本文關(guān)鍵詞：脫氫醋酸鈉致大鼠和家兔出血的初步研究　出處：《揚(yáng)州大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 脫氫醋酸鈉 出血 PT APTT VK

【摘要】：目的：脫氫醋酸鈉(Sodium Dehydroacetate)抗菌譜廣,具有較好的防霉效果,作為防腐劑廣泛用于食品、飼料及化妝品等相關(guān)領(lǐng)域。高劑量應(yīng)用脫氫醋酸鈉可能引起大鼠有出血傾向。本課題組前期研究中也發(fā)現(xiàn)脫氫醋酸鈉能引起大鼠多器官出血。本文通過(guò)給予大鼠和家兔不同劑量脫氫醋酸鈉,檢測(cè)凝血指標(biāo)PT和APTT,測(cè)定血液中VK.1、脫氫醋酸鈉血藥濃度及VKORC1蛋白的表達(dá),并對(duì)主要臟器進(jìn)行HE染色組織病理學(xué)分析,研究脫氫醋酸鈉應(yīng)用于大鼠及家兔后引起的出血傾向。方法：Wistar大鼠,雌雄各半,分別灌胃給予脫氫醋酸鈉50mg/kg、100mg/kg、150 mg/kg和200 mg/kg,生理鹽水作為對(duì)照組,每組14只。每天一次,連續(xù)13d。家兔灌胃給予脫氫醋酸鈉分為100 mg/kg組、200 mg/kg組和生理鹽水對(duì)照組,每組14只。在給藥后第3d、5d、7d、9d、11d和13d分別采集血液和主要臟器,每組大鼠或家兔4只。測(cè)定PT和APTT,對(duì)肝臟、脾臟、肺臟、心臟和腎臟進(jìn)行組織病理學(xué)檢查,HPLC法檢測(cè)血液中的脫氫醋酸鈉濃度,ELISA法檢測(cè)大鼠血清VK1含量,免疫組化檢測(cè)組織中VKORC1蛋白表達(dá)。HPLC法檢測(cè)血清中脫氫醋酸鈉的濃度,色譜條件為X BridgeC18色譜柱,甲醇-0.02 mol/L乙酸銨(35/65,v/v)為流動(dòng)相,293 nm檢測(cè)。血液樣品經(jīng)乙腈提取、濃縮后上機(jī)測(cè)定。結(jié)果：不同劑量脫氫醋酸鈉給予大鼠后,與對(duì)照組比較PT和APTT值極顯著延長(zhǎng)(P0.001),且雄鼠的PT和APTT值顯著高于雌鼠(P0.05)。對(duì)大鼠HE染色組織切片分析,200 mg/kg組心臟、肝臟、脾臟、肺臟和腎臟都有明顯的出血；肝臟組織的出血程度與脫氫醋酸鈉劑量有關(guān)。100-200 mg/kg劑量脫氫醋酸鈉均可引起血清中VK1的含量顯著低于生理鹽水對(duì)照組(P0.01)。 VKORC1蛋白在大鼠肝臟、肺臟、脾臟、心臟和腎臟的表達(dá)水平均顯著低于對(duì)照組(P0.01)。 HPLC法測(cè)定大鼠血清中脫氫醋酸鈉的濃度在20-50 mg/L。相關(guān)性分析表明大鼠血清中VK1水平與PT和APTT的相關(guān)系數(shù)分別為0.719和0.599；大鼠血清中脫氫醋酸鈉濃度與PT和APTT的相關(guān)系數(shù)分別為0.900-0.988和0.806-0.852。與對(duì)照組比較,100-200 mg/kg劑量脫氫醋酸鈉給予家兔7d后,PT和APTT值極顯著地延長(zhǎng)(P0.001)；組織病理學(xué)分析200 mg/kg組除心臟外,肝臟,脾臟,肺臟和腎臟都有明顯的出血。HPLC法測(cè)定脫氫醋酸鈉在家兔的血藥濃度為5-20 mg/L。相關(guān)性分析表明家兔血清中脫氫醋酸鈉血藥濃度與PT和APTT的相關(guān)系數(shù)分別為0.837和0.351。結(jié)論：較高劑量(150-200 mg/kg)脫氫醋酸鈉可引起大鼠和兔凝血指標(biāo)明顯延長(zhǎng),主要臟器有明顯的出血；引起大鼠血液中VK1水平明顯降低,組織中VKORC1表達(dá)降低。脫氫醋酸鈉致大鼠或家兔PT和APTT延長(zhǎng),與脫氫醋酸鈉血藥濃度以及血清中VK1水平高度相關(guān)。脫氫醋酸鈉導(dǎo)致大鼠和家兔的出血,可能與抑制VKORC1,影響VK循環(huán),導(dǎo)致血清中VK1的水平降低有關(guān)。
[Abstract]:Objective: sodium Dehydroacetate has a wide antibacterial spectrum and a good anti-mildew effect. It is widely used as a preservative in food. High dose of sodium dehydroacetate may cause bleeding tendency in rats. We also found that sodium dehydroacetate can cause multiple organ bleeding in rats. And different doses of sodium dehydroacetate in rabbits. Blood coagulation parameters PT and APTT, blood VK.1, blood concentration of sodium dehydroacetate and expression of VKORC1 protein were detected. The histopathological analysis of main organs was performed by HE staining. To study the bleeding tendency induced by sodium dehydroacetate in rats and rabbits. Methods Wistar rats, male and female, were given 50 mg / kg of sodium dehydroacetate and 100 mg / kg of sodium dehydroacetate respectively. 150 mg/kg and 200 mg / kg, normal saline as control group, 14 rats in each group, once a day. For 13 consecutive days, the rabbits were given sodium dehydroacetate intragastrically, which were divided into two groups: 100 mg/kg group (n = 14) and saline control group (n = 14). At the 3rd day, 5 days after administration, the rabbits received sodium dehydroacetate for 7 days. Blood and main organs were collected at 9 d and 13 d, respectively. PT and APTT were measured in each group of rats or