商陆活性成分的提取及其对鸡肾肿症治疗效果的研究

发布时间：2018-07-18 21:29

【摘要】：目的:商陆属于峻下逐水药,药理作用较为广泛,本研究旨在更深入的探索其抗肾炎作用,筛选出对鸡肾肿症治疗效果最佳的提取方法,为临床指导用药和研制开发新药提供科学依据。方法:(1)采用水煎法、水提醇沉法(醇沉终浓度分别为60%、70%、80%、90%,)、减压干燥法、渗漉法(乙醇浓度分别为50%、70%、90%)4种提取方法对商陆中的有效成分进行提取,主要比较各提取方法中商陆皂苷甲的含量。(2)制造鸡肾肿模型,然后用各商陆提取物进行治疗,并设立兽药对照组和阳性对照组,观察治疗前后各组鸡的临床症状、体重变化、治愈情况、脏器指数变化、血清中尿酸、肌酐、尿素、肾组织NO含量、白细胞介素2受体水平和Caspase-3活性。(3)对商陆渗漉提取物治疗鸡肾肿症的疗效进行安全性评价,试验动物分5组,分别为正常剂量组、正常剂量3倍、5倍、10倍量组和空白对照组,观察试验动物在用药期间临床症状、血液学指标、血清生化指标及组织切片与空白对照组相比有无明显差异。(4)对商陆皂苷甲含量测定和薄层鉴别的专属性、重复性等有关方法学试验进行研究。结果:(1)试验结果显示用10倍量90%的乙醇浸提12 h,渗漉速度为1mL/min,得到的商陆皂苷甲的含量和提取率最高,分别为1.72 mg/mL和49.14%。(2)临床药效学研究结果显示:不同方法提取得到的商陆提取物对鸡肾肿症均具有显著的改善和治疗效果,其中商陆渗漉提取物治疗效果最佳,能显著降低模型组鸡只血清中尿酸、肌酐、尿素、肾组织NO、白细胞介素2受体水平,和空白对照组相比差异不显著(P0.05),各模型用药组之间与空白对照组中Caspase-3活性均无显著差异(P0.05)。(3)商陆渗漉提取物治疗鸡肾肿症安全性评价试验结果显示:用药期间各用药组的临床症状无明显变化,商陆渗漉提取物最高剂量组体重在用药期结束时低于空白对照组,但差异不显著(P0.05),各用药组血液学指标和血清生化指标与空白对照组比较无显著差异,剖检和组织学观察未见明显异常。(4)质量研究试验结果显示,建立的商陆皂苷甲检测方法专属性强、重复性高。结论:研究表明用10倍量90%的乙醇浸提12 h,渗漉速度为1 mL/min,得到的商陆皂苷甲的含量和提取率最高,分别为1.72 mg/mL和49.14%;且在药效学研究中,对肾脏肿大的改善和治疗效果最为突出,推测商陆皂苷甲可能是治疗肾肿症的有效成分之一;在安全性评价中各用药组血液学指标和血清生化指标与空白对照组比较无显著差异,剖检和组织学观察未见明显异常,说明商陆提取物用于治疗鸡的肾脏疾病是安全有效的;最后分别建立了商陆皂苷甲的含量测定和薄层鉴别方法,且方法简便易操作,稳定可靠,为更好的控制质量提供依据。
[Abstract]:Objective: pokeweed is a kind of pokeweed with extensive pharmacological effects. The aim of this study was to explore its anti-glomerulonephritis effect and to select the best method for the treatment of chicken kidney swelling. To provide scientific basis for clinical guidance and research and development of new drugs. Methods: (1) four methods were used to extract the effective components of Phytolacca japonicus by water decoction, water extraction and alcohol precipitation (60% final concentration of alcohol precipitation was 60%), decompression and drying method, percolation method (ethanol concentration was 50,707070%). The content of pokeweed saponins A was compared in each extraction method. (2) the chicken kidney swelling model was made, then treated with each pokeweed extract, and the veterinary drug control group and the positive control group were set up to observe the changes of clinical symptoms and body weight of each group before and after treatment. Cure condition, organ index change, serum uric acid, creatinine, urea, no content in renal tissue, interleukin 2 receptor level and Caspase-3 activity. (3) to evaluate the efficacy of pokeweed percolation extract in the treatment of chicken renal swelling. The experimental animals were divided into 5 groups: normal dose group, normal dose group and blank control group. The clinical symptoms and hematological indexes were observed. There were significant differences in serum biochemical indexes and tissue sections between the control group and the control group. (4) the specificity and reproducibility of the determination of pokeweed saponin A content and the identification of thin-layer were studied. Results: (1) the results showed that the content and extraction rate of pokeweed saponin A was the highest when 90% ethanol was used for 12 h and the percolation rate was 1 mL / min. (2) the results of clinical pharmacodynamics study showed that the extraction of pokeweed extract obtained by different methods had significant improvement and therapeutic effect on chicken nephropathy, among which the percolation extract of pokeweed had the best therapeutic effect. The levels of serum uric acid, creatinine, urea, no and interleukin 2 receptor in renal tissue were significantly decreased in model group. Compared with the blank control group, there was no significant difference in Caspase-3 activity between each model drug group and the blank control group (P0.05). (3). There was no significant change in clinical symptoms in the drug group. The body weight of the highest dose group of pokeweed percolation extract was lower than that of the blank control group at the end of the medication period, but the difference was not significant (P0.05). (4) the results of quality study showed that the established method for the detection of pokeweed saponin A had strong specificity and high reproducibility. Conclusion: the results showed that the content and extraction rate of pokeweed saponin A were 1.72 mg / mL and 49.14%, respectively, and the percolation rate was 1 mL / min with 10 times 90% ethanol for 12 h, and in the pharmacodynamics study, the content and extraction rate of pokeweed saponin A were 1.72 mg / mL and 49.14 mg / mL, respectively. It is suggested that pokeweed saponin A may be one of the effective components in the treatment of renal swelling. In the safety evaluation, there was no significant difference in hematological index and serum biochemical index between each drug group and the blank control group, and there was no obvious abnormality in dissection and histological observation, which indicated that pokeweed extract was safe and effective in the treatment of chicken kidney disease. Finally, the content determination and TLC identification method of pokeweed saponin A were established, and the method was simple, easy to operate, stable and reliable, which provided the basis for better quality control.
【学位授予单位】：河南科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S858.31

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