清热化湿透表法干预小鼠流感病毒性湿热证免疫损伤的机理研究
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摘要
研究目的
近年来,中医药在治疗病毒性疾病方面逐步体现出独特的优势,特别是中药复方多靶点作用,整体调节机体免疫,达到“祛邪不伤正”的目的。呼吸道病毒感染是岭南地区临床常见病、多发病。病毒学检测发现,成人以流感病毒、副流感病毒感染为主。本病的临床治疗采用辨病与辨证相结合,临证治疗相当灵活。常用的中医治法有清热化湿、清热解毒、益卫固表等,其作用机理与抑制病毒复制、调节机体免疫、减少局部炎症渗出等有关。上述研究加深了对运用中医药治疗呼吸道病毒感染的认识,但还存在不少亟待解决的问题。本病的发生与病毒、机体、环境等多种因素相关,不同治法在疾病的不同阶段发挥作用,如何在治法研究中体现“辨证”的思路,有待进一步探讨。
研究表明,岭南地区呼吸道病毒感染从中医辨证具有易化热挟湿的特点。本研究前期的临床调查亦表明急性上呼吸道感染发病初起即易挟湿。清热化湿透表法是该地区常用外感热病治法之一,其中用于气分湿热证的蒿芩清胆汤除了具有清热化湿作用外,方中青蒿更能透邪达表,是清热化湿透表的代表方之一。以该方为主的经验治法广泛应用于岭南地区的呼吸道病毒感染治疗,具有一定地域特点,体现了中医辨证“三因制宜”的思路。为进一步明确岭南地区呼吸道病毒感染湿热证的内涵和外延,探讨清热化湿透表法的作用机理,我们进行了以下研究,以期为临床辨证运用清热化湿透表法治疗呼吸道病毒感染提供理论、临床和实验研究依据。
研究方法
本研究通过文献回顾及对101例急性上呼吸道感染患者的证候特点及用药特点的观察,在传统湿热证动物模型基础上,采用“肥甘饮食+湿热环境+流感病毒感染”方法,构建小鼠流感病毒性肺炎湿热证模型。以蒿芩清胆汤代表清热化湿透表法,以肺指数初步评价该法的治疗效果,以流式细胞术检测模型小鼠T淋巴细胞CD_3~+/CD_4~+,CD_8~+T亚群变化,以ELISA法检测血清IFN-γ及IL-4水平,通过Th1/Th2类细胞平衡变化评估机体免疫功能,采用免疫组织化学法检测肺组织炎症因子NF-κB及水通道蛋白AQP4的阳性表达,探讨清热化湿透表法干预病毒性肺炎湿热证的作用方向,并与银翘散(辛凉解表法)、玉屏风散(益卫固表法)比较异同。
研究结果及分析
模型构建方面:①造模成功后,模型小鼠体重减轻、肛温升高,蜷缩懒动、反应迟钝、呼吸急促甚则口唇紫绀,肺组织发生炎性实变。模型鼠症状的出现较单纯病毒感染小鼠滞后,似有减轻趋势。②模型小鼠感染病毒后,T淋巴细胞亚群比例失调,CD_4~+T%降低,CD_8~+T%升高,CD_4~+/CD_8~+比例倒置,病毒感染造成小鼠免疫系统抑制;致炎因子IFN-γ分泌显著增加,而IL-4分泌水平未见明显变化,Th1/Th2细胞因子平衡向Th1方向偏移,与病毒组相比差异不显著。③模型小鼠肺泡壁细胞核因子-κB(NF-κB)表达增强,水通道蛋白4(AQP4)表达增强,反映局部肺组织炎症及肺水肿等病理变化程度。④综合以上结果说明:生物性致病因子(流感病毒感染成功)是病证结合动物模型形成的关键,湿热状态下病毒感染小鼠存在CD_4~+/CD_8~+比例倒置、细胞/体液免疫失衡等免疫功能紊乱、IFN-γ、NF-κB等炎症因子过度表达、肺泡壁细胞水液代谢失调,并造成肺组织病理学改变。症状及病理变化与流感病毒感染关系最为密切,高脂饮食、间断湿热环境应激在一定程度上影响了发病时间、病程及病理变化,可能对证型的发展方向有一定影响。
治法研究方面:①本实验中,清热化湿透表法对小鼠流感病毒性肺炎湿热证模型肺指数有一定抑制作用,但是与辛凉解表法、益卫固表法相比无明显的差异;②从免疫平衡角度分析:清热化湿透表法能够提高模型小鼠外周血CD_4~+T%,降低CD_8~+ T%,调节细胞免疫,维持免疫平衡,调节Th1/Th2类细胞平衡向Th2方向回归,该作用较益卫固表法作用更明显,而辛凉解表法效果不明显。推测整体增强并调节细胞免疫功能可能是该法抗流感病毒作用机理之一。③从炎症及病理变化角度分析:清热化湿透表法能够明显降低小鼠病毒感染后肺泡表面细胞炎症启动因子NF-κB阳性表达,此与其它两种治法相比有一定差异,推测调整机体细胞/体液免疫、减少致炎因子表达可能是该法的作用机制之一。④此外,模型鼠肺组织AQP4表达异常,清热化湿透表法与辛凉解表法、益卫固表法相比对此干预略有不同,尚不能肯定该治法是通过调节AQP4功能发挥作用的。
综上,清热化湿透表法、益卫固表法、辛凉解表法对小鼠流感病毒性肺炎湿热证模型有多方面、多靶点的作用机制。其中清热化湿透表法在增强细胞免疫、调节Th1/Th2细胞平衡、降低炎症启动因子方面具有优势。对于复合湿热因素后造成的小鼠流感病毒性肺炎,三种治法均能有效降低肺指数,减轻病变程度。蒿芩清胆汤、玉屏风散能够更明显地增强机体T细胞免疫功能,调节Th1/Th2类细胞平衡向Th2方向回归。而银翘散则主要从抑制炎症、减轻肺水肿方面起作用。
结论
