發(fā)布時間：2018-06-18 16:58

  本文選題：犬流感病毒 + 苦參　； 參考：《山東農業(yè)大學》2015年碩士論文

【摘要】：犬流感是由犬流感病毒(Canine influenza virus,CIV)引起的以體溫升高、咳嗽、呼吸困難為特征的犬的急性傳染性疾病,其特點是傳播快,流行廣。我國和韓國都在呼吸道嚴重感染犬中分離到H3N2亞型流感病毒,實驗發(fā)現這種病毒可以在犬之間進行直接傳播。在現代生活中,犬是人類的忠實朋友,近年來寵物犬數量急劇增加,這也為犬流感病毒的跨宿主傳播提供了更多的機會,人們需要重視這一問題。西藥在抗犬流感病毒的臨床應用上尚存在極大的局限性,如降低畜禽免疫力,易產生抗藥性、具有副作用等。中藥毒副作用小、不易產生耐藥性,近年來的研究逐漸增多。但是把中藥的藥理活性、有效成分與藥物的基礎功效結合起來進行綜合性研究的報道仍較少見。本課題基于前期研究成果即中藥苦參具有良好的體外抗犬流感病毒效果的結論,以苦參為原材料,進行苦參主要成分的抗流感病毒效果研究。本試驗采用稀醇提取、大孔樹脂純化法,從苦參中獲得了主要成分即生物堿類,黃酮類,皂苷類和多糖類化合物。用MDCK細胞建立體外實驗模型,將苦參提取物進行有效處理,制成可用于MDCK細胞的藥物制劑,以利巴韋林為陽性對照,應用細胞病變觀察法和MTT法檢測細胞活性,對苦參成分的抗犬流感病毒效果進行評價。首先測定各成分對MDCK的最大無毒濃度,然后在安全濃度范圍內比較四種成分的抗犬流感病毒效果,通過改變給藥時間及作用方式初步探討有效成分抗犬流感病毒的作用環(huán)節(jié),最后確定其最小有效濃度。結果表明:苦參四種主要成分的最大安全濃度在3.344μg/m L~1.229 mg/m L之間。四種主要成分均有顯著的抗CIV效果(P0.05)且綜合作用效果均優(yōu)于利巴韋林。不同給藥方式試驗結果顯示:(1)苦參四種成分均能阻止CIV吸附進入細胞,且效果均顯著優(yōu)于利巴韋林(P0.05),生物堿和皂苷類的效果優(yōu)于其他兩種成分;(2)苦參四種成分均能干擾CIV的合成與釋放;(3)病毒和藥物同時接觸細胞時,苦參四種成分均能起到抑制CIV的作用,生物堿類與利巴韋林作用相近,但黃酮、皂苷、多糖類成分的效果顯著低于利巴韋林(P0.05);(4)藥物與病毒預先作用時,苦參四種成分均能起到顯著的殺滅CIV的作用,皂類與利巴韋林的作用相近,但生物堿、黃酮、多糖成分的效果顯著低于利巴韋林(P0.05)�？鄥⒂行С煞值淖钚∮行舛仍�0.2090μg/mL~0.0474 mg/m L之間。不同成分的治療指數(TI)在16~128之間,其中生物堿和多糖的治療指數最高,為128,皂苷的治療指數為32,黃酮的治療指數為16。
[Abstract]:Canine influenza is an acute infectious disease caused by canine influenza virus (influenza), which is characterized by elevated body temperature, cough and dyspnea. It is characterized by rapid transmission and widespread prevalence. H3N2 subtype influenza virus was isolated from dogs with severe respiratory tract infection in China and South Korea, and it was found that H3N2 subtype influenza virus can be transmitted directly from dog to dog. In modern life, dogs are the faithful friends of human beings. In recent years, the number of pet dogs has increased dramatically, which also provides more opportunities for the cross-host transmission of canine influenza virus. People should pay attention to this problem. Western medicine has great limitations in the clinical application of canine influenza virus, such as reducing the immunity of livestock and poultry, easily producing drug resistance, and having side effects, etc. The research of traditional Chinese medicine is increasing in recent years because of its low toxicity and resistance. However, the comprehensive study of pharmacological activity, active components and basic efficacy of traditional Chinese medicine is still rare. Based on the conclusion that Sophora flavescens has good antiviral effect in vitro, this paper studies the antiviral effect of Sophora flavescens as raw material. In this experiment, alkaloids, flavonoids, saponins and polysaccharides were obtained from Sophora flavescens by extraction of dilute alcohol and purification of macroporous resin. The experimental model of MDCK cells was established in vitro. The extract of Sophora flavescens was effectively treated to prepare the drug preparation for MDCK cells. Ribavirin was used as positive control. The cytopathic observation method and MTT method were used to detect the cell activity. The effect of Sophora flavescens against canine influenza virus was evaluated. First, the maximum nontoxic concentration of each component to MDCK was determined, and then the anti-canine influenza virus effect of four ingredients was compared within the safe concentration range, and the effect of the active ingredient on the canine influenza virus was preliminarily discussed by changing the administration time and the mode of action. Finally, the minimum effective concentration was determined. The results showed that the maximum safe concentration of four main components of Sophora flavescens was between 3.344 渭 g / mL and 1.229 mg/m / L. The four main components had significant anti-CIV effect (P0.05) and the comprehensive effect was superior to ribavirin. The results of different drug administration methods showed that all four components of Sophora flavescens could prevent CIV from adsorbing into cells. The effects were significantly better than that of ribavirin P0.05, alkaloids and saponins were superior to those of the other two components. All four components of Sophora flavescens could interfere with the synthesis and release of CIV, and when the virus and drugs were in contact with the cells at the same time, the four components of Sophora flavescens could interfere with the synthesis and release of CIV. The effects of alkaloids and ribavirin on CIV were similar, but the effects of flavonoids, saponins and polysaccharides were significantly lower than those of ribavirin (P0.05). The effects of soap and ribavirin were similar, but the effects of alkaloids, flavonoids and polysaccharides were significantly lower than that of ribavirin. The minimum effective concentration of active components of Sophora flavescens was 0.2090 渭 g / mL 0.0474 mg/m / L. Among them, alkaloids and polysaccharides had the highest treatment index (128), saponins (32) and flavonoids (16).
【學位授予單位】：山東農業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S853.7

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本文編號：2036184