本文選題：板藍(lán)根多糖 + 肺氣虛證��； 參考：《河南農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：肺氣虛證導(dǎo)致肺氣虧損,體內(nèi)宗氣合成不足,宣發(fā)無力,進(jìn)而表現(xiàn)出免疫功能低下的狀態(tài)。板藍(lán)根是傳統(tǒng)清熱藥,主入肺經(jīng),具有清熱解毒、涼血利咽功效。多糖作為板藍(lán)根的主要活性成分,其良好的抗病毒和免疫調(diào)節(jié)能力受到眾多研究者的青睞。豬流感病毒是引起豬呼吸道傳染病的重要病原,H3N2型豬流感病毒在豬場廣泛存在,可破壞豬的免疫系統(tǒng)進(jìn)而引起免疫抑制,在規(guī)�；i場難以根除。目前,豬流感無特效藥,中藥因其資源豐富、毒副作用小、價(jià)格低廉等特點(diǎn)有著獨(dú)特的優(yōu)勢。本試驗(yàn)通過利用MDCK細(xì)胞模型和中醫(yī)癥候動物模型分別評價(jià)和觀察了板藍(lán)根多糖體外和體內(nèi)抗H3N2型豬流感病毒的作用,豐富了中藥抗流感病毒藥庫,篩選以中醫(yī)證候模型為基礎(chǔ)的抗病毒新藥,也為中醫(yī)證候?qū)W研究、中西醫(yī)結(jié)合防治動物疫病提供更多理論依據(jù)�，F(xiàn)將本文主要研究內(nèi)容及研究成果歸納如下:一、采用水煮醇沉法提取板藍(lán)根多糖,利用MDCK細(xì)胞體外培養(yǎng)系統(tǒng),通過MTT法檢測細(xì)胞活性評價(jià)板藍(lán)根多糖體外抑制A/Swine/Henan/2010(H3N2)豬流感病毒對細(xì)胞的感染作用。結(jié)果表明,板藍(lán)根多糖對MDCK細(xì)胞表現(xiàn)出一定的毒性作用,半數(shù)毒性濃度(TC50)為7.273mg/mL,最大安全濃度(TC0)為2.504mg/mL,板藍(lán)根多糖對豬流感病毒的阻斷、抑制和直接殺滅作用治療指數(shù)分別是10.158、19.291、0.038。板藍(lán)根多糖在MDCK細(xì)胞上對豬流感病毒的抑制作用和阻斷作用效果較好,在此藥物安全濃度范圍內(nèi),板藍(lán)根多糖直接殺滅作用不明顯。二、采用A/Swine/Henan/2010(H3N2)豬流感病毒經(jīng)鼻腔感染6-8周肺氣虛證BALB/c小鼠,通過觀察和分析感染后小鼠的體重,臟器指數(shù),病理學(xué)變化以及免疫學(xué)等相關(guān)指標(biāo),探討H3N2型豬流感病毒感染肺氣虛證BALB/c小鼠動物模型致病性變化。結(jié)果顯示,感染豬流感病毒肺氣虛證小鼠精神狀態(tài)表現(xiàn)較差,體重下降幅度大,肺臟出現(xiàn)水腫、出血和炎性細(xì)胞大量增生、細(xì)支氣管黏膜上皮嚴(yán)重脫落、血管周圍炎性細(xì)胞浸潤等病變,機(jī)體淋巴增值能力、NO、IgG水平降低,促炎因子和抗炎因子水平紊亂且通過抑制IFN-γ與TNF-α水平影響其間接抗病毒能力。此外,從體重、肺部病理變化和機(jī)體損傷程度等方面綜合分析,肺氣虛證小鼠加重了其感染豬流感病毒引起的病變和損傷。三、以豬流感病毒A/Swine/Henan/2010(H3N2)感染肺氣虛證BALB/c小鼠為模型,經(jīng)板藍(lán)根多糖作用后分別在第2,5,9天觀察和檢測其體征狀態(tài)、體重增長率、臟器指數(shù)、肺臟病理變化、腹腔巨噬細(xì)胞活性、淋巴細(xì)胞增殖能力、NO和IgG水平變化、細(xì)胞因子等免疫學(xué)水平。結(jié)果顯示,經(jīng)板藍(lán)根多糖干預(yù)后可明顯改善感染小鼠體征狀態(tài),提高體重增長率并提前恢復(fù)至感染前水平,減輕或減小了肺臟水腫、出血、炎性細(xì)胞浸潤、肺泡間質(zhì)增生和病變區(qū)域。板藍(lán)根多糖可能通過促進(jìn)小鼠促炎細(xì)胞因子如IL-1β、IL-6和TNF-α水平,調(diào)節(jié)免疫應(yīng)答,增強(qiáng)機(jī)體炎性修復(fù)能力;通過促進(jìn)IL-12和IFN-γ的分泌,增強(qiáng)機(jī)體抗病毒作用,減輕肺臟炎性反應(yīng)下調(diào)IL-10的表達(dá)。此外,不同劑量板藍(lán)根多糖在機(jī)體抵抗流感病毒生物學(xué)活性調(diào)節(jié)活動中所起的作用有所差異,其中綜合評價(jià)灌胃濃度75 mg·kg-1效果最好。綜上,豬流感病毒A/Swine/Henan/2010(H3N2)感染肺氣虛證BALB/c小鼠可引起機(jī)體一系列致病性變化,且肺氣虛證小鼠加重了其感染豬流感病毒引起的病變和損傷;板藍(lán)根多糖可在細(xì)胞水平和動物機(jī)體水平上發(fā)揮直接和間接抗豬流感病毒作用。
[Abstract]:The deficiency of the lung qi caused by deficiency of the lung qi, the inadequacy of the body gas synthesis, the weakness of the hair, and the low immune function. The Radix Isatis Root is a traditional antipyretic drug, which is mainly into the lung meridian, and has the effect of clearing heat and detoxifying, cooling blood and taking advantage of the pharynx. The polysaccharide is the main active ingredient of the Radix Isatidis, and its good antiviral and immune regulation ability is subject to many researchers. Swine influenza virus is an important pathogen causing respiratory infectious diseases in pigs. H3N2 swine influenza virus exists widely in pig farms. It can destroy the immune system of pigs and cause immunosuppression and can not be eradicated in large-scale pig farms. At present, there are no special drugs for swine flu. The traditional Chinese medicine has unique characteristics because of its rich resources, small toxic side effects and low price. In this experiment, the effect of MDCK cell model and TCM syndrome model was used to evaluate and observe the anti H3N2 swine influenza virus in vitro and in vivo of Radix Isatidis, enriching the anti influenza virus pharmacy store in Chinese medicine, screening new antiviral drugs based on TCM syndrome model, and also for TCM syndrome study and Chinese and Western medicine. The main research contents and research results of this paper are summarized as follows: first, the extraction and alcohol precipitation method is used to extract the polysaccharide of Radix Isatidis, and the cell activity of MDCK cells in vitro is used to detect the cell activity by MTT