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[1]熊輝,曲良燁,向黎黎,等.痛風(fēng)性關(guān)節(jié)炎濕熱證病證結(jié)合模型的建立[J].中醫(yī)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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痛風(fēng)性關(guān)節(jié)炎濕熱證病證結(jié)合模型的建立()
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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第26卷
期數(shù):
2014年03期
頁(yè)碼:
14-20
欄目:
基礎(chǔ)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:
熊輝1; 曲良燁2; 向黎黎3; 齊新宇3; 陸小龍1; 郭玉星3
1.湖南中醫(yī)藥大學(xué)第二附屬醫(yī)院,湖南 長(zhǎng)沙 410005;
2.張仲景國(guó)醫(yī)院,河南 南陽(yáng) 473000;
3.湖南中醫(yī)藥大學(xué),湖南 長(zhǎng)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
關(guān)鍵詞:
關(guān)節(jié)炎; 痛風(fēng)性 濕熱 證候 模型; 動(dòng)物
Keywords:
Arthritis; gouty;  Dampness-heat;  Symptom complex;  Models; animal
摘要:
目的:探討建立痛風(fēng)性關(guān)節(jié)炎濕熱證病證結(jié)合大鼠模型的方法。方法:將40只成年健康雄性SD大鼠隨機(jī)分為4組,每組10只,體質(zhì)量(200±20)g。A、B組正常喂養(yǎng),B組于造模開(kāi)始后第13天在右側(cè)踝關(guān)節(jié)腔注射微晶尿酸鈉溶液。C、D組正常喂養(yǎng),同時(shí)自由飲用200 g·L-1蜂蜜水,并分別以油脂(1 g·100g-1)或52%紅星二鍋頭酒(1 mL·100 g-1)灌胃,二者交替進(jìn)行,持續(xù)10 d。自第11天開(kāi)始,置入溫度(32±2)℃、相對(duì)濕度(92±3)%的人工氣候箱中喂養(yǎng)。D組大鼠同時(shí)于第13天在右側(cè)踝關(guān)節(jié)腔注射微晶尿酸鈉溶液。造模期間觀察大鼠的飲食、精神狀態(tài)、大小便、舌象等情況; 造模開(kāi)始后第15天,測(cè)定尿液水通道蛋白2含量; 造模開(kāi)始后第16天,測(cè)定血漿內(nèi)皮素和血漿降鈣素基因相關(guān)肽含量; 造模結(jié)束后處死大鼠,觀察舌組織和右側(cè)踝關(guān)節(jié)滑膜組織的病理變化。結(jié)果:①一般情況觀察結(jié)果。A組大鼠喂養(yǎng)期間,飲食、飲水、活動(dòng)及大便均正常。B組大鼠造模前期飲食、飲水、活動(dòng)及大便均正常,自第13天開(kāi)始飲食、飲水減少,煩躁不安,右踝關(guān)節(jié)明顯增粗,且紅、腫、熱,伴活動(dòng)受限。C組大鼠開(kāi)始造模后逐漸出現(xiàn)飲食、飲水量減少,嗜臥懶動(dòng)、行動(dòng)呆滯,毛發(fā)蓬松,顏色枯槁,小便黃; 造模第5天,開(kāi)始出現(xiàn)大便溏瀉或粘膩; 第11天出現(xiàn)明顯呼吸粗重,煩躁不安,毛發(fā)疏松、粗糙,陰囊松弛下垂,大便溏瀉或粘膩癥狀加重,漸見(jiàn)肛周污穢。D組大鼠造模后癥狀與C組相同,第13天開(kāi)始合并出現(xiàn)B組造模后的相同表現(xiàn)。②尿液水通道蛋白2含量測(cè)定結(jié)果。