本文選題：高尿酸血癥 + 虎杖苷��； 參考：《廣東藥科大學(xué)》2017年碩士論文

【摘要】：高尿酸血癥是困擾人類的眾多代謝疾病之一。大量的國內(nèi)外研究顯示,高尿酸血與許多疾病密切相關(guān),如急性痛風(fēng)性關(guān)節(jié)炎、尿酸性腎病、高血壓、肥胖、糖尿病和心血管疾病等。伴隨著我國經(jīng)濟(jì)水平的高速列車,人民的生活習(xí)慣,飲食規(guī)律發(fā)生了翻天覆地的變化。夜宵、快餐文化、啤酒、飲料(含果糖)等在我國的大肆泛濫,致使我國的高尿酸血癥患者爆發(fā)式的增長,并呈現(xiàn)低齡化的趨勢。嚴(yán)重影響著我國人民的身體健康以及生活質(zhì)量。目前高尿酸血癥的研究尚缺乏公認(rèn)的合理的動物模型和安全有效的治療藥物,故本次實驗我們希望嘗試建立一種血尿酸值較為平緩、穩(wěn)定、持久、對腎臟損害小的大鼠高尿酸血癥模型,并在此基礎(chǔ)上探究虎桂藥對的組分——虎杖苷和桂皮醛相互配伍對高尿酸血癥的治療作用以及其作用機(jī)制。1.大鼠高尿酸血癥模型實驗?zāi)康?優(yōu)化次黃嘌呤,氧嗪酸鉀,乙胺丁醇三種藥物聯(lián)合制備大鼠高尿酸血癥模型的使用劑量,以期得到更合適的大鼠高尿酸血癥模型。方法:雄性SD大鼠32只,隨機(jī)分為4組,每組8只。模型Ⅰ組:給以灌胃75mg/kg次黃嘌呤和250mg/kg鹽酸乙胺丁醇,皮下注射200mg/kg氧嗪酸鉀(1次/d)。模型Ⅱ組:給以灌胃75mg/kg次黃嘌呤和250mg/kg鹽酸乙胺丁醇(1次/d),皮下注射100mg/kg氧嗪酸鉀(1次/12h)。模型Ⅲ組:給以灌胃50mg/kg次黃嘌呤和250mg/kg鹽酸乙胺丁醇,皮下注射200mg/kg氧嗪酸鉀(1次/d)。空白組給予生理鹽水灌胃及橄欖油皮下注射。連續(xù)造模24天。第4,11,18d皮下注射后6h,大鼠眼眶取血,測SUA含量。第24天皮下注射后,分別3、6、12、24h取血測SUA含量,并測6h血清中BUN、Cr的含量。大鼠處死取腎,測察其病理改變。結(jié)果:與空白組對比,三組造模方式均能引起大鼠的血尿酸含量明顯上升(P0.01),可維持12h,尤其是模型Ⅱ組能達(dá)到24h;三組造模方式的大鼠腎臟病理切片表明,三種造模方式均能引起腎臟輕微損傷;模型Ⅰ組和模型Ⅱ組的血尿素氮含量與空白組對比明顯的升高(P0.01,P0.05)。結(jié)論:三種造模方式均能成功制備大鼠高尿酸血癥模型,并伴有輕微的腎臟器質(zhì)和功能損傷(尤其是模型Ⅲ組損傷最輕),具有血尿酸值增長維持時間長的優(yōu)點(diǎn)(尤其是模型Ⅱ組可持續(xù)24h)。2.虎桂藥對組分配伍對高尿酸血癥的治療及機(jī)制的探究目的:探究虎桂藥對的組分——虎杖苷(Polydatin)和桂皮醛(Cinnamaldehyde)互相配伍后,對大鼠高尿酸血癥的降尿酸作用,并從尿酸的排泄和生成兩個途徑,初步探索其多靶點(diǎn)降尿酸的機(jī)理,為中醫(yī)藥治療高尿酸血癥提供新的思路和方法。方法:雄性SD大鼠70只隨機(jī)分為7組�？瞻捉M、模型組、別嘌呤醇組、苯溴馬隆組、虎杖苷和桂皮醛(PC)的低、中、高組,每組10只。皮下注射氧嗪酸鉀(200mg/kg),7h后灌胃乙胺丁醇(250mg/kg)和次黃嘌呤(50mg/kg)制備高尿酸血癥大鼠模型,連續(xù)26天(1次/d)。第4天,皮下注射后6h眼眶取血測大鼠的SUA值。第5天起,皮下注射前1h,灌胃治療藥,持續(xù)21天(1次/d)。第25天將大鼠放進(jìn)代謝籠,收集尿液24h,備用。最后一天皮下注射6h后,大鼠腹主動脈取血,測定SUA、Cr、BUN和GPT的含量;取出大鼠肝,腎,小腸備用。ELISA試劑盒測定尿液中Kim-1、NGAL的含量;腎臟組織的病理學(xué)觀察,統(tǒng)計評分;qPCR測定肝臟XOD、ADA和腎臟GLUT9、URAT1、OAT1、OCT1的基因表達(dá);Western Blotting測定肝臟XOD和腎臟GLUT9、URAT1的蛋白表達(dá);免疫組化測定小腸MRP4的蛋白表達(dá),分析照片陽性的累積光密度值(IOD)。結(jié)果:造模后,大鼠SUA含量明顯升高(P0.01)。給予治療藥后,PC中、高組SUA含量明顯下降(P0.01,P0.05);PC中組BUN含量明顯下降(P0.05);PC三組的NGAL含量明顯下降(P0.01,P0.05);PC中、高組的Kim-1含量明顯下降(P0.01);PC三組肝臟ADA,XOD mRNA表達(dá)明顯下降,腎臟GLUT9,URAT1mRNA表達(dá)明顯下降,OAT1、OCT1 mRNA表達(dá)明顯上升(P0.01,P0.05);PC三組肝臟XOD蛋白表達(dá)明顯下降,腎臟GLUT9、URAT1蛋白表達(dá)明顯下降(P0.01,P0.05);PC中組小腸MRP4蛋白表達(dá)明顯升高(P0.01);腎臟組織病理切片表明:PC中組的大多數(shù)腎皮質(zhì)腎小管上皮細(xì)胞形態(tài)正常,胞漿豐富,腎髓質(zhì)少數(shù)集合管管腔內(nèi)見紅細(xì)胞堆積。結(jié)論:虎桂藥對成分——虎杖苷和桂皮醛相互配伍后,具有明顯的抗高尿酸血癥的藥理作用。可通過抑制尿酸的生成和促進(jìn)尿酸的排泄雙重降低血尿酸含量,改善高尿酸血癥對腎臟造成的器質(zhì)及功能性損傷。其作用機(jī)制可能與抑制大鼠肝臟XOD、ADA和腎臟URAT1、GLUT9的表達(dá);促進(jìn)腎臟OCT1、OAT1和小腸MRP4的表達(dá)相關(guān)。
[Abstract]:Hyperuricemia is one of the many metabolic diseases that perplex human beings. A large number of domestic and foreign studies have shown that high uric acid blood is closely related to many diseases, such as acute gouty arthritis, uric acid kidney disease, hypertension, obesity, diabetes and cardiovascular diseases. There has been a great change. The night, the fast food culture, the beer, the drink (including fructose) and so on in our country, caused the outbreak of hyperuricemia in our country to increase, and present a low age trend. It seriously affects the health of our people and the quality of life. At present, the research of hyperuricemia is still lack of recognition. In this experiment, we hope to try to establish a model of hyperuricemia in