[1]熊辉,曲良烨,向黎黎,等.痛风性关节炎湿热证病证结合模型的建立[J].中医正骨,2014,26(03):14-20.
　Xiong Hui*,Qu Liangye,Xiang Lili,et al.A rat model of gouty arthritis combined with dampness-heat syndrome[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2014,26(03):14-20.
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痛风性关节炎湿热证病证结合模型的建立()
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《中医正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第26卷
期数:
2014年03期
页码:
14-20
栏目:
基础研究
出版日期:
2014-03-30

文章信息/Info

Title:
A rat model of gouty arthritis combined with dampness-heat syndrome
作者:
熊辉1; 曲良烨2; 向黎黎3; 齐新宇3; 陆小龙1; 郭玉星3
1.湖南中医药大学第二附属医院,湖南 长沙 410005;
2.张仲景国医院,河南 南阳 473000;
3.湖南中医药大学,湖南 长沙 410007
Author(s):
Xiong Hui*; Qu Liangye; Xiang Lili; Qi Xinyu; Lu Xiaolong; Guo Yuxing.
*The Second Affiliated Hospital of Hunan University of Traditional Chinese Medicine,Changsha 410005,Hunan,China
关键词:
关节炎; 痛风性 湿热 证候 模型; 动物
Keywords:
Arthritis; gouty;  Dampness-heat;  Symptom complex;  Models; animal
摘要:
目的:探讨建立痛风性关节炎湿热证病证结合大鼠模型的方法。方法:将40只成年健康雄性SD大鼠随机分为4组,每组10只,体质量(200±20)g。A、B组正常喂养,B组于造模开始后第13天在右侧踝关节腔注射微晶尿酸钠溶液。C、D组正常喂养,同时自由饮用200 g·L-1蜂蜜水,并分别以油脂(1 g·100g-1)或52%红星二锅头酒(1 mL·100 g-1)灌胃,二者交替进行,持续10 d。自第11天开始,置入温度(32±2)℃、相对湿度(92±3)%的人工气候箱中喂养。D组大鼠同时于第13天在右侧踝关节腔注射微晶尿酸钠溶液。造模期间观察大鼠的饮食、精神状态、大小便、舌象等情况; 造模开始后第15天,测定尿液水通道蛋白2含量; 造模开始后第16天,测定血浆内皮素和血浆降钙素基因相关肽含量; 造模结束后处死大鼠,观察舌组织和右侧踝关节滑膜组织的病理变化。结果:①一般情况观察结果。A组大鼠喂养期间,饮食、饮水、活动及大便均正常。B组大鼠造模前期饮食、饮水、活动及大便均正常,自第13天开始饮食、饮水减少,烦躁不安,右踝关节明显增粗,且红、肿、热,伴活动受限。C组大鼠开始造模后逐渐出现饮食、饮水量减少,嗜卧懒动、行动呆滞,毛发蓬松,颜色枯槁,小便黄; 造模第5天,开始出现大便溏泻或粘腻; 第11天出现明显呼吸粗重,烦躁不安,毛发疏松、粗糙,阴囊松弛下垂,大便溏泻或粘腻症状加重,渐见肛周污秽。D组大鼠造模后症状与C组相同,第13天开始合并出现B组造模后的相同表现。②尿液水通道蛋白2含量测定结果。4组大鼠尿液水通道蛋白2含量比较,差异有统计学意义[(0.251±0.018)mg·mL-1,(0.249±0.020)mg·mL-1,(0.233±0.014)mg·mL-1,(0.233±0.011)mg·mL-1,F=0.511,P=0.003]。A组与B组比较,差异无统计学意义(P=0.760); A组高于C组和D组(P=0.003; P=0.003); B组高于C组和D组(P=0.003; P=0.003); C组和D组比较,差异无统计学意义(P=0.751)。③血浆内皮素含量测定结果。4组大鼠血浆内皮素含量比较,差异有统计学意义[(4.14±0.08)pg·mL-1,(17.75±0.51)pg·mL-1,(19.01±0.13)pg·mL-1,(19.96±0.50)pg·mL-1,F=4.151,P=0.001]。A组低于B、C、D组(P=0.000; P=0.000; P=0.000); B组低于C组和D组(P=0.000; P=0.000); C组低于D组(P=0.000)。④血浆降钙素基因相关肽含量测定结果。4组大鼠血浆降钙素基因相关肽含量比较,差异有统计学意义[(91.29±0.42)pg·mL-1,(47.99±0.65)pg·mL-1,(56.32±2.17)pg·mL-1,(60.20±0.40)pg·mL-1,F=2.616,P=0.003]。A组高于B、C、D组(P=0.000; P=0.000; P=0.000); B组低于C组和D组(P=0.000; P=0.000); C组低于D组(P=0.000)。⑤血浆内皮素含量和降钙素基因相关肽含量的相关性分析结果。相关性分析结果显示,A、B组血浆内皮素含量和降钙素基因相关肽含量呈正相关(r=0.886,P=0.002; r=0.989,P=0.001); C、D组血浆内皮素含量和降钙素基因相关肽含量呈负相关(r=-0.706,P=0.005; r=-0.725,P=0.007)。⑥舌象及舌组织观察结果。A组大鼠舌淡红、苔薄白,B组舌红、苔微黄,C组舌红、苔腻微黄,D组舌红、苔黄腻。A组丝状乳头排列正常,无破坏,乳头呈圆锥形,尖端略向咽部倾斜。B组丝状乳头复层扁平上皮多有角化、脱落,且乳头高度参差不齐,尖端变钝或消失,上皮的厚度较C组变厚。菌状乳头固有层毛细血管增多,乳头数目及其分支减少。C组丝状乳头复层扁平上皮角化较B组严重,上皮形状矮短,尖端变钝或消失。菌状乳头固有层毛细血管增多,乳头数目及其分支增多。D组丝状乳头复层扁平上皮见角化、脱落,根部间或有空洞表现,上皮形状矮短并多有破坏,尖端变钝间或有消失; 菌状乳头固有层毛细血管增多。⑦踝关节滑膜组织观察结果。A组踝关节及其周围组织结构正常清晰,滑膜完整正常,无任何组织病理学改变; B组踝关节及其周围组织可见滑膜壁增厚,血管充血明显,滑膜及附近软骨被炎症细胞严重浸润破坏,出现组织坏死现象; C组表现同A组,D组表现同B组; 在局部解剖关节取材时,B、D组踝关节腔内可见尿酸盐结晶沉着。结论:在高脂高糖饮食、湿热环境、关节腔注射微晶尿酸钠溶液复合因素干预下,可复制出痛风性关节炎湿热证病证结合大鼠模型。
Abstract:
