云南白药对实验性牙周炎及实验性糖尿病牙周炎抗炎作用的研究
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摘要
【目的】通过动物实验客观评价云南白药对实验性牙周炎及实验性糖尿病牙周炎的抗炎作用,进一步探讨云南白药对炎症反应的作用机制。
【方法】选用6周龄健康雄性SD大鼠75只,将实验动物随机分为三组:空白对照组(N组=15只)、糖尿病牙周炎组(DP组=30只)、单纯牙周炎组(P组=30只)。DP组采用一次性注射STZ的方法诱导大鼠糖尿病模型,然后DP组和P组采用丝线结扎的方法建立牙周炎模型。模型建立后,P组大鼠随机分为牙周炎实验组(P1组,n=15)和牙周炎对照组(P2组,n=15);DP组随机分为糖尿病牙周炎实验组(DP1组,n=15),糖尿病牙周炎对照组(DP2组,n=15)。分别对P1组和DP1组大鼠每日云南白药1ml(50mg/ml)灌胃,P2组和DP2组每日蒸馏水1ml灌胃。分别在2wk、4wk与6wk处死。处死前通过心脏采血用于TNF-α、IL-1和AGEs的检测。处死后取大鼠上颌骨,4%多聚甲醛固定,用于TNF-α的免疫组化检测。观测指标包括:(1)临床指标:牙周探诊深度(PD),牙槽骨丧失(ABL);(2)组织学指标:牙周组织中TNF-α的光密度值测定分析;(3)采用ELISA方法检测外周血血清中IL-1β和TNF-α水平的变化。(4)血清中AGEs含量测定。资料采用单因素方差分析统计学处理。
【结果】实验性牙周炎及实验性糖尿病牙周炎SD大鼠灌胃云南白药(50mg/ml)能减轻牙周炎症。在4wk、6wk时,大鼠血清中TNF-α、IL-1β的含量、牙槽骨中TNF-α的含量、牙周探诊深度均为P1组＜P2组、DP1组＜DP2组;在6wk时,牙槽骨丧失P1组＜P2组、DP1组＜DP2组以及血清中AGEs含量DP1组＜DP2组。
【结论】云南白药对动物实验性牙周炎及实验性糖尿病牙周炎的炎症反应和牙周骨组织的破坏有一定的抑制作用.并且在一定程度上对牙槽骨中的细胞因子TNF-α的表达亦有抑制作用,同时还能降低血清中TNF-α、IL-1β的水平,可能对DP组中血清中AGEs有抑制作用。
Objective:To evaluate the anti-inflammation effects of Yunnan Baiyao on experimental periodontitis and experimental diabetes periodontitis,and explore it's possible in vitro mechanism of the inflammatory response.
Methods:Seventy-five 6-week-old healthy male SD rats were involed in this study, the experimental animals were randomly divided into three groups:control group(N group=15rats),diabetic periodontitis group(DP group=30rats),periodontitis without diabetes group(P group = 30rats).DP group was injected STZ to induce diabetes model,after then both DP group and P group were silk ligatied to establish the periodontitis model.When model was established,periodontitis rats were randomly divided into experimental group(P1 group,n=15rats) and the periodontitis control group(P2 group,n=15rats);DP group rats were randomly divided into experimental diabetic periodontitis(DP1 group,n=15rats),and diabetic periodontitis control group (DP2 group,n=15rats).Rats in P1 group and DP1 group,Yunnan Baiyao 1ml (50mg/ml) were gavaged daily respectively rats,and the rats were gavaged 1ml of distilled water per day in P2 group and DP2 group.Taking the blood sample from the heart for test TNF-α,IL-1 and AGEs.After the rats dead,taking the maxilla bone sample with 4%paraformaldehyde,and the sample will be used to test TNF-αby immunohistochemistry.Observed indicators include:(1) clinical parameters:probing depth(PD),alveolar bone loss(ABL);(2) histological indicators:measure and analysis TNF-αoptical density in periodontal tissue;(3) Serum IL-1βand TNF-αlevel changes were analysis by ELISA.(4) The determination of serum AGEs.Data using single-factor analysis of variance statistically.
Results:Experimental periodontitis and experimental diabetes periodontitis SD rats gastric lavage periodontitis(including Yunnan Baiyao 50mg/ml) can reduce the periodontal inflammation.In 4wk and 6wk,the content of TNF-αand IL-1βin rats serum,the content of TNF-αalveolar bone,periodontal probing depth of appointment all express:P1group＜P2 group,DP1 group＜DP2 group.In 6wk,alveolar bone loss P1 group＜DP1 group,P2 group＜DP2group,and the content of AGEs in serum DP1 group＜DP2 group.
Conclusions:Yunnan Baiyao can inhibite inflammation response of animal experimental periodontitis and experimental diabetes periodontitis,and also can reduce the destruction of periodontal tissue.the Chinese medicine also have same function to inhibite TNF-αexpression in alveolar bone,at the same time,it reduce the content of serum TNF-αand IL-1β.We also know it's possibility inhibition function for the serum AGEs.
