柴石退热颗粒对乙型脑炎病毒体外感染BHK细胞的影响及流行性乙型脑炎辨证规律的研究
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摘要
目的
     以中医基础理论为指导,遵照循证医学原则,通过大样本、多中心、随机对照的临床研究,梳理流行性乙型脑炎不同证型的中医症候要素及结局转归,总结其病机、传变及预后规律,验证中医预案的科学性和准确度,对2011年乙脑中医预案进行完善。运用不同中医方案治疗乙脑患者,对临床疗效进行分析,总结乙脑中医治法,筛选有效中成药。从而系统深化乙脑的中医药理论和临床认识,为中医药治疗病毒性传染病规范化体系的建立提供科技支撑。
     方法
     在2012年6月至2012年10月间,现场考察确诊为流行性乙型脑炎患者284例进行前瞻性临床研究。参照2009年中医药行业专项“中医药防治流行性乙型脑炎临床规律与诊疗方案的研究”(简称乙脑项目)课题组制定的中医预案,在急性期将所有病例分为轻型(毒蕴肺胃证)、普通型(毒损脑络证)、重型(毒陷心包证)、极重型(阴阳衰竭证),填写《流行性乙型脑炎病例调查表》(CRF表),记录人口学资料,各症状、体征发生的程度、频次、出现时间、消失(或明显减轻)时间,中医证候及舌脉信息,中医辨证分型,住院时间,用药记录,结局判断,随访记录等。将CRF表记录的中医症候要素按照不同证型进行整理和排序。
     根据“乙脑项目”临床实施方案的分组方法,轻型、普通型病人纳入随机组,重型、极重型纳入队列组。通过计算机软件(中国中医科学院临床评价中心中央随机系统)生成随机数字,具体采用区组与动态随机结合方法,应用SAS9.1.3统计软件PROC PLAN程序进行随机化分配,将受试者所接受处理随机安排,分为治疗组和对照组。
     对照组用西医常规综合治疗;治疗组在对照组治疗的基础上加用中医药:①中医辨证处方②柴石退热颗粒③抗病毒口服药④喜炎平注射液。比较两种治疗方法的临床疗效、对高热、神昏、抽搐(乙脑三大主症)的治疗效果、对预后的影响。尤其以发热为切入点,观察中医治疗方案中有效药物的临床疗效。
     定量资料的描述将计算均数、标准差、中位数、最小值、最大值。定性资料描述各类的例数及百分数。
     统计学处理由SPSS16.0软件完成,两组可比性分析,定性资料采用卡方检验/Fisher精确概率法/Wilcoxon秩和检验。定量资料符合正态分布用t检验(方差不齐时进行t’检验),不符合正态分布用Wilcoxon秩和检验,多组间差异采用单因素方差分析(one-way ANOVA),两两组间比较用LSD法。P<0.05为所检验的差别有统计意义,P<0.01指差别显著。
     结果
     1.病例资料:2012年本研究共观察乙脑病例284例,在急性期进行中医辨证分型:毒蕴肺胃型61例,占总例数的21.48%;毒损脑络型114例,占总例数的40.14%;毒陷心包型98例,占总例数的34.51%;阴阳衰竭型11例,占总例数的3.87%。年龄最小为3月,最大为15岁,平均年龄5岁7月;男女比例为1.63:1。
     2.传变及预后规律:毒蘊肺胃证病在卫表,易于治愈;毒损脑络证为气分热甚,邪在中焦,或气营同病、有生风之兆,大部分临床可治愈。毒陷心包证为热毒内陷营分,耗气伤津,兼或营血同病,动风耗血,或兼湿热并重、痰湿内生,约30%病患预后欠佳;阴阳衰竭见热伤气阴,易成内闭外脱、亡阴亡阳之象,病情危重,须积极抢救生命。并且,乙脑病情变化较一般温病更迅速更危重。
     3.恢复期,近40%病情较轻者中医症候要素:纳差、便秘、乏力少言、低热、多汗、口渴、胸闷、恶心,或伴肢体痉挛、颈强;舌象:舌暗红、少苔、质干,舌淡暗、苔白厚、舌体胖大;脉象:脉细无力。
     另有23.8%例病情较重者中医症候要素:发热易反复、伴抽搐、神志不清、头痛,肢体强直、神志呆滞、记忆力下降、肢体瘫痪、失语、面肌瘫痪、性格改变;舌象:舌淡或暗红、苔白厚,舌暗或紫、有瘀斑瘀点;脉象:细涩、弦细。
