Heavy Floor Grams Are Dropping Antiviral Experiment Research

Influenza is a common and frequent ly-ocurring disease which had been globally pandemic for many times and threatens human being’s health.A lot of accomplishments of the development of antiviral medicine and influenza vaccine have been achieved in modern medicine. However,due to the variation of the antigens of the influenza virus and the limitations of the medicine itself, there are not any specific ideal anti-influenza medicine on the clinic. Influenza belongs to the category of prevalent cold in traditional chinese medicine(TCM).Many prescriptions establised by TCM proved to be effective on the prevention and treatment of influenza. In this thesis, the medicine used in the experiment is CLKGD pill. It is a new compound preparation developed by shangdong university of traditional Chinese medicine, composed of two Chinese herbs:Paris polyphylla and Hypericum perforatum, which has the function of clearing heat and detoxication by reliving superficies to treat and prevent disease caused by respirotary tract infection like influenza. This preperation has showed good anti-influenza effects in clinical application.This thesis will further investigate the antiviral effects of CLKG pill.Objectives1To observe the inhibitory action and evaluate the antivivral effects of CLKGD pill against influenza A virus H1N1、 influenza A virus H3N2、 influenza RP8srain、 parainfluenza、 rhinovirus type14respiratory syncytial virus adenovirus type3and Coxsackievirus B3。2To investigate the protective effects of the pill on mice infected with influenza virus FM1from death and evaluate the effectiveness of it to treat "prevalent cold"caused by the invation of heat evils and epidemic toxins in TCM theory. 3To provide experimental basis for futher clinical application of CLKGD pills through acute toxicity test.Methods1Methods to study the antiviral effects of CLKG pills in vitro:Establish cell models infected by influenza A virus H1N1、influenza A virus H3N2、 influenza RP8srain、 parainfluenza、 rhinovirus type14、 respiratory syncytial virus、adenovirus type3and Coxsackievirus B3respectively. Four dosage groups of CLKG pills and cell control group (blank control group)、 virus control group、 ribavirin positive control group are established to investigate the effects of CLKG pill extracts on the viability of host cell of each virus by cytopathogenic effect (CPE).Detect the inhibitory effet of CLKG pill extracts to the neuraminidase of influenza A virus H1N1、 influenza A virus H3N2and influenza RP8srain by Neuraminidase Assay Kit2Methods to study the therapeutic effects of CLKG pillsKunming mice are divided into6groups randomly:normal control group、 virus control group、 CLKG groups (high Medium and low dose group)、 positive control group.Mice model of influenza was establised by dripping the virus fluid to the nose。The morbidity and mortality were observed after CLKG pill extracts were given to the mice model and the indexes of lung were measured.3Methods of acute toxicity test of CLKG pillsKunming mice were selected and divided into treatment group and saline solution group randomly。 The largest dose of drugs the mice can endure was given to the treatment group by intragastric administration every2hours,3times per day. After14weeks, symptoms, body signs and the weight of mice in each group were recorded and the LD50of the pill were measured.Results 1Results of antiviral effects in vitroCLKG pill extracts have certain inhibitory effects to influenza A virus H1N1、 influenza A virus H3N2、 influenza RP8srain、 parainfluenza rhinovirus type14respiratory syncytial virus adenovirus type3and Coxsackievirus B3and treatment indicator of CLKG pill extracts to those viruses are as follows:14.46,10.32,12.14,10.7,13.3,71.4,7.03,42.8.2Protective effects to influenza mice from death and lung indexesCompared to the control group, no mice was dead in the CLKG pill9.375mg/kg treatment group, the protection rate of whic is100%(P <0.05), while the difference was significant when compared to ribavirin group (P<0.05)。 The lung indexes of every CLKG treatment groups were lower than the model group (P<0.05), among which the CLKG18.75mg/kg treatment group was the lowest and the difference was significant compared to ribavirin group (P<0.05).3Results of acute toxicity test of CLKG pillsLD50of the pill could not be measured because no death of mice had happened. The largest dai ly dose given to the mice was450mg/kg equivalent to200times of adult human daily dose. After given the drugs, the intake of food and water of mice did not change significantly and its activity and defecation was normal.The fur looked smooth and bright.the weight of the mice gained naturally from20.017±2.0225g before the experiment to28.775±0.6552g. No mice was dead during the drug administration and all the mice survived and major organs functioned normally during the successive14day observation.ConclusionCLKG has obvious antiviral effects to influenza A virus H1N1, influenza A virus H3N2and influenza RP8srain and certain antiviral effects to parainfluenza, rhinovirus type14、 respiratory syncytial virus、 adenovirus type3and Coxsackievirus B3. The curable indicators of CLKG pills to those viruses are lower than ribavirin. CLKG could decrease the mortality and lung indexes of mice and the results are better than ribavirin.the actue toxiology test does not get LD50fo mice.No mice death and acut toxic manifestation occurred shows that CLKG pill is low toxic or nontoxic.The resuls above show that the components of CLKG pill are scientific.The pill has strong antiviral effects and therapeutic function towards prevalent cold caused by the invation of heat evils and epidemic toxins and it is clinically safe.