Objective:To explore the method of building a rat model of gouty arthritis combined with dampness-heat syndrome.Methods:Forty adult healthy male SD rats(weight 200+/-20 g)were randomly divided into 4 groups,10 rats in each group.Rats in group A and B were fed normally,and rats in group B were administered with intra-articular injection of microcrystalline sodium urate(MSU)solution in the right ankle on the 13th day after the beginning of modeling.Rats in group C and D were fed normally.Meanwhile,they could drink the honey water(200 g/L)freely and they were intragastric administrated with grease(1 g/100 g)and Chinese spirits(52%(vol/vol),1 mL/100 g)in turn for 10 consecutive days.Since the 11th day,they were fed in the artificial climate box(temperature,32+/-2 degrees centigrade; relative humidity,92+/-3%).Meanwhile,the rats in group D were administered with intra-articular injection of MSU solution in the right ankle on the 13th day.The rats's diet,mental status,urination and defecation and tongue demonstration were observed during modeling period.The urine contents of aquaporin-2(AQP2)were measured on the 15th day from the beginning of modeling.The plasma contents of endothelin(ET)and calcitonin generelated peptide(CGRP)were measured on the 16th day from the beginning of modeling.All rats were executed after the modeling and the pathological changes in tongue tissue and synovial tissue of right ankle joint were observed.Results:During feeding period,eating,drinking,activities and excrements of the rats in group A were normal.Eating,drinking,activities and excrements of the rats in group B were normal in the early period after the beginning of the modeling,while since the 13th day,their eating and drinking began to decrease and they became restless.Thickening,red,swelling,hot and action-limited were found in the right ankle joints.After the beginning of modeling,rats in group C presented with reduction in eating and drinking,indulging in lying,sluggish in movement,fluffy and dry hair and yellow urine.On the 5th day from the beginning of modeling,diarrhea or sticky excrements appeared for the rats.On the 11th day,the symptoms of rough breathing,restlessness,loose and rough in hair,flabby and sagging in scrotum and severe diarrhea appeared,and filth were found around the anus.The rats in group D presented with the same symptoms as those of group C after the beginning of modeling,and moreover,presented with the same symptoms as those of group B after the 13th day after modeling.There was statistical difference in the urine content of AQP2 between the 4 groups(0.251+/-0.018,0.249+/-0.020,0.233+/-0.014,0.233+/-0.011 mg/mL,F=0.511,P=0.003).There was no statistical difference between group A and group B(P=0.760).Group A surpassed group C and group D(P=0.003; P=0.003).Group B surpassed group C and group D(P=0.003; P=0.003).There was no statistical difference between group C and group D(P=0.751).There was statistical difference in the plasma content of ET between the 4 groups(4.14+/-0.08,17.75+/-0.51,19.01+/-0.13,19.96+/-0.50 pg/mL,F=4.151,P=0.001).The plasma content of ET was lower in group A compared with group B,C and D(P=0.000; P=0.000; P=0.000),and was lower in group B compared with group C and group D(P=0.000; P=0.000),and was lower in group C compared with group D(P=0.000).There was statistical difference in the plasma content of CGRP between the 4 groups(91.29+/-0.42,47.99+/-0.65,56.32+/-2.17,60.20+/-0.40 pg/mL,F=2.616,P=0.003).The plasma