     4.将两期五种中医辨证分型症候要素与2011年中医预案分型要素对比,发现两点不同之处:①部分发热者伴胸闷、纳差、便溏、苔薄白或白厚,提示暑热挟湿致病。②部分邪犯气营或营血同病者,病程较长、病情偏重,在恢复期有18%的病例出现肢体强直、神志呆滞、肢体瘫痪、失语、面肌瘫痪、记忆力下降等症状,观察结束时仍有11.6%的病例出现残障,表现出痰瘀阻滞,经脉失养之证。
     5.临床疗效:观察结束时,分析中西医结合组与对照西医组的疗效,结果显示,随机组疗效:治疗组治愈率90.6%,好转率7.69%,无效率1.71%;对照组治愈率74.14%,好转率18.97%,无效率6.7%。队列组疗效:治疗组治愈率67.53%,好转率23.37%,无效率9.09%;对照组治愈率46.88%,好转率40.62%,无效率12.5%。随机组和队列组分别组内比较,治疗组疗效明显优于对照组,差异皆有统计学意义(P<0.05)。
     6.三大主症疗效分析:
     随机组,治疗组退热中位数为72小时,对照组为96小时;队列组,治疗组退热中位数为96小时,对照组为144小时;通过生存分析法比较,两研究组加用中医治疗组退热趋势快于对照组,差别有统计学意义(P<0.05)。
     随机组,治疗组神志完全复苏时间中位数为72小时,对照组为120小时;队列组,治疗组退热中位数为85小时,对照组为132小时;通过生存分析法比较,两研究组加用中医的治疗组神志恢复正常趋势快于对照组,差别有统计学意义(P<0.05)。
     筛选CRF表抽搐各类型的止痉情况,逐天进行记录,并与基线对比(观察第一天),结果显示:随机组治疗第三天,两组较基线均有缓解,且治疗组缓解优于对照组;队列组治疗第五天,两组较基线均有缓解,且治疗组缓解优于对照组;差别有统计学意义(P<0.05)。
     7.284例乙脑患者,治疗组194例均按照中医治疗方案执行,中药使用率达100%,治愈率为83.6%,其中服用柴石退热颗粒有44例,治愈率为93.18%;中医辨证处方33例,治愈率为81.82%;抗病毒口服液30例,治愈率73.33%、喜炎平注射液120例,治愈率84.16。
     8.其他中成药使用程度依次是:炎琥宁注射液、热毒宁注射液、痰热清注射液、醒脑静注射液、安宫牛黄丸、四磨汤、保和丸等。中医辨证处方主方使用程度依次是:白虎汤、银翘散、三仁汤、清营汤、泻白散、竹叶石膏汤、沙参麦冬汤、羚角钩藤汤等。
     9.柴石退热颗粒治疗乙脑44例,平均退热起效时间为10.23±6.24h,平均完全退热时间为68.92±18.54h;对照组治疗乙脑31例,平均退热起效时间为12.53±7.31h,平均完全退热时间为106.55±24.83h;两者分别经统计学分析,差别有统计学意义(P<0.05)。
     10.柴石退热颗粒治疗乙脑,对队列组病人治愈率优势大于随机组,在退热的同时,可以有效缓解神昏、抽搐、缩短疗程、降低后遗症发生率。与对照组比较,差别有统计学意义(P<0.05)。
     11.安全性检测:两组治疗过程中均无严重的肝肾功能损害。不良事件发生与药物无关,极少数病例出现胃肠道反应,对症处理后消失。
     结论:
     本研究表明,流行性乙型脑炎流行于夏季,多见于学龄前及学龄儿童,男性略多于女性。乙脑中医病机及传变规律基本符合卫气营血传变特点,乙脑传变更迅速,病情较一般温病更危重。通过对2012年临床中医症候要素的梳理,并与乙脑中医预案内容进行对比,证明乙脑项目中医预案对乙脑的分期分型辨证治疗符合临床实际,具有较高的科学性和准确性。同时发现2012年乙脑致病的特点为暑热挟湿,治疗原则在“清热解毒”基础上,注意湿热并除,加用淡渗利湿中药。并对2011年中医预案恢复期加以优化,增加肝肾阴虚,痰瘀阻络证。