【方法】选用6周龄健康雄性SD大鼠75只,将实验动物随机分为三组:空白对照组(N组=15只)、糖尿病牙周炎组(DP组=30只)、单纯牙周炎组(P组=30只)。DP组采用一次性注射STZ的方法诱导大鼠糖尿病模型,然后DP组和P组采用丝线结扎的方法建立牙周炎模型。模型建立后,P组大鼠随机分为牙周炎实验组(P1组,n=15)和牙周炎对照组(P2组,n=15);DP组随机分为糖尿病牙周炎实验组(DP1组,n=15),糖尿病牙周炎对照组(DP2组,n=15)。分别对P1组和DP1组大鼠每日云南白药1ml(50mg/ml)灌胃,P2组和DP2组每日蒸馏水1ml灌胃。分别在2wk、4wk与6wk处死。处死前通过心脏采血用于TNF-α、IL-1和AGEs的检测。处死后取大鼠上颌骨,4%多聚甲醛固定,用于TNF-α的免疫组化检测。观测指标包括:(1)临床指标:牙周探诊深度(PD),牙槽骨丧失(ABL);(2)组织学指标:牙周组织中TNF-α的光密度值测定分析;(3)采用ELISA方法检测外周血血清中IL-1β和TNF-α水平的变化。(4)血清中AGEs含量测定。资料采用单因素方差分析统计学处理。
【结果】实验性牙周炎及实验性糖尿病牙周炎SD大鼠灌胃云南白药(50mg/ml)能减轻牙周炎症。在4wk、6wk时,大鼠血清中TNF-α、IL-1β的含量、牙槽骨中TNF-α的含量、牙周探诊深度均为P1组＜P2组、DP1组＜DP2组;在6wk时,牙槽骨丧失P1组＜P2组、DP1组＜DP2组以及血清中AGEs含量DP1组＜DP2组。
【结论】云南白药对动物实验性牙周炎及实验性糖尿病牙周炎的炎症反应和牙周骨组织的破坏有一定的抑制作用.并且在一定程度上对牙槽骨中的细胞因子TNF-α的表达亦有抑制作用,同时还能降低血清中TNF-α、IL-1β的水平,可能对DP组中血清中AGEs有抑制作用。
Objective:To evaluate the anti-inflammation effects of Yunnan Baiyao on experimental periodontitis and experimental diabetes periodontitis,and explore it's possible in vitro mechanism of the inflammatory response.
Methods:Seventy-five 6-week-old healthy male SD rats were involed in this study, the experimental animals were randomly divided into three groups:control group(N group=15rats),diabetic periodontitis group(DP group=30rats),periodontitis without diabetes group(P group = 30rats).DP group was injected STZ to induce diabetes model,after then both DP group and P group were silk ligatied to establish the periodontitis model.When model was established,periodontitis rats were randomly divided into experimental group(P1 group,n=15rats) and the periodontitis control group(P2 group,n=15rats);DP group rats were randomly divided into experimental diabetic periodontitis(DP1 group,n=15rats),and diabetic periodontitis control group (DP2 group,n=15rats).Rats in P1 group and DP1 group,Yunnan Baiyao 1ml (50mg/ml) were gavaged daily respectively rats,and the rats were gavaged 1ml of distilled water per day in P2 group and DP2 group.Taking the blood sample from the heart for test TNF-α,IL-1 and AGEs.After the rats dead,taking the maxilla bone sample with 4%paraformaldehyde,and the sample will be used to test TNF-αby immunohistochemistry.Observed indicators include:(1) clinical parameters:probing depth(PD),alveolar bone loss(ABL);(2) histological indicators:measure and analysis TNF-αoptical density in periodontal tissue;(3) Serum IL-1βand TNF-αlevel changes were analysis by ELISA.(4) The determination of serum AGEs.Data using single-factor analysis of variance statistically.
Results:Experimental periodontitis and experimental diabetes periodontitis SD rats gastric lavage periodontitis(including Yunnan Baiyao 50mg/ml) can reduce the periodontal inflammation.In 4wk and 6wk,the content of TNF-αand IL-1βin rats serum,the content of TNF-αalveolar bone,periodontal probing depth of appointment all express:P1group＜P2 group,DP1 group＜DP2 group.In 6wk,alveolar bone loss P1 group＜DP1 group,P2 group＜DP2group,and the content of AGEs in serum DP1 group＜DP2 group.
Conclusions:Yunnan Baiyao can inhibite inflammation response of animal experimental periodontitis and experimental diabetes periodontitis,and also can reduce the destruction of periodontal tissue.the Chinese medicine also have same function to inhibite TNF-αexpression in alveolar bone,at the same time,it reduce the content of serum TNF-αand IL-1β.We also know it's possibility inhibition function for the serum AGEs.
引文
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[1]丁志锋,段金成,和红兵.根管治疗术后根尖微渗漏影响因素的研究现状.中国实用口腔科杂志,2009,2(11):695-7.
[2]段金成,丁志锋,和红兵.炎症反应介质对炎症影响机制的探讨.昆明医学院学报,2009,30(3B):261-3.
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