發(fā)布時間：2018-07-28 06:35

【摘要】：目的:分析準噶爾烏頭乙醇提取物的化學成分,觀察其單次灌胃后24h內(nèi)對大鼠血液生化指標及其內(nèi)源性代謝物的影響,為進一步研究該藥的毒性機理、臨床中毒診斷及新藥開發(fā)提供科學依據(jù)。方法:采用高效液相色譜(HPLC)技術(shù)對準噶爾烏頭乙醇提取物的化學成分進行定性定量分析;于大鼠單次灌胃準噶爾烏頭乙醇提取物(7.8mg/kg)前及灌胃后0.5、1、2、6、12、24h分別從大鼠眼眶后靜脈采集血液樣本,通過全自動生化分析儀檢測AST、ALT、TP、ALB、TC、TG、TBIL、Crea、GLU、LDH含量,用1H-NMR技術(shù)檢測大鼠血清內(nèi)源性代謝物的變化情況,分析給藥前后各參數(shù)值的變化;結(jié)合SIMCA軟件對以上數(shù)據(jù)進行模式識別(主成分分析法(PCA)、偏最小二乘法(PLS-DA)和正交偏最小二乘法-判別分析法(OPLS-DA)),尋找差異代謝物。結(jié)果:(1)準噶爾烏頭乙醇提取物HPLC譜圖的色譜條件為:乙腈-水(0.3%磷酸)(B:A)作為流動相,梯度洗脫:10~16%B(0~12 min)、16~35%B(12~30 min)、35~45%B(30~40 min)、45%B(40~45 min),檢測波長為235 nm,溫度35℃,流速1.0 mL/min,進樣量20μL。本方法在分離度、精密度、穩(wěn)定性、加樣回收率等方法學考察中均符合要求。(2)準噶爾烏頭乙醇提取物中含有宋果靈(54.9±4.5 mg)、苯甲酰烏頭原堿(7.3±1.6 mg)、烏頭堿(3.7±0.8 mg)、草烏甲素(1.2±0.3 mg)、3-脫氧烏頭堿(3.6±0.5 mg)共5種生物堿成分。(3)大鼠單次灌胃準噶爾烏頭乙醇提取物(7.8mg/kg)后起AST、TG、GLU、LDH升高(P0.05),ALB、TP、TC、TBIL降低(P0.05))。其中多數(shù)生化指標值在灌胃后1~2h內(nèi)出現(xiàn)變化高峰(升高或下降),對LDH和AST的影響最為顯著。(4)核磁共振氫譜(1H-NMR)代謝組學分析表明準噶爾烏頭乙醇提取物灌胃前后大鼠血清的1H-NMR譜存在明顯差異,主要差異性標記物有:α-葡萄糖、β-葡萄糖、肌酸水平升高(p0.05),氨基酸(丙氨酸、谷氨酸、甘氨酸、酪氨酸、亮氨酸、蘇氨酸、賴氨酸、苯丙氨酸)、脂類水平呈先降后升趨勢;丙酮酸和乳酸水平降低(p0.05)。結(jié)論:(1)建立的HPLC檢測方法穩(wěn)定可靠;(2)準噶爾烏頭乙醇提取物中含有宋果靈、苯甲酰烏頭原堿、烏頭堿、草烏甲素、3-脫氧烏頭堿5種生物堿成分。(3)準噶爾烏頭乙醇提取物可能對心肌、肝臟有不同程度的損害,并對大鼠的糖代謝、脂類代謝和氨基酸代謝產(chǎn)生影響。(4)大鼠內(nèi)源性代謝物的差異性變化有可能作為該藥中毒檢測的毒性標記物。
[Abstract]:Objective: to analyze the chemical constituents of the ethanol extract from Junggar aconitum and to observe the effect of the extract on the blood biochemical index and its endogenous metabolites in rats within 24 hours after single oral administration, in order to further study the toxic mechanism of the drug. The diagnosis of clinical poisoning and the development of new drugs provide scientific basis. Methods: the chemical constituents of ethanol extract of Junggar aconitum were qualitatively and quantitatively analyzed by high performance liquid chromatography (HPLC). Blood samples were collected from the posterior orbital vein of rats before and after single gastric administration of ethanol extract of Junggar aconitum (7.8mg/kg) and 0.5g / 2h after oral administration for 24 hours. The content of TBILTILTBILTILTBILLLU LDH was measured by automatic biochemical analyzer, and the results showed that the blood samples were collected from the posterior orbital vein of the rats before and after oral administration of the ethanol extract of Junggar aconitum (7.8mg/kg), and the contents of the blood samples were measured by automatic biochemical analyzer. The changes of serum endogenous metabolites in rats were detected by 1H-NMR technique, and the changes of parameters before and after administration were analyzed. The above data were identified with SIMCA software (principal component analysis, (PCA), partial least squares (PLS-DA) and orthogonal partial least squares discriminant analysis (OPLS-DA),) to search for different metabolites. Results: (1) the chromatographic conditions of HPLC spectra of ethanol extract of Zhunggar aconitum were as follows: acetonitrile-water (0.3% phosphoric acid) (B: a) as mobile phase, gradient elution of 10: 1016 and B (0 ~ 12 min) and B (12 ~ 30 min) ~ 35 ~ (35) B (3040 min) ~ 45B (4045 min), the detection wavelength was 235 nm, the temperature was 35 鈩，

本文編號：2149230