     我们用大量的临床数据和现代科研技术方法证实中医药在防治乙脑这种病毒感染类传染病中作用突出,在缓解临床症状、提高治愈率、减少后遗症等方面具有明显优势。筛选出有效中成药如喜炎平注射液、醒脑静注射液、柴石退热颗粒、抗病毒口服液、安宫牛黄丸等等。其中,柴石退热颗粒临床治疗乙脑效果主要表现在退热起效时间快、退热效力强、有效控制抽搐及减轻意识障碍,缩短疗程,同时,可以降低重症乙脑后遗症的发生。对重症病例的治疗优势尤为明显。
     由此可见,中医药治疗再发的经典传染病有着非常明显的优势。充分发挥中医药特色,深入发掘先辈治疗经验,对防治此类传染病有非常重要的意义。
     目的
     以体外细胞培养技术为支撑,在细胞水平上对柴石退热颗粒抗乙脑病毒(JEV)的作用功效及作用靶点进行初步探索,为中药抗乙脑病毒提供客观实验依据,从而扩大柴石退热颗粒抗病毒谱,同时,为下一步进行分子基因水平的研究以及整体动物试验提供科学依据。
     方法
     利用BHK-21传代细胞株建立乙脑病毒体外细胞感染模型,检测实验用病毒的活性,以BHK细胞检测药物细胞毒性,再以不同浓度的药物干预细胞,通过不同给药方式,使用同步感染法和空斑减少计数测定病毒滴度,分别考察药物对JEV感染细胞的抑制作用,对病毒的直接杀伤作用,以及药物对JEV吸附、侵入细胞的阻断作用和细胞内增殖的抑制作用。另设立利巴韦林药物对照组和病毒感染对照组。
     所得计量数据以均数+标准差(x±s)表示,多组间差异采用单因素方差分析(one-way ANOVA),两两组间比较用LSD法;计数资料用X2检验,所有统计采用SPSS16.0统计软件计算,P<0.05为有统计学意义,P<0.01有显著意义。
     结果
     1.乙脑感染细胞体外模型的建立:本实验所用JEV为强毒株,经活化后接种于BHK-21细胞,使细胞在48-72小时间发生细胞膜变圆、细胞染色质加深、细胞脱落、凋亡等病理变化,并且病毒在细胞传代3次后病变时间趋向稳定。
     2.药物细胞毒性实验:利巴韦林随着药物浓度的递增,显示出抑制细胞扩增的效应,在1600ug/ml浓度下可将细胞生长速度减慢三倍,但不引起细胞凋亡;而柴石退热颗粒对细胞的毒性表现在对细胞的杀伤,当药液浓度达到320ug/ml时,大部分BHK-21细胞表现出坏死。柴石退热颗粒药液的最大无毒浓度(TD0)为160ug/ml,利巴韦林10ug/ml即可达到对乙脑病毒的抑制作用。
     3.药物对病毒复制的抑制作用:柴石退热颗粒40,80,160ug/ml组、利巴韦林组与病毒对照组相比,均有显著性差异(P<0.01);柴石退热颗粒160ug/ml组与柴石退热颗粒40ug/ml组相比,有统计学差异(P<0.05);中药160ug/ml组与利巴韦林组相比,差别无统计学意义。
     4.柴石退热颗粒对JEV的直接杀伤作用:柴石退热颗粒160ug/ml组较病毒对照组,病毒滴度下降了约53333.3PFU/ml,但经统计学分析,差别无意义。
     5.柴石退热颗粒抑制JEV复制的作用靶点研究:柴石退热颗粒160ug/ml组预处理细胞后感染病毒能降低其复制率,与病毒对照组,有统计学差异(P<0.05);柴石退热颗粒80、160ug/ml组在病毒侵染细胞时给药,能一定程度阻断JEV的入侵,较病毒对照组,有统计学差异(P<0.05),中药两浓度之间对病毒入侵的影响无差别;柴石退热颗粒160ug/ml组作用于病毒在细胞内复制环节,对病毒复制的影响较病毒对照组,有统计学差异(P<0.05)。
     结论:
     本实验成功利用BHK-21传代细胞建立了乙脑病毒感染细胞的体外模型,观察柴石退热颗粒对乙脑病毒感染的抵制作用。结果显示,柴石退热颗粒在体外细胞模型上能较好的抑制乙脑病毒感染,且抑制作用与中药浓度呈正相关,高浓度组作用效果与利巴韦林相当。该药物抗JEV的作用机制表现在药物对JEV吸附、侵入细胞的阻断作用和细胞内增殖的抑制作用。同时,说明柴石退热颗粒对宿主细胞具有预防性保护作用。
Part1.