rats with a relatively gentle, stable and persistent blood uric acid value, and to explore the therapeutic effect of the combination of Polygonum cuspidin and cinnamaldehyde on hyperuricemia on the basis of this model. And its mechanism of action mechanism.1. rats hyperuricemia model experiment to optimize the dosage of hypoxanthine, potassium oxazine and ethambutol to prepare the model of hyperuricemia in rats, in order to get a more suitable model of hyperuricemia in rats. Methods: 32 male rats were randomly divided into 4 groups, 8 rats in each group. Group I: group I: group I: group I: group I: group I: group I: group I: group I: group I: group I: group I: group I: group I 75mg/kg hypoxanthine and ethambutol 250mg/kg were injected subcutaneously with potassium 200mg/kg oxazine (1 /d). Model II group was given 75mg/kg hypoxanthine and 250mg/kg hydrochloric ethambutol (1 /d), and hypodermic potassium oxazine (1 /12h). Model III group: gavage of 50mg/kg hypoxanthine and 250mg/kg hydrochloric ethambutol, 200mg/kg potassium oxazine (1 times /d) was injected subcutaneously in the blank group, and the normal saline was given to the stomach and the olive oil was injected subcutaneously for 24 days. After the subcutaneous injection of 4,11,18d, the blood was taken from the orbit of the rat and the content of SUA was measured. After twenty-fourth days of subcutaneous injection, the content of SUA in the serum was measured by 3,6,12,24h, and the content of BUN and Cr in the serum of 6h was measured. Rats were executed and the kidneys were killed and detected by the rats. Pathological changes. Results: compared with the blank group, the blood uric acid content in the three groups could increase significantly (P0.01), and could maintain 12h, especially in the model II group. The pathological section of the kidney of the three model rats showed that the three modes of modeling could lead to slight renal injury; the hematuria in model I group and model II Group The content of vegetarian nitrogen was significantly higher than that in the blank group (P0.01, P0.05). Conclusion: the model of hyperuricemia in rats can be successfully prepared by the three models, with slight renal organic and functional damage (especially in model III group), which has the advantage of long duration of increasing blood uric acid value (especially in model II Group sustainable 24h).2. The purpose of the treatment and mechanism of group distribution of tiger and cinnamon to hyperuricemia is to explore the effect of Polydatin and Cinnamaldehyde on the components of the Chinese medicine, and to explore the effect of uric acid on hyperuricemia in rats, and the excretion and formation of uric acid in two ways. To provide new ideas and methods for the treatment of hyperuricemia in Chinese medicine. Methods: 70 male SD rats were randomly divided into 7 groups. The blank group, the model group, the allopurinol group, the benzene bromide Malone group, the Polygonum glycoside and the cinnamaldehyde (PC) were low, middle, high group, 10 in each group. The subcutaneous injection of potassium oxazine (200mg/kg), 7h after 7h and hypoxanthine (250mg/kg) and hypoxanthine (250mg/kg). 