综上所述,岭南地区急性呼吸道感染者辨证以风温为主,证候多挟湿,辨病与辨证结合,因地制宜采用综合治法治疗呼吸道病毒感染具有一定优势。以病毒感染结合饮食、环境构建湿热证模型中,病毒感染引起机体免疫功能紊乱是模型成功的关键,湿热状态下病毒感染小鼠存在免疫功能紊乱、炎症因子过度表达、水液代谢失调及病理改变(湿热共同的靶点)。以蒿芩清胆汤为代表的清热化湿透表法能有效降低肺指数,减轻病变程度,与益卫固表法(玉屏风散)、辛凉解表法(银翘散)相比差异不明显。但与其它两种治法相比,该法在调节机体T细胞免疫,调整Th1/Th2细胞平衡,抑制炎症因子表达等环节具有一定优势。提示该法的调节免疫、抑制炎症启动因子的表达等可能是与益卫固表法(增强免疫)及辛凉解表法(抗病毒、抗炎)不完全相同的作用靶点。本研究为呼吸道病毒感染辨证从湿热论治提供了临床和实验室依据。
Objective
In recent years, herbal treatments show particular advantage in treating viral diseases. Especially to say, complex prescriptions have good effects in multi-direction and regulation of immunity, with the aim to getting rid of pathogen without disturbing the body's normal function. Respiratory viral infection is the most common diseases in South of china. Influenza and parainfluenza viruses are detected among adult cases. Researches of TCM have showed that respiratory viral infection in southern china is of heat and dampness character. The therapeutic principles are made according to the pathogen and syndrome, among which clearing heat and dampness, clearing heat and toxin, building up and tonifying immunity are very popular. Some researches showed that the effects of these principles are inhibiting virus cloning, regulating the immunity, or reducing inflammation etc. But there are many unsettled problems in this field.
Hao Qin Qing Dan decotion(HQQD) ,which represents clearing heat and dampness, relieving superficies therapeutic principle, is wildly used in treatments on respiratory viral diseases and Febrile diseases in Southern China. To clarify the connotation and epitaxy of heat-dampness syndrome in respiratory viral infection, and explore the mechanism of clearing heat and dampness, relieving superficies therapeutic principle, we use a influenza virus infected mouse model with composite factors in our experiments to imitate the syndrome of respiratory infection in Southern China. Comparing with the other two therapeutic principles, we want to study the mechanism of the clearing heat and dampness, relieving superficies therapeutic principle in this model and give more certifications to use this principle in treating respiratory viral diseases.