     Objectives:
     Based on traditional Chinese medicine theory, in accordance with theprincipleofevidence-based medicine, the clinical studyoflarge sample, multi center, randomized, TCM syndrome elements and prognosis out come of different syndrome type carding on J-E, summarizesits pathogenesis, transmission and prognosis ofTCM, verification plan scientifically and accurately, to improve the Chinese plan (2011). Application of different TCMtreatmentof JE patients, the clinical efficacy was analyzed, summarizedthe treatmentofTCMandChinesemedicine screening effective JE. Thetheory of TCM and clinicalunderstanding and deepen JE, to providescientific and technological support to establish the standard of Chinese medicine for the treatment of virus infectious diseasesystem.
     Methods:
     From June to October in2012, to inspect the diagnosed284cases of patientswithepidemicencephalitisB were prospectiveclinicalstudy. On2009Chinesemedicine industryspecial"Chinese med-icine in prevention and treatment of epidemic encephalitis Clin ical Regularity and treatment ofreference"(JE projects) Chines-e plan research group established in the acuteperiod, allcasesweredivided intomild (DUYun lung stomach), ordinarytype (toxinhurtingbraincollaterals), heavy (virus syndromeofinvasionofpericardium), very heavy (yinandYang failuresyndrome), fillinthe "JE cases questionnaire"(CRF), recorded demographicdata, symptoms, signsthe severity, frequency, time, disappear (or reduce) thetime, TCMsyndrome and thetongueandpulse information, TCMsyndromedifferentiation, hospitalization, medicalrecords, ending judgment, follow-uprecord. TCM syndrome elements of the CRF tablerecords were co-llected and sortedaccording to different syndromes.Accordingto the "method of grouping implementation JE project" clinicalprograms, light, ordinarytypepatients randomized group, heavy, ve-ry heavy inthe cohort. Through the computers of tware(producedby the center for clinical evaluation China Academy of traditional Chinese Medicine) to generate random numbers, the central randomization system, block and dynamic random combination metho-d, using the SAS9.1.3statistical software PROC PLAN program were random lyassigned, the subjects receiveed treatment with mach-ine arrangement,divided into treatment group and control group.
     Western medicine control group with routine western medical treatment. Traditional Chinese medicine in the treatment groupwere treated with traditional Chinese medicine:①TCM prescription,②Chai Shi antipyretic granule,③Oral anti-viral④Xiyangping Injection.To alleviate the influence on prognosis of efficacy, clinical efficacy, comparedoftwo treatment methods of fever, coma, convulsions. In particular, with fever as the breakthroughpoint, to observe the clinical curative effect of TCM therapy eff ective drugs.
     Quantitative data description will calculate the mean, standard deviation, median, minimumvalue,maximum value. Qualitative data to describe the number and percentage of cases.Statisticalanalysis by SPSS16.0software, two groups of comparability anal-ysis, qualitative data using chi square test/Fisher exact probability/Wilcoxon rank sum test. Quantitative data with normal distribution by the t test (t 'variance incomplete inspection), does not meet the normaldistribution using the Wilcoxon rank sumtest, differences among groups by single factor analysis of variance (one-way, ANOVA) between the two two groups with LSD method. P<0.05is considered to be statistically significant difference between the test, P<0.01means significant difference.
     Results:
     1.Case information:In2012,the study observed a total of284JE cases, syndrome differentiation in acute period: Du Yun lungand stomach type in61cases, accounting for21.48%of the totalcases; toxin damaging brain collaterals type in114cases,40.14%of the total number of cases; virus invasion of pericardium type in98cases,34.51%of the total number of cases; Yin and Yang failure type in11cases, accounting for3.87%of the total cases. The minimum age for March, a maximum of15years old, the average age is5years old in July; the male to female ratio was1.63:1.
     2.The transfer and prognosis of lung and stomach syndrome: poison accumulate in the Wei, and Wei Qi is not smooth, easy to cu re; toxin damaging brain collaterals is very hot gas, evil in middle Jiao, awind of trillion, most clinical cure.
     3.Toxic syndromeofinvasionofpericardium for heatcellulitesYingfen, consuming Qi and body fluidand blood disease, or camp,air consumption of blood, or both, and damp phlegm endogenous,abo-ut30%patients with poor prognosis; Yin and Yang and the failu-re to see to fry the Qi and Yin,into the inner closed outer de, Yin Yang Exhaustion like, in critical condition, to save lives.And, JEcondition change more quickly and more critical than thegeneral epidemic febrile disease.
