CN105769894A

CN105769894A - Applications of rutin in preparing medicines for preventing and treating influenza A

Info

Publication number: CN105769894A
Application number: CN201410769871.XA
Authority: CN
Inventors: 房学迅; 于淼; 王烨; 于大海
Original assignee: Individual
Current assignee: Individual
Priority date: 2014-12-15
Filing date: 2014-12-15
Publication date: 2016-07-20

Abstract

Te invention provides applications of rutin in preparing medicines for preventing and treating influenza A, belonging to the field of applications of traditional Chinese medicines. The technical scheme is as follows: firstly, neuraminidase liquid extracted from H1N1, H3N2 and H7N9 viruses and rutin are incubated in a 37 DEG C incubator for 30 min, then fluorogenic substrate buffer solution is added for carrying out detection, and the result shows that rutin has an inhibition effect for the neuraminidase of the three influenza viruses; then the effects of rutin in resisting the influenza A viruses in the cellular level are detected, and the result shows that rutin also can play prevention and treatment roles for the cells infected with the influenza A viruses; finally, the influences of rutin on a mouse influenza model are detected, the result shows that after rutin is added, the weight loss degree of mice infected by the influenza A viruses can be obviously inhibited, then the mice gradually recover to in the healthy state, and meanwhile, the lung indexes of the mice infected by the influenza can be obviously reduced.

Description

Rutin application in preventing and treating influenza A medicine

Technical field

The invention belongs to application in TCM field, be specifically related to rutin and suppress the application in influenza A virus medicament in preparation.

Background technology

Influenza (hereinafter referred to as influenza) is the acute viral respiratory infectious disease caused by the RNA viruses of myxovirus section, its infectiousness is strong, popular wide, sickness rate is high, the whole world nearly 5%～15% is infected every year.Myxovirus section divides 5 genus: influenza A virus genus, Influenza B virus genus, influenza virus C genus, Infectious salmon anaemia virus belong to and holder high soil Tobamovirus.At present, the most common being also easiest to of influenza A virus causes pandemics and area popular.

According to statistics, in 3 the large-scale influenzas broken out in 20th century, being to occur at the harm that " spanish influenza " of 1918 cause respectively the most serious, death toll is up to more than 50,000,000 people；The death toll that " Asia influenza " occurred in nineteen fifty-seven causes reaches 100～1,500,000 people；With the death that " Mao flu " that occur in nineteen sixty-eight causes 75～1,000,000 people.2013, having broken out a kind of novel type A avian influenza virus H7N9 in China, this virus only finds in the past in birds, is find that it can infect the mankind first specifically.Between for the moment, common people talk ' fowl ' complexion changed, the rapid atrophy of domestic fowl farming.The deputy to the National People's Congress, Nantong Affiliated Hospital of university Liu vice-president give one group of data: according to measuring and calculating, and within 2013, the whole industry loses 100,000,000,000 yuan, account for nearly the 20% of the output value.In January, 2014, industry lost 20,000,000,000 yuan, accounted for more than the 30% of the output value.Present middle-size and small-size poultry enterprise closes a business in a large number, and large-scale leading enterprise is by the underproduction 30%, Zhong Qin enterprise underproduction 30-50%.Some banks eliminate the loan to domestic fowl farming or force repaying.Many business capital chain interruptions, poultry had been absorbed in the edge of collapsing already.

Chinese medicine antiviral has the history of thousands of years, and China have accumulated a large amount of valuable theory and practice experience in utilizing Chinese herb prevention influenza.In China, natural resources of Chinese medicinal materials is very abundant, evident in efficacy by Chinese medicine influenza from ancient times to the present.But abroad, due in Chinese medicine effective ingredient fail to understand, action principle is unclear, so that the development of Chinese medicine is obstructed abroad.The research of present single medicinal material influenza has had many decades, has filtered out the Chinese herbal medicine much with resisiting influenza virus.So, we obtain the compound rutin with NA inhibitory activity of novelty from natural product, find suppression influenza NA with this, thus suppressing the novel drugs of influenza virus.

Summary of the invention

It is an object of the invention to provide rutin application in preparation preventing and treating influenza A virus medicament.Rutin can effectively suppress the activity of neuraminidase, thus suppressing influenza virus breeding in vivo, and then alleviates or cures flu victims.The neuraminidase of H1N1, H3N2 and H7N9 has been carried out the detection of external zymetology inhibition by compound rutin by the present invention, it has been found that its IC₅₀Respectively 102.17 ± 11.56,134.69 ± 13.59 and 104.17 ± 5.37 μm of ol/L, we again to these three influenza infection mdck cell and in addition this kind of compound carried out the mensuration of cell survival rate, it has been found that this compound all can improve the cell survival rate after viral infection.Finally the body weight change of influenza mouse model, fatality rate and Lung Exponent have been measured by we by this kind of compound rutin, it has been found that rutin can substantially alleviate the influenza virus symptom to mice.The present invention provides basis for exploitation with the new drug that neuraminidase is target spot, and the treatment being applied to Human Influenza for rutin provides reference.