Method: According to the normal heat-dampness animal model, on the basis of fat animal feeds and heat-dampness conditions, mice are infected by influenza virus (H_1N_1) by dropping. Lung indexes are used in our research to evaluate the anti-virus effect of experimental decoctions. Different lymphocyte groups of model mice are detected by FCM. IFN- y and IL-4 level in the blood are tested by ELISA to estimate Thl/Th2 and immunity balance. NF-KB and aquaporin 4 are detected by immunohistochemical method in lung. We want to explore the main direction of the therapeutic principle by comparing it with Ying Qiao San decotion(HQS, represent relieving superficies by acrid-cold herbs) and Yu Ping Fen San decotion(YPFS, represent building up and tonifying immunity). In our clinical research, 101 adult cases of upper respiratory infection are colleted, syndrome and prescription character of these cases are analyzed and summarized.
Results and Analysis:
About clinical study: 101 adult cases of acute upper respiratory infection from June to August in 2007 are collected and analyzed. As a result: 1. onset of the disease is closely related to the solar term. Attack rate is highest during Grain in Ear. The main diagnosis is anemogenous febrile disease (wind-heat with dampness, syndrome type); 2.Fever, headache, limbers with aching pain, sore throat are most common symptoms, and the most common simultaneous phenomenon are anorexia, dry mouth, red tongue with thick float;3. Clearing heat and toxin, relieving superficies by acrid-cold herbs and opening the inhibited lung-qi to dissipate dampness are very popular in the prescriptions according to differentiation of symptoms and signs. Sang Ju Yin(SJY),Yin Qiao San(YQS),which represent the main principle in Febrile Disease, are wildly used in Southern China. Pharmaco-experiments have proved that isatis root, dyers woad leaf, forsythia suspense are effective in inhibiting viruses in vitro and vivo. These herbs be useful in antibiosis, relieving fever and analgesia, too. 4. The following local herbs are very popular in prescriptions, such as Operculate cleistocalyx(function: clear summer heat and dampness), snakegourd fruit(function: lower adverse-rising qi and disspate phlegm), roughhaired holly root(function:clear toxin and relieving sore throat ),musslinite etc.
About the animal model: 1. the symptoms and pathogenic changes are closely related to influenza virus infection. Fat feeds and intmt heat conditions, not act as the main factors, influence the onset and pathogenic changes of disease; 2. The the balance of T cell groups is broken after virus infection, percent of CD_4~+T increases and CD_4~+T decreases, the ratio of CD_4~+ and CD_8~+ inverts. Virus infection inhibits the immunity system of mice. Meanwhile, the level of IFN-γincreased and IL-4 keep still. Th1/Th2 balance is directed to Th1. These changes are same between model mice and virus-infected mice; 3. Expression of NF-κB and AQP4 in lung like enhanced in model mice, to show the inflammation and edema in local lung tissues. The slight difference between model mice and virus infected mice is of no significance; 4. To sum up, living causative agent (influenza virus infected successfully) is the key factor to set up the animal model. With heat and dampness factors, virus-infected mice's cell/humoral immunity system is interrupted, some inflammatory factors,such as IFN-γ、NF-κB etc, are over expressed and water metabolism of alveolar wall cells is irregular, which leads to pathogenic injury.
About the therapeutic principle: 1. HQQD(clear heat and dampness, relieve superficies therapeutic principle) is effective in inhibiting the pathogenic injury of model mice, which is similar with YQS(relieve superficies by acrid-cold herbs) and YPFS(build up and tonify the immunity); 2.HQQD can increase the percent of CD_4~+T and decrease the percent of CD_8~+T, advance cell immunity and keep the immunity balcance. This part is similar with YPFS which represent building up and tonify immunity therapeutic principle. And we presume that accelerating cell immunity maybe one of mechanism that anti-virus function of the principle; 3. HQQD can inhibit the increase of IFN-γand direct the Th1/Th2 to Th2, this function is more significant than YPFS. YQS is not significant in this aspect. We presume that regulating cell/humoral immunity, reducing inflammatory factors expression maybe one of mechanism that anti-virus function of the principle; 4. HQQD(clear heat and dampness to relieve superficies) reduce the positive expression of NF-κB obviously. And there is no difference between HQQD,YQS and YPFS. Furthermore, three decotions(three different therapeutic principle) can reduce the expression of AQP4 on the alveolar wall cells, but YQS and YPFS are more effective.