content of CGRP was higher in group A compared with group B,C and D(P=0.000; P=0.000; P=0.000),and was lower in group B compared with group C and group D(P=0.000; P=0.000),and was lower in group C compared with group D(P=0.000).The results of correlation analysis on the plasma content of ET and CGRP showed that they were positively correlated with each other in group A and group B(r=0.886,P=0.002; r=0.989,P=0.001)and were negatively correlated with each other in group C and group D(r=-0.706,P=0.005; r=-0.725,P=0.007).The results of observation on tongue demonstration and tongue tissue showed that the rats in group A presented with pink tongue and thin white coated tongue,the rats in group B presented with red tongue and yellowish coated tongue,the rats in group C presented with red tongue and yellowish greasy coated tongue,and the rats in group D presented with red tongue and yellow greasy coated tongue.The filiform papillae of the tongue in group A arranged regularly and was not damaged,and the papillas were conical and its tip leaned to the pharynx slightly.The filiform papillae of the tongue in group B presented with stratified squamous epithelium cornification and fallen off,the heights of the papillas were irregular,the tips became blunt or disappeared,and the thickness of epithele became thickening compared with that of group C.The capillaries increased in the lamina propria of the fungiform papillae,and the papillae and its branches decreased.Compared to group B,the keratinization of stratified squamous epithelium of filiform papillae were severer in group C,the epithel was short and its tips were blunt or even disappeared.The capillaries increased in the lamina propria of the fungiform papillae,and the fungiform papillae and its branches increased.The stratified squamous epithelium of filiform papillae of group D were keratinized and fallen off,with cavity in the roots.The epithel was short and was damaged and its tips were blunt or even disappeared.The capillaries increased in the lamina propria of the fungiform papillae.The results of observation on synovial tissue of the ankle joint showed that the ankle joints and their surrounding tissues structure were normal and clear in rats of group A,and the synovium were complete and normal without any histopathological changes.The thickened synovial membrane walls and obvious vascular engorgement were found in ankle joints and their surrounding tissues in group B,and the synovial membrane and their vicinal cartilages were infiltrated and damaged by inflammatory cells,and meanwhile,the tissue necrosis were found.The manifestation of group C was the same as that of group A,and the manifestation of group D was the same as that of group B.The urate crystals were found in ankle joint cavities in group B and group D.Conclusion:The rat model of gouty arthritis combined with dampness-heat syndrome could be built through intervention of composite factors including high fat and high glucose diet,wet heat environment and intra-articular injection of MSU solution.

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备注/Memo

备注/Memo:
基金项目:湖南省中医药科研计划项目(2010025),湖南省自然科学基金项目(13JJ3102)
通讯作者:曲良烨 E-mail:quliangye2013@sina.com
更新日期/Last Update: 2014-03-20