groups were significantly different(P <0.05).The numbers and areas of oral ulcer,the serum levels of TNF-αand IL-6 at 1,3 and 7 days and the protein levels of TNF-αand IL-6 in the mucosal tissues after 7 days of the 3 doses groups were significantly lower than those of the control group(P <0.05),every dose group showed a significant decrease with time and dose-dependence(P <0.05).The serum levels of IL-2 at 1,3 and 7 days and the protein levels of IL-2 in the mucosal tissues after 7 days of the 3 doses groups were significantly higher than those of the control group(P <0.05),every dose group showed a significant increase with time and dose-dependence(P <0.05).There were no significant differences between the numbers and areas of oral ulcer,the serum levels of TNF-α,IL-2 and IL-6 after drug delivery of 1,3 and 7 days and the protein levels of TNF-α,IL-2 and IL-6 in the mucosal tissues after 7 days of the 3 dose groups(P >0.05).Conclusion The Duyiwei capsules can significantly reduce the numbers and areas of oral ulcer of rats,also accelerate the healing of oral ulcer in rats,and it may be related to the levels of the serum and protein of TNF-αand IL-6 down-regulation and the serum and protein of IL-2 up-regulation by the Duyiwei capsules.
引文
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[12]李增清,杨东新,周真,等.白介素6受体基因rs4129267位点多态性与汉族儿童哮喘易患性及血清IgE水平的关联研究[J].中国全科医学,2016,19(36):4474-4477.
[13]OKAMOTO M,MIZUKAMI Y.GPER negatively regulates TNFα-induced IL-6production in human breast cancer cells via NF-κB pathway[J].Endocr J,2016,63(5):485-493.
[14]乙永林,孙连先.HPLC法同时测定藏药独一味中山栀苷甲酯、木犀草素、芹菜素的含量[J].中国药房,2016,27(33):4733-4735.
[15]黄逸婷,张梅.白介素-2和白介素-2受体的调控在自身免疫性疾病和器官移植中的作用[J].免疫学杂志,2017,15(10):69-71.
[16]常莉,胡炯.IL-2调控免疫耐受及其在慢性移植物抗宿主病治疗中的应用研究[J].世界临床药物,2016,37(9):588-592.
[17]贾诩,李茂星,陶锐,等.藏药独一味中的主要化学成分环烯醚萜苷化合物在动物的镇痛抗炎作用研究[J].中国临床药理学杂志,2015,31(8):635-639.
[2]GUPAT S,JAWANDA M K.Oral lichen planus:an update on etiology,pathogenesis,clinical presentation,diagnosis and management[J].Indian J Dermatol,2015,60(3):222-225.
[3]白蓉,郭延伟.独一味中苯乙醇苷提取物的镇痛抗炎作用及毒性研究[J].华西药学杂志,2015,30(3):384-385.
[4]梁晓燕,高婷,蒋建芳.独一味胶囊用于下肢骨折患者镇痛止血的效果观察[J].西北国防医学杂志,2016,54(11):753-754.
[5]张维国,王助英.医用臭氧关节腔注射联合独一味滴丸治疗膝骨关节炎的疗效分析[J].药物评价研究,2017,40(1):112-114.
[6]LALABONVA H.Clinical assessment of the therapeutic effect of low level laser therapy on chronic recurrent aphthous stomatitis[J].Biotechnol Biotec Eq,2014,28(5):929-933.
[7]张一凡,覃璇,林娟,等.左旋咪唑或联合中药治疗复发性阿弗他口腔溃疡的系统评价与Meta分析[J].口腔医学研究,2015,44(5):479-484.
[8]徐斌,石宇远,汪柳静,等.中药独一味口腔贴膜的研制及其在口腔溃疡治疗中的临床应用效果探讨[J].中华中医药学刊,2016,19(7):1666-1668.
[9]韩燕,刘沛源,肖京.千金口疮加味颗粒治疗阴虚火旺型复发性口腔溃疡的临床观察[J].中国中西医结合杂志,2015,35(7):816-819.
[10]刘调玲.独一味胶囊治疗慢性牙髓炎75例[J].西部中医药,2016,29(8):90-91.
[11]HAJEER A H,Hutchinson I V.TNF‐αgene polymorphism:Clinical and biological implications.[J].Microsc Res Techn,2015,50(3):216-228.
[12]李增清,杨东新,周真,等.白介素6受体基因rs4129267位点多态性与汉族儿童哮喘易患性及血清IgE水平的关联研究[J].中国全科医学,2016,19(36):4474-4477.
[13]OKAMOTO M,MIZUKAMI Y.GPER negatively regulates TNFα-induced IL-6production in human breast cancer cells via NF-κB pathway[J].Endocr J,2016,63(5):485-493.
[14]乙永林,孙连先.HPLC法同时测定藏药独一味中山栀苷甲酯、木犀草素、芹菜素的含量[J].中国药房,2016,27(33):4733-4735.
[15]黄逸婷,张梅.白介素-2和白介素-2受体的调控在自身免疫性疾病和器官移植中的作用[J].免疫学杂志,2017,15(10):69-71.
[16]常莉,胡炯.IL-2调控免疫耐受及其在慢性移植物抗宿主病治疗中的应用研究[J].世界临床药物,2016,37(9):588-592.
[17]贾诩,李茂星,陶锐,等.藏药独一味中的主要化学成分环烯醚萜苷化合物在动物的镇痛抗炎作用研究[J].中国临床药理学杂志,2015,31(8):635-639.