To sum up, three different therapeutic principles are effective in reducing lung index, reliving pathogenesis. HQQD and YPFS are more effective in strengthen T cell immunity; HQQD, which turns Th1/Th2 to Th2, is more effective than YPFS in regulate Th1/Th2 balance. YQS's function may exist in inhibiting and reducing inflammation and edema of lung. As a result, three therapeutic principles(HQQD,YQS,YPFS) have multiple function targets in treating influenza A viral pneumia. HQQD(clear heat and dampness, relieve superficies) has advantage in strengthening cell immunity, reducing inflammatory factors, and regulating Th1/Th2 balance.
Conclusion
On the whole, acute upper respiratory infection in Southern China belongs to anemogenous febrile disease, and the main syndrome type is wind-heat with dampness. Different therapeutic principle has its advantage in treating respiratory vrial infection according to differentiation of symptoms and signs in different area. As far as heat-dampness animal model in our research are concerned, immunity system derangement after virus infection is key pathogenesis in heat-dampness mouse influenza pneumia model comparing with fat feeds and hot condition. The state of model mouse(immunity derangement, over expression of inflammatory factors, disharmony of water metabolism ) may be the target of heat and dampness. HQQD(clearing heat and dampness, relieving superficies therapeutic principle) can effectively reduce the lung index, relieve the pathogenic changes. HQQD can regulate T cell immunity and Th1/Th2 balance significantly, comparing with YQS and YPFS. Our experimental result hints that HQQD probably not the same role with the other two principles. Our research provides more evidences to respiratory viral infection treatment by using heat and dampness theory.
近年来,中医药在治疗病毒性疾病方面逐步体现出独特的优势,特别是中药复方多靶点作用,整体调节机体免疫,达到“祛邪不伤正”的目的。呼吸道病毒感染是岭南地区临床常见病、多发病。病毒学检测发现,成人以流感病毒、副流感病毒感染为主。本病的临床治疗采用辨病与辨证相结合,临证治疗相当灵活。常用的中医治法有清热化湿、清热解毒、益卫固表等,其作用机理与抑制病毒复制、调节机体免疫、减少局部炎症渗出等有关。上述研究加深了对运用中医药治疗呼吸道病毒感染的认识,但还存在不少亟待解决的问题。本病的发生与病毒、机体、环境等多种因素相关,不同治法在疾病的不同阶段发挥作用,如何在治法研究中体现“辨证”的思路,有待进一步探讨。
研究表明,岭南地区呼吸道病毒感染从中医辨证具有易化热挟湿的特点。本研究前期的临床调查亦表明急性上呼吸道感染发病初起即易挟湿。清热化湿透表法是该地区常用外感热病治法之一,其中用于气分湿热证的蒿芩清胆汤除了具有清热化湿作用外,方中青蒿更能透邪达表,是清热化湿透表的代表方之一。以该方为主的经验治法广泛应用于岭南地区的呼吸道病毒感染治疗,具有一定地域特点,体现了中医辨证“三因制宜”的思路。为进一步明确岭南地区呼吸道病毒感染湿热证的内涵和外延,探讨清热化湿透表法的作用机理,我们进行了以下研究,以期为临床辨证运用清热化湿透表法治疗呼吸道病毒感染提供理论、临床和实验研究依据。
研究方法
本研究通过文献回顾及对101例急性上呼吸道感染患者的证候特点及用药特点的观察,在传统湿热证动物模型基础上,采用“肥甘饮食+湿热环境+流感病毒感染”方法,构建小鼠流感病毒性肺炎湿热证模型。以蒿芩清胆汤代表清热化湿透表法,以肺指数初步评价该法的治疗效果,以流式细胞术检测模型小鼠T淋巴细胞CD_3~+/CD_4~+,CD_8~+T亚群变化,以ELISA法检测血清IFN-γ及IL-4水平,通过Th1/Th2类细胞平衡变化评估机体免疫功能,采用免疫组织化学法检测肺组织炎症因子NF-κB及水通道蛋白AQP4的阳性表达,探讨清热化湿透表法干预病毒性肺炎湿热证的作用方向,并与银翘散(辛凉解表法)、玉屏风散(益卫固表法)比较异同。
研究结果及分析