4組大鼠尿液水通道蛋白2含量比較,差異有統(tǒng)計(jì)學(xué)意義[(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組與B組比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P=0.760); A組高于C組和D組(P=0.003; P=0.003); B組高于C組和D組(P=0.003; P=0.003); C組和D組比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P=0.751)。③血漿內(nèi)皮素含量測(cè)定結(jié)果。4組大鼠血漿內(nèi)皮素含量比較,差異有統(tǒng)計(jì)學(xué)意義[(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組低于B、C、D組(P=0.000; P=0.000; P=0.000); B組低于C組和D組(P=0.000; P=0.000); C組低于D組(P=0.000)。④血漿降鈣素基因相關(guān)肽含量測(cè)定結(jié)果。4組大鼠血漿降鈣素基因相關(guān)肽含量比較,差異有統(tǒng)計(jì)學(xué)意義[(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組高于B、C、D組(P=0.000; P=0.000; P=0.000); B組低于C組和D組(P=0.000; P=0.000); C組低于D組(P=0.000)。⑤血漿內(nèi)皮素含量和降鈣素基因相關(guān)肽含量的相關(guān)性分析結(jié)果。相關(guān)性分析結(jié)果顯示,A、B組血漿內(nèi)皮素含量和降鈣素基因相關(guān)肽含量呈正相關(guān)(r=0.886,P=0.002; r=0.989,P=0.001); C、D組血漿內(nèi)皮素含量和降鈣素基因相關(guān)肽含量呈負(fù)相關(guān)(r=-0.706,P=0.005; r=-0.725,P=0.007)。⑥舌象及舌組織觀察結(jié)果。A組大鼠舌淡紅、苔薄白,B組舌紅、苔微黃,C組舌紅、苔膩微黃,D組舌紅、苔黃膩。A組絲狀乳頭排列正常,無(wú)破壞,乳頭呈圓錐形,尖端略向咽部?jī)A斜。B組絲狀乳頭復(fù)層扁平上皮多有角化、脫落,且乳頭高度參差不齊,尖端變鈍或消失,上皮的厚度較C組變厚。菌狀乳頭固有層毛細(xì)血管增多,乳頭數(shù)目及其分支減少。C組絲狀乳頭復(fù)層扁平上皮角化較B組嚴(yán)重,上皮形狀矮短,尖端變鈍或消失。菌狀乳頭固有層毛細(xì)血管增多,乳頭數(shù)目及其分支增多。D組絲狀乳頭復(fù)層扁平上皮見(jiàn)角化、脫落,根部間或有空洞表現(xiàn),上皮形狀矮短并多有破壞,尖端變鈍間或有消失; 菌狀乳頭固有層毛細(xì)血管增多。⑦踝關(guān)節(jié)滑膜組織觀察結(jié)果。A組踝關(guān)節(jié)及其周圍組織結(jié)構(gòu)正常清晰,滑膜完整正常,無(wú)任何組織病理學(xué)改變; B組踝關(guān)節(jié)及其周圍組織可見(jiàn)滑膜壁增厚,血管充血明顯,滑膜及附近軟骨被炎癥細(xì)胞嚴(yán)重浸潤(rùn)破壞,出現(xiàn)組織壞死現(xiàn)象; C組表現(xiàn)同A組,D組表現(xiàn)同B組; 在局部解剖關(guān)節(jié)取材時(shí),B、D組踝關(guān)節(jié)腔內(nèi)可見(jiàn)尿酸鹽結(jié)晶沉著。結(jié)論:在高脂高糖飲食、濕熱環(huán)境、關(guān)節(jié)腔注射微晶尿酸鈉溶液復(fù)合因素干預(yù)下,可復(fù)制出痛風(fēng)性關(guān)節(jié)炎濕熱證病證結(jié)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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備注/Memo

備注/Memo:
基金項(xiàng)目:湖南省中醫(yī)藥科研計(jì)劃項(xiàng)目(2010025),湖南省自然科學(xué)基金項(xiàng)目(13JJ3102)
通訊作者:曲良燁 E-mail:[email protected]
更新日期/Last Update: 2014-03-20