rabbits. Histopathological examination of liver, spleen, lung, heart and kidney were carried out. HPLC method was used to detect the concentration of sodium dehydroacetate in blood. Elisa was used to detect the content of serum VK1 in rats. The expression of VKORC1 protein in tissues was detected by immunohistochemistry. The concentration of sodium dehydroacetate in serum was detected by HPLC. The chromatographic condition was X BridgeC18 column. Methanol-0.02 mol/L ammonium acetate 35 / 65 v / v / v) was detected by mobile phase at 293nm. Blood samples were extracted by acetonitrile. Results: after different doses of sodium dehydroacetate were given to rats, PT and APTT values were significantly longer than those of control group (P 0.001). The PT and APTT values of male rats were significantly higher than that of female rats (P 0.05). The heart, liver and spleen of 200 mg/kg group were analyzed by HE staining. There were obvious bleeding in lung and kidney. The degree of hemorrhage in liver tissue was related to the dosage of sodium dehydroacetate. 100-200 mg/kg of sodium dehydroacetate could induce the content of VK1 in serum significantly lower than that in the control group (P < 0.05). VKORC1 protein was found in rat liver. Lungs, spleen. The expression levels in heart and kidney were significantly lower than those in control group (P 0.01). The concentration of sodium dehydroacetate in serum of rats was determined by HPLC method in the range of 20-50. The correlation analysis of mg / L showed that the correlation coefficients between VK1 level and PT and APTT were 0.719 and 0.599 respectively. The correlation coefficients between the concentration of sodium dehydroacetate and PT and APTT in serum of rats were 0.900-0.988 and 0.806-0.852.Compared with the control group. The values of PT and APTT in rabbits treated with sodium dehydroacetate for 100-200 mg/kg for 7 days were significantly prolonged (P 0.001). Histopathological analysis was performed in the 200 mg/kg group except the heart, liver and spleen. The blood concentration of sodium dehydroacetate in rabbits was 5-20 by HPLC. Correlation analysis showed that the correlation coefficients between serum concentration of sodium dehydroacetate and PT and APTT in rabbit serum were 0.837 and 0.351 respectively. Sodium dehydroacetate (150 ~ 200 mg / kg) significantly prolonged the coagulation parameters of rats and rabbits. There was obvious bleeding in the main organs. The levels of VK1 in blood and the expression of VKORC1 in tissues were significantly decreased, and PT and APTT were prolonged in rats or rabbits induced by sodium dehydroacetate. It was highly correlated with the concentration of sodium dehydroacetate in blood and the level of VK1 in serum. Sodium dehydroacetate caused hemorrhage in rats and rabbits, which may be related to the inhibition of VKORC1 and the influence of VK circulation. It is related to the decrease of serum VK1 level.
【學(xué)位授予單位】：揚(yáng)州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：S852.3

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