模型构建方面:①造模成功后,模型小鼠体重减轻、肛温升高,蜷缩懒动、反应迟钝、呼吸急促甚则口唇紫绀,肺组织发生炎性实变。模型鼠症状的出现较单纯病毒感染小鼠滞后,似有减轻趋势。②模型小鼠感染病毒后,T淋巴细胞亚群比例失调,CD_4~+T%降低,CD_8~+T%升高,CD_4~+/CD_8~+比例倒置,病毒感染造成小鼠免疫系统抑制;致炎因子IFN-γ分泌显著增加,而IL-4分泌水平未见明显变化,Th1/Th2细胞因子平衡向Th1方向偏移,与病毒组相比差异不显著。③模型小鼠肺泡壁细胞核因子-κB(NF-κB)表达增强,水通道蛋白4(AQP4)表达增强,反映局部肺组织炎症及肺水肿等病理变化程度。④综合以上结果说明:生物性致病因子(流感病毒感染成功)是病证结合动物模型形成的关键,湿热状态下病毒感染小鼠存在CD_4~+/CD_8~+比例倒置、细胞/体液免疫失衡等免疫功能紊乱、IFN-γ、NF-κB等炎症因子过度表达、肺泡壁细胞水液代谢失调,并造成肺组织病理学改变。症状及病理变化与流感病毒感染关系最为密切,高脂饮食、间断湿热环境应激在一定程度上影响了发病时间、病程及病理变化,可能对证型的发展方向有一定影响。
治法研究方面:①本实验中,清热化湿透表法对小鼠流感病毒性肺炎湿热证模型肺指数有一定抑制作用,但是与辛凉解表法、益卫固表法相比无明显的差异;②从免疫平衡角度分析:清热化湿透表法能够提高模型小鼠外周血CD_4~+T%,降低CD_8~+ T%,调节细胞免疫,维持免疫平衡,调节Th1/Th2类细胞平衡向Th2方向回归,该作用较益卫固表法作用更明显,而辛凉解表法效果不明显。推测整体增强并调节细胞免疫功能可能是该法抗流感病毒作用机理之一。③从炎症及病理变化角度分析:清热化湿透表法能够明显降低小鼠病毒感染后肺泡表面细胞炎症启动因子NF-κB阳性表达,此与其它两种治法相比有一定差异,推测调整机体细胞/体液免疫、减少致炎因子表达可能是该法的作用机制之一。④此外,模型鼠肺组织AQP4表达异常,清热化湿透表法与辛凉解表法、益卫固表法相比对此干预略有不同,尚不能肯定该治法是通过调节AQP4功能发挥作用的。
综上,清热化湿透表法、益卫固表法、辛凉解表法对小鼠流感病毒性肺炎湿热证模型有多方面、多靶点的作用机制。其中清热化湿透表法在增强细胞免疫、调节Th1/Th2细胞平衡、降低炎症启动因子方面具有优势。对于复合湿热因素后造成的小鼠流感病毒性肺炎,三种治法均能有效降低肺指数,减轻病变程度。蒿芩清胆汤、玉屏风散能够更明显地增强机体T细胞免疫功能,调节Th1/Th2类细胞平衡向Th2方向回归。而银翘散则主要从抑制炎症、减轻肺水肿方面起作用。
结论
综上所述,岭南地区急性呼吸道感染者辨证以风温为主,证候多挟湿,辨病与辨证结合,因地制宜采用综合治法治疗呼吸道病毒感染具有一定优势。以病毒感染结合饮食、环境构建湿热证模型中,病毒感染引起机体免疫功能紊乱是模型成功的关键,湿热状态下病毒感染小鼠存在免疫功能紊乱、炎症因子过度表达、水液代谢失调及病理改变(湿热共同的靶点)。以蒿芩清胆汤为代表的清热化湿透表法能有效降低肺指数,减轻病变程度,与益卫固表法(玉屏风散)、辛凉解表法(银翘散)相比差异不明显。但与其它两种治法相比,该法在调节机体T细胞免疫,调整Th1/Th2细胞平衡,抑制炎症因子表达等环节具有一定优势。提示该法的调节免疫、抑制炎症启动因子的表达等可能是与益卫固表法(增强免疫)及辛凉解表法(抗病毒、抗炎)不完全相同的作用靶点。本研究为呼吸道病毒感染辨证从湿热论治提供了临床和实验室依据。
Objective
In recent years, herbal treatments show particular advantage in treating viral diseases. Especially to say, complex prescriptions have good effects in multi-direction and regulation of immunity, with the aim to getting rid of pathogen without disturbing the body's normal function. Respiratory viral infection is the most common diseases in South of china. Influenza and parainfluenza viruses are detected among adult cases. Researches of TCM have showed that respiratory viral infection in southern china is of heat and dampness character. The therapeutic principles are made according to the pathogen and syndrome, among which clearing heat and dampness, clearing heat and toxin, building up and tonifying immunity are very popular. Some researches showed that the effects of these principles are inhibiting virus cloning, regulating the immunity, or reducing inflammation etc. But there are many unsettled problems in this field.