     4.Both of the five kinds of TCM syndrome type of TCM syndrome elements and2011plan type elements comparison, found thattwo different parts:①fever with tightness in the chest, poor appetite, loose stools, moss thin white or whitethick, suggestingthat the heat dampness pathogen.The part of evil gas camp or the campblood disease, longer duration of illness, stress, during the recovery period of18%cases occurred in various neurological impairment in cash, were observed at the end there are still11.6%cases of disability, showing the phlegm, meridian dystrophysyndrome.
     5.the mass effect:The end of the observation, analysis of Chinese and Western medicine groupandthecontrolgroup ofWesternmedicine treatment, results show, random groups: the treatment group cure rate was90.6%, improvement rate was7.69%, effective rate1.71%; control group, the cure rate was74.14%, improvement rate was18.97%, effective rate6.7%. Effect of queue groups: treatment group, the cure rate was68.49%, improvement rate was23.29%, effective rate8.22%; control group, the cure rate was46.8 8%, improvement rate was40.62%, effecttive rate1.25%. Study group two, significantly better than the control group curative effect in the treatment group, there are significant differences(P<0.05).
     6.Clinical analysis of three main symotoms:Random group, pyretolysis median3days,4days in control group; cohort, pyretolysis median4days,6days in control group; through comparisonof survival analysis, two research group combined with TCM trea-tment group antifebrile trend faster than the control group, the difference was significant (P<0.05).
     Random group, the treatment group consciousness recovery time was72hours,120hours in the control group; cohort, pyretolysis a median of85hours,132hours in the control group; through comparison of production analysis, two research group combinedwith TCM treatment groupconsciousness recovery normal trend faster than the control group, the difference was significant(P<0.05).Screening of remission CRF tic each type method, were recorded, and compared with baseline(watchthefirst day), resultsshowed: treatmentof third random day group, two groups wererelieved, and the treatment group was better than that of the control group relief; cohort for fifth days, the two group compared with the base line was relieved, and the treatment remission groupthan in the control group; the difference was significant (P<0.05).
     7.284patients with epidemic encephalitis B patients,194cases in the treatment group were treated with traditional Chinese medicine traditionnal Chinese medicine according to the execution of the project, the use rate of100%, the total cure rate was83.6%,
     8.Other tranditonal Chinese msdicine: Potassium Sodium Deh-ydroandroandrographolide Succinate Injection, Reduning injection, Tanreqing injection, Xingnaojing Injection
     9.Chai Shi antipyretic granule in the treatment of epidemicencephalitis B in44cases, the mean feveronset time was10.23±6.24h, average completely antifebrile time was68.92±18.54h; the control group treatment for JE in31cases, the mean fever onset time was12.53±7.31h, average completely antifebrile time was106.55±24.83h respectively; the statistical analysis, the difference was significant (P<0.05).
     10. Chai Shi antipyretic granule in treatment of JE, the queue group cure rate of the patients more advantages than randomgroup, in the fever at the same time, can effectively alleviatethe coma,convulsions, shorten the course of treatment, reducethe incidence of sequelae. Compared with the control group, thedifference was significant (P<0.05).
     11. Safety inspection:no damage to liver and kidney functi-on, serious in two groups. Adverse eventsand drug free, veryfew cases of gastrointestinal reaction, disappeared after symptomatic treatment.
     Conclusion:
     This study shows that,epidemic encephalitis B in the sum-er, more common in preschool and school aged children. Je TCM pathogenesis and transmission rules accord with weiqiyingxuetransmission characteristiccs, even JE transmission changesrapidly, theillness is generally more severe febrile disease. By2012theclinical TCM syndrome elements analysis, and compared with the tr-aditional Chinese medicine syndrome differentiation of JE plan contents, prove JE projectChinese plans for the JE differential treatment accorded with clinical practice, it is of higher scientificity and accuracy. At the same time, found the2012JE pathogenic for summerheatdampness, treatment principlein the "detoxification" basis, pay attention to heat and removing, andlight penetration dampness herbs. And in2011the Chinese plan to optimize the recovery period,increase the liver kidney yin deficiency, phlegm and blood stasis blocking collaterals.
     We demonstrate that the Chinese medicine infection prominent role in prevention and treatment ofinfectious disease epidemicencephalitis B virus with a large number of clinical data and modernscientific and technical methods, in relieving clinical symptoms, improve the cure rate hasobviousadvantages, reduce sequela etc.. Screeningofeffective traditional Chinese medicine such asXiyanping injection, Xingnaojing injection, Chai Shi antipyretic granule, Antiviral Oral-Liquid, Angong Niuhuang Pill and so on. Among them, Chai Shi antipyretic granule clinical treatment effect is mainly manifested in the JE fever fast onset time, fever has strong effect, effective control of seizures andreduce the disturbance of consciousness, shorten the course of treatment, at the same time, can reduce severe encephalitis sequela occurred. Advantages of treating severe cases obviously.
