發(fā)布時(shí)間：2018-07-18 21:29

【摘要】：目的:商陸屬于峻下逐水藥,藥理作用較為廣泛,本研究旨在更深入的探索其抗腎炎作用,篩選出對(duì)雞腎腫癥治療效果最佳的提取方法,為臨床指導(dǎo)用藥和研制開(kāi)發(fā)新藥提供科學(xué)依據(jù)。方法:(1)采用水煎法、水提醇沉法(醇沉終濃度分別為60%、70%、80%、90%,)、減壓干燥法、滲漉法(乙醇濃度分別為50%、70%、90%)4種提取方法對(duì)商陸中的有效成分進(jìn)行提取,主要比較各提取方法中商陸皂苷甲的含量。(2)制造雞腎腫模型,然后用各商陸提取物進(jìn)行治療,并設(shè)立獸藥對(duì)照組和陽(yáng)性對(duì)照組,觀察治療前后各組雞的臨床癥狀、體重變化、治愈情況、臟器指數(shù)變化、血清中尿酸、肌酐、尿素、腎組織NO含量、白細(xì)胞介素2受體水平和Caspase-3活性。(3)對(duì)商陸滲漉提取物治療雞腎腫癥的療效進(jìn)行安全性評(píng)價(jià),試驗(yàn)動(dòng)物分5組,分別為正常劑量組、正常劑量3倍、5倍、10倍量組和空白對(duì)照組,觀察試驗(yàn)動(dòng)物在用藥期間臨床癥狀、血液學(xué)指標(biāo)、血清生化指標(biāo)及組織切片與空白對(duì)照組相比有無(wú)明顯差異。(4)對(duì)商陸皂苷甲含量測(cè)定和薄層鑒別的專(zhuān)屬性、重復(fù)性等有關(guān)方法學(xué)試驗(yàn)進(jìn)行研究。結(jié)果:(1)試驗(yàn)結(jié)果顯示用10倍量90%的乙醇浸提12 h,滲漉速度為1mL/min,得到的商陸皂苷甲的含量和提取率最高,分別為1.72 mg/mL和49.14%。(2)臨床藥效學(xué)研究結(jié)果顯示:不同方法提取得到的商陸提取物對(duì)雞腎腫癥均具有顯著的改善和治療效果,其中商陸滲漉提取物治療效果最佳,能顯著降低模型組雞只血清中尿酸、肌酐、尿素、腎組織NO、白細(xì)胞介素2受體水平,和空白對(duì)照組相比差異不顯著(P0.05),各模型用藥組之間與空白對(duì)照組中Caspase-3活性均無(wú)顯著差異(P0.05)。(3)商陸滲漉提取物治療雞腎腫癥安全性評(píng)價(jià)試驗(yàn)結(jié)果顯示:用藥期間各用藥組的臨床癥狀無(wú)明顯變化,商陸滲漉提取物最高劑量組體重在用藥期結(jié)束時(shí)低于空白對(duì)照組,但差異不顯著(P0.05),各用藥組血液學(xué)指標(biāo)和血清生化指標(biāo)與空白對(duì)照組比較無(wú)顯著差異,剖檢和組織學(xué)觀察未見(jiàn)明顯異常。(4)質(zhì)量研究試驗(yàn)結(jié)果顯示,建立的商陸皂苷甲檢測(cè)方法專(zhuān)屬性強(qiáng)、重復(fù)性高。結(jié)論:研究表明用10倍量90%的乙醇浸提12 h,滲漉速度為1 mL/min,得到的商陸皂苷甲的含量和提取率最高,分別為1.72 mg/mL和49.14%;且在藥效學(xué)研究中,對(duì)腎臟腫大的改善和治療效果最為突出,推測(cè)商陸皂苷甲可能是治療腎腫癥的有效成分之一;在安全性評(píng)價(jià)中各用藥組血液學(xué)指標(biāo)和血清生化指標(biāo)與空白對(duì)照組比較無(wú)顯著差異,剖檢和組織學(xué)觀察未見(jiàn)明顯異常,說(shuō)明商陸提取物用于治療雞的腎臟疾病是安全有效的;最后分別建立了商陸皂苷甲的含量測(cè)定和薄層鑒別方法,且方法簡(jiǎn)便易操作,穩(wěn)定可靠,為更好的控制質(zhì)量提供依據(jù)。
[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.
【學(xué)位授予單位】：河南科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：S858.31

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