Hao Qin Qing Dan decotion(HQQD) ,which represents clearing heat and dampness, relieving superficies therapeutic principle, is wildly used in treatments on respiratory viral diseases and Febrile diseases in Southern China. To clarify the connotation and epitaxy of heat-dampness syndrome in respiratory viral infection, and explore the mechanism of clearing heat and dampness, relieving superficies therapeutic principle, we use a influenza virus infected mouse model with composite factors in our experiments to imitate the syndrome of respiratory infection in Southern China. Comparing with the other two therapeutic principles, we want to study the mechanism of the clearing heat and dampness, relieving superficies therapeutic principle in this model and give more certifications to use this principle in treating respiratory viral diseases.
Method: According to the normal heat-dampness animal model, on the basis of fat animal feeds and heat-dampness conditions, mice are infected by influenza virus (H_1N_1) by dropping. Lung indexes are used in our research to evaluate the anti-virus effect of experimental decoctions. Different lymphocyte groups of model mice are detected by FCM. IFN- y and IL-4 level in the blood are tested by ELISA to estimate Thl/Th2 and immunity balance. NF-KB and aquaporin 4 are detected by immunohistochemical method in lung. We want to explore the main direction of the therapeutic principle by comparing it with Ying Qiao San decotion(HQS, represent relieving superficies by acrid-cold herbs) and Yu Ping Fen San decotion(YPFS, represent building up and tonifying immunity). In our clinical research, 101 adult cases of upper respiratory infection are colleted, syndrome and prescription character of these cases are analyzed and summarized.
Results and Analysis:
About clinical study: 101 adult cases of acute upper respiratory infection from June to August in 2007 are collected and analyzed. As a result: 1. onset of the disease is closely related to the solar term. Attack rate is highest during Grain in Ear. The main diagnosis is anemogenous febrile disease (wind-heat with dampness, syndrome type); 2.Fever, headache, limbers with aching pain, sore throat are most common symptoms, and the most common simultaneous phenomenon are anorexia, dry mouth, red tongue with thick float;3. Clearing heat and toxin, relieving superficies by acrid-cold herbs and opening the inhibited lung-qi to dissipate dampness are very popular in the prescriptions according to differentiation of symptoms and signs. Sang Ju Yin(SJY),Yin Qiao San(YQS),which represent the main principle in Febrile Disease, are wildly used in Southern China. Pharmaco-experiments have proved that isatis root, dyers woad leaf, forsythia suspense are effective in inhibiting viruses in vitro and vivo. These herbs be useful in antibiosis, relieving fever and analgesia, too. 4. The following local herbs are very popular in prescriptions, such as Operculate cleistocalyx(function: clear summer heat and dampness), snakegourd fruit(function: lower adverse-rising qi and disspate phlegm), roughhaired holly root(function:clear toxin and relieving sore throat ),musslinite etc.