method to evaluate the inhibition of A/Swine/Henan/2010 (H3N2) virus to the cells in vitro. The results showed that the polysaccharide of Radix Isatidis showed a certain toxic effect on MDCK cells, half of the toxic concentration (TC50) was 7.273mg/mL, the maximum safety concentration (TC0) was 2.504mg/mL, the blocking, inhibition and direct killing effect of Isatis polysaccharide on swine influenza virus was 10.158,19.291,0.038. Isatis polysaccharide in MDCK thin, respectively. The inhibitory effect and blocking effect of the cell on the swine influenza virus was better. In the range of the drug safety, the direct killing effect of Radix Isatidis polysaccharide was not obvious. Two, the A/Swine/Henan/2010 (H3N2) swine influenza virus was used to observe and analyze the body weight and organ index of the mice after the infection of the BALB/c mice after the nasal infection for 6-8 weeks. The pathogenicity of H3N2 swine influenza virus infected BALB/c mice was investigated by pathological changes and immunology. The results showed that the mental state of the mice infected with swine influenza virus lung qi deficiency syndrome was poor, the weight loss was large, the lungs were swollen, bleeding and inflammatory cells proliferated, and the bronchioles were sticky. The membrane epithelium is severely shedding, perivascular inflammatory cell infiltration and other lesions, the body lymphatic increment ability, NO, IgG level, the level of proinflammatory and anti-inflammatory factors are disorganized and the ability to resist the anti viral activity between IFN- gamma and TNF- alpha is influenced by the comprehensive analysis of the body weight, the pathological changes of the lungs and the degree of injury of the body. The mice aggravated the disease and damage caused by the infection of the swine influenza virus. Three, the BALB/c mice infected with the lung qi deficiency syndrome of the swine influenza virus A/Swine/Henan/2010 (H3N2) were used to observe and detect the physical signs, the weight growth rate, the organ index, the pathological changes of the lungs and the peritoneal macrophages on day 2,5,9 after the action of the Radix Isatidis polysaccharides. The activity, lymphocyte proliferation, NO and IgG levels, cytokines and other immunological levels. The results showed that the signs of infection in mice can be significantly improved by the prognosis of Radix Isatidis polysaccharides, increase the rate of weight growth and advance to the pre infection level, reduce or reduce the pulmonary edema, bleeding, inflammatory cell infiltration, alveolar interstitial hyperplasia. The diseased area. The polysaccharide of the Radix Isatidis may be promoted by promoting the level of IL-1 beta, IL-6 and TNF- alpha in mice, regulating the immune response and enhancing the ability of inflammatory repair. By promoting the secretion of IL-12 and IFN- gamma, the antiviral effect of the body is enhanced and the expression of IL-10 under the inflammatory response of the lung is reduced. The effect of body resistance to the biological activity regulation of influenza virus was different, in which the comprehensive evaluation of the concentration of 75 mg. Kg-1 was best. To sum up, the swine influenza virus A/Swine/Henan/2010 (H3N2) infection of the lung qi deficiency syndrome BALB/c mice could cause a series of pathogenic changes in the body, and the lung qi deficiency mice aggravated the infection of the swine influenza. Virus induced lesions and injuries; isatis root polysaccharides can play a direct and indirect role against swine influenza virus at cellular level and animal level.

【學(xué)位授予單位】：河南農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S853.7

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