50mg/kg) the rat model of hyperuricemia was prepared for 26 days (1 times /d). Fourth days after fourth days, the SUA value of rats was measured by 6h orbital injection after subcutaneous injection. From the fifth day, before subcutaneous injection, 1H, gavage for 21 days (1 times /d). The rats were put into the metabolic cage for twenty-fifth days, and the urine 24h was collected. After the last day subcutaneous injection of 6h, the abdominal aorta of rat was taken blood, measured blood, measured the abdominal aorta of rats, measured blood of rat abdominal aorta after the last day, measured blood of abdominal aorta of rats, measured blood of rat abdominal aorta, measured blood, measured abdominal aorta rats, measured blood, measured abdominal aorta rats, measured blood, measured abdominal aorta rats, measured blood, measured rat abdominal aorta, measured blood, measured rat abdominal aorta after blood, measured blood, measured rat abdominal aorta, measured blood after abdominal aorta, measured rat abdominal aorta after blood extraction, measured the rat aorta, test Determine the content of SUA, Cr, BUN and GPT; take out the.ELISA kit of rat liver, kidney and small intestine to determine the content of Kim-1 and NGAL in urine; the pathological observation of kidney tissue, statistical score; qPCR determination of liver XOD, ADA and kidney GLUT9, URAT1, protein expression; immunization The protein expression of MRP4 in the small intestine was determined by histochemistry and the cumulative photo density value (IOD) of the photo positive was analyzed. Results: after the model, the content of SUA in the rats increased significantly (P0.01). After the treatment, the content of SUA in the high group decreased significantly (P0.01, P0.05) in PC; the BUN content in the PC group was obviously lower (P0.05), and the content of the PC three groups decreased significantly. The expression of Kim-1 was significantly decreased (P0.01), the expression of ADA, XOD mRNA in PC three groups was significantly decreased, the expression of GLUT9 and URAT1mRNA in the kidneys was significantly decreased, OAT1 and OCT1 mRNA increased (P0.01,), and the expression of egg white in the three groups was obviously decreased. The histopathological sections of renal tissue showed that most of the renal tubular epithelial cells in group PC were normal, rich in cytoplasm and accumulated in a small collection of tubules in the renal medulla. Conclusion: after compatibility of Polygonum cuspidin and cinnamaldehyde, the pharmacology of tiger cinnamon and Polygonum cuspidin and cinnamaldehyde have obvious pharmacological effects on anti hyperuricemia. Inhibiting the formation of uric acid and promoting uric acid excretion to reduce the content of uric acid and improve the organic and functional damage caused by hyperuricemia on the kidney. The mechanism may be related to the inhibition of the expression of XOD, ADA, URAT1 and GLUT9 in the rat liver, and the expression of OCT1, OAT1 and the expression of MRP4 in the small intestine.

【學(xué)位授予單位】：廣東藥科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R285.5

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