About the animal model: 1. the symptoms and pathogenic changes are closely related to influenza virus infection. Fat feeds and intmt heat conditions, not act as the main factors, influence the onset and pathogenic changes of disease; 2. The the balance of T cell groups is broken after virus infection, percent of CD_4~+T increases and CD_4~+T decreases, the ratio of CD_4~+ and CD_8~+ inverts. Virus infection inhibits the immunity system of mice. Meanwhile, the level of IFN-γincreased and IL-4 keep still. Th1/Th2 balance is directed to Th1. These changes are same between model mice and virus-infected mice; 3. Expression of NF-κB and AQP4 in lung like enhanced in model mice, to show the inflammation and edema in local lung tissues. The slight difference between model mice and virus infected mice is of no significance; 4. To sum up, living causative agent (influenza virus infected successfully) is the key factor to set up the animal model. With heat and dampness factors, virus-infected mice's cell/humoral immunity system is interrupted, some inflammatory factors,such as IFN-γ、NF-κB etc, are over expressed and water metabolism of alveolar wall cells is irregular, which leads to pathogenic injury.
About the therapeutic principle: 1. HQQD(clear heat and dampness, relieve superficies therapeutic principle) is effective in inhibiting the pathogenic injury of model mice, which is similar with YQS(relieve superficies by acrid-cold herbs) and YPFS(build up and tonify the immunity); 2.HQQD can increase the percent of CD_4~+T and decrease the percent of CD_8~+T, advance cell immunity and keep the immunity balcance. This part is similar with YPFS which represent building up and tonify immunity therapeutic principle. And we presume that accelerating cell immunity maybe one of mechanism that anti-virus function of the principle; 3. HQQD can inhibit the increase of IFN-γand direct the Th1/Th2 to Th2, this function is more significant than YPFS. YQS is not significant in this aspect. We presume that regulating cell/humoral immunity, reducing inflammatory factors expression maybe one of mechanism that anti-virus function of the principle; 4. HQQD(clear heat and dampness to relieve superficies) reduce the positive expression of NF-κB obviously. And there is no difference between HQQD,YQS and YPFS. Furthermore, three decotions(three different therapeutic principle) can reduce the expression of AQP4 on the alveolar wall cells, but YQS and YPFS are more effective.
To sum up, three different therapeutic principles are effective in reducing lung index, reliving pathogenesis. HQQD and YPFS are more effective in strengthen T cell immunity; HQQD, which turns Th1/Th2 to Th2, is more effective than YPFS in regulate Th1/Th2 balance. YQS's function may exist in inhibiting and reducing inflammation and edema of lung. As a result, three therapeutic principles(HQQD,YQS,YPFS) have multiple function targets in treating influenza A viral pneumia. HQQD(clear heat and dampness, relieve superficies) has advantage in strengthening cell immunity, reducing inflammatory factors, and regulating Th1/Th2 balance.
Conclusion
On the whole, acute upper respiratory infection in Southern China belongs to anemogenous febrile disease, and the main syndrome type is wind-heat with dampness. Different therapeutic principle has its advantage in treating respiratory vrial infection according to differentiation of symptoms and signs in different area. As far as heat-dampness animal model in our research are concerned, immunity system derangement after virus infection is key pathogenesis in heat-dampness mouse influenza pneumia model comparing with fat feeds and hot condition. The state of model mouse(immunity derangement, over expression of inflammatory factors, disharmony of water metabolism ) may be the target of heat and dampness. HQQD(clearing heat and dampness, relieving superficies therapeutic principle) can effectively reduce the lung index, relieve the pathogenic changes. HQQD can regulate T cell immunity and Th1/Th2 balance significantly, comparing with YQS and YPFS. Our experimental result hints that HQQD probably not the same role with the other two principles. Our research provides more evidences to respiratory viral infection treatment by using heat and dampness theory.
引文
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