     Objectives:
     In vitro cell culture technology, anti Japanese encephalitis virus on Chai Shi antipyretic granule at the cellular level (JEV) explores the role function and target, for traditional Chinese medicine toprovide objective experimental basis for the Anti Japanese encephalitis virus, thereby expandingthe Chai Shi antipyreticgranule antiviral spectrum, at the same time, to provide ascientific basis for the next step of molecular studies on the gene level and whole animal test.
     Methods:
     Using the established model of Japanese encephalitis virus infection of cells in BHK-21cell line, withthe activity of the virus detection experiments, using BHK cells to detect drug toxicity, drug intervention cells to differentconcentrations, by different administration, the use of synchronous infection assay and plaque reduction was determined by counting the number of virus titer,respectively study drugs on inhibition of JEV infection cells, direct killing effect on the virus, anddrugs on JEV adsorption, invasive cell inhibitory effect of blocking proliferation and intracellular.The other set of ribavirin drug control group and virus control group.
     The measurement data are mean+standard deviation,, differences among groups by singlefactor analysis of variance (one-way, ANOVA) between the two two groups with LSD method; count datausing X2test statistics, all statistics calculated by using statistical software SPSS16.0, P<0.05was statistically significant, significant P<0.01.
     Results:
     1. Estabilshment of cell model in vitro JEV infection: thisexperiment uses JEV is virulent, after the activation of inoculated in BHK-21cells, the cell membrane in the small48-72becameround, cell chromatin deepen, cell shedding, apoptosis and pathological changes, the virus in the cell lesiontime tends to be stable after3times.
     2.Drug toxicity experiment:ribavirin with increasing drug concentration, showing the effect of inhibition of cellexpansion, at concentration of1600ug/ml can be three times the cell growth slowed, but did not induce apoptosis; and Chai Shi antipyretic Granule on the cell toxicity in the killing of cells, when theconcentration reaches320ug/ml, the majority of BHK-21cells showed necrosis. Maximum non-toxicconcentration Chai Shi antipyreticgranule liquid (TD0) for160ug/ml, ribavirin10ug/ml can achievethe inhibition of Japanese encephalitis virus.
     3. The suppression of viral replication:Chai Shi comparedto40,80160group ug/ml, Leigh Bhave Lin antipyretic granule group and virus control group, there were significant differences (P<0.01); compared with Chai Shi antipyretic granule160ug/ml group and Chai Shi antipyretic granule40ug/ml group, there was statistically significant differentce (P<0.05); compared withthe high concentration of traditional Chinese medicine group andthe Leigh Bhave Lin group, the difference was not statisticallysignificant.
     4.Direct killing effect of Chai Shi on JEV: Chai Shi Chai Shi antipyretic granule160ug/mlgroup compared with the virus control group, virus titer decreased by about53333.3PFU/ml, but n osignificant difference by statistical analysis.
     5.Study on the effect of target inhibition of JEV replication Chai Shi antipyretic granule: To reduce the rate of virus replication pretreatmentofcells160ug/ml after ChaiShi antipyretic granule group, controlgroupand virus, therewasstatisticallysignificantdifference (P<0.05); ChaiShi antipyretic granule80160ug/ml group was administered in virus infected cells, to a certain extent blocked the JEV intrusion, compared with virus control group, there was statistically significant difference (P<0.05) effect on the virus, there is no difference between the twoconcentrations; Chai Shi antipyretic granule group160ug/ml replication link within the cell to the virus, its effect on viral replication is the virus control group, there was statistically significant difference (P<0.05).
     Conclusion:
     In this experiment,the successful use of BHK-21cell wasestablished in vitro cell model of JE virus infection, to observe the effect of antipyretic Granule on Chai Shi boycott of Japanese encephalitis virus infection. The results showed, Chai Shi antipyretic granule in vitro cell model can better inhibition of Japanese encephalitis virus infection, and the inhibition associated with traditional Chinese medicine concentration, high concentration group effect and ribavirin. The mechanisms of drug resistance performance of JEV cell entry blocking inhibition proliferation andintracellular drug on JEV adsorption. At the same time, the Chai Shi antipyretic granule hasprotective effects on the hostcell.
引文
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