Accompanying drawing explanation

Fig. 1 is the variable concentrations rutin inhibitory action to H1N1, H3N2 and H7N9 influenza neuraminidase.

Fig. 2 is after variable concentrations rutin and H1N1, H3N2 and H7N9 combined effect, the mixed liquor damaging action to mdck cell.

Fig. 3 is the therapeutical effect of the variable concentrations rutin mdck cell to having infected H1N1, H3N2 and H7N9.

Fig. 4 is the impact without the concentration rutin Mouse Weight on having infected H1N1 influenza virus.

Fig. 5 is the impact without the concentration rutin Mouse Weight on having infected H3N2 influenza virus.

Fig. 6 is the impact without the concentration rutin Mouse Weight on having infected H7N9 influenza virus.

Fig. 7 is to having infected the impact of virus load in the mouse lung of H1N1, H3N2 and H7N9 influenza virus without concentration rutin.

Detailed description of the invention

Below by way of the detailed description of the invention of example forms, the present invention is described in further detail.

Embodiment 1

We are with the H1N1(A/NewCaledonia/20/1999 containing influenza A virus), H3N2(A/Fujian/411/2002) neuraminidase with containing restructuring influenza A virus H7N9 (A/Anhui/1/2013) is prepared in pH6.5MES buffer (32.5mM), we join in reaction buffer by the mixture by the NA liquid of 49 μ L and with 1 μ L rutin of variable concentrations, 37 DEG C of incubators are placed 30min, it is subsequently adding L20 μM of MU-NANA substrate buffer solution of 50 μ, microplate reader detects, excitation wavelength is 360nm, transmitting wavelength is 450nm, detection duration 8min.Testing result is, the activity of H1N1, H3N2 and H7N9 influenza neuraminidase is suppressed concentration (IC during half by rutin₅₀) respectively 102.17 ± 11.56,134.69 ± 13.59 and 104.17 ± 5.37 μm of ol/L.

Embodiment 2

In order to detect the rutin influenza effect at cellular level, it is tested by we with process two kinds different: first by Concentraton gradient, rutin is diluted to variable concentrations, and I by above-mentioned solution and 100TCID₅₀H1N1, H3N2 and H7N9 influenza virus equal-volume hatches 2 hours, and cumulative volume is 200 μ L.Then mixed liquor is joined and mdck cell is hatched 2h；II by 100TCID₅₀H1N1, H3N2 and H7N9 influenza virus hatches 2 hours in mdck cell, add hatch 2 hours without the rutin of concentration.2 kinds of different disposal diluents all use maintenance medium (DMEM of 10 μ g/ml pancreatin).After 2 hours, change once above-mentioned maintenance medium and continue to cultivate 24 hours.Change the maintenance medium containing 2% serum, Continuous Observation 2～3 days after 24 hours, detect cytopathic effect with mtt assay.Above-mentioned experiment is all at 37 DEG C, 5%CO₂Cultivate under condition.

As shown in Figures 2 and 3, when different disposal, when adding rutin with influenza virus, along with the increase of rutin concentration, cell survival rate also increases as result.Illustrating that rutin can suppress influenza virus, protection mdck cell is from viral subversive, and protective effect presents concentration dependent also with the increase of rutin concentration.

Embodiment 3

In order to detect rutin protective effect in mouse influenza models, we adopt 50 μ L10LD₅₀H1N1, H3N2 and H7N9 influenza virus collunarium infecting mouse.Experiment mice is randomly divided into 6 groups, respectively NC group；10LD₅₀Group；10LD₅₀+ 50mg/kgTamiflu group；The basic, normal, high dosage group of rutin: 10LD₅₀+ 50,100,200mg/kg rutin.Often group 10, virus attack starts administration for first 1 day, all adopts the mode of gastric infusion, every day 2 times, continuous 7 days, every mouse stomach 0.2ml.Record mouse survival state, body weight change every day.Then when 15 days, put to death mice, record lung weight, lung tissue is ground and adds PBS, measure its NA activity afterwards.

Result as shown in Figure 4, Figure 5 and Figure 6, illustrates that what rutin can suppress influenza infection Mouse Weight alleviates degree, improves the survival condition of mice, make mice be gradually restored to health level.

Result is as shown in the table, illustrates that the rutin of variable concentrations can significantly reduce the Lung Exponent of influenza mice.

Result above all illustrates, the mice infecting influenza can be played the effect for the treatment of by rutin, makes mice be gradually restored to health level.

Claims

1. compound rutin application in preventing and treating influenza A medicine.

2. as in claim 1 rutin with the application of Oral administration.

3. as in claim 1 rutin with the application of drug administration by injection mode.

4. as in claim 1 rutin with the application of suction of spraying.

5. as in claim 1 rutin using as single component preventing and treating influenza A medicine application.

6. as in claim 1 rutin using as a kind of composition in compound medicine preventing and treating influenza A medicine application.