Study On Lymphocyte Immunity Enhancement And Inhibition Of AMDV Proliferation Of Qizhushecao Compound

Mink is both a fur animal and a medicinal animal.Mink heart,mink whip,and mink liver can be used as medicine.Mink heart is the Principle Drug of Lixin Wan,which has a unique effect on the treatment of rheumatic heart disease.Dingxin Dan is a Chinese herbal compound preparation with Mink heart as the Principle Drug,which is clinically used for tachycardia,arrhythmia,heart failure,hypertension,and so on.Mink whip has a therapeutic effect on impotence.Mink liver has a therapeutic effect on night blindness.As people’s health awareness increases,the demand for mink hearts also increases year by year.Therefore,it is of great significance to ensure the adequate supply of mink hearts.The infection of Aleutian mink disease virus(AMDV)is one of the main causes of mink death,which seriously affects the supply of mink heart.It is imperative to look for anti-viral drugs.Many Chinese medicines have the function of enhancing immunity and anti-virus,and they are safe to use.Therefore,it is of great significance for the supply of mink heart to screen pure Chinese herbal compound that has immune enhancement effect and can inhibit the virus.This study is based on the principles of traditional Chinese medicine prescriptions,such as Yupingfeng San,Sijunzi Tang,Buzhongyiqi Tang,then add or subtract the ingredients,Astragalus or Codonopsis is the Principle Drug,Atractylodes macrocephala is the Minister Drug.At the same time,the Chinese veterinary medicine should be inexpensive.So Hedyotis diffusa or Eucommia ulmoides was added as Assistant Drug and Envoy Drug to form three compounds.A screening study of the three compounds was conducted on the effect of stimulating the proliferation of mouse splenic lymphocytes in vitro.As a result,a compound Chinese traditional medicine C(Qizhushecao Compound)with good stimulation effect on mouse splenic lymphocytes was obtained initially.Next,taking the direct proliferation effect of mouse splenic lymphocytes in vitro and the synergistic effect of Con A and LPS as indicators,using the uniform design method,the best ratio of Astragalus,Atractylodes,and Hedyotis diffusa in Qizhushecao compound was screened.The results showed that F4 could not only directly promote the proliferation of lymphocyte,but also synergistically with Con A and LPS,so Qizhushecao compound F4 was chosen as the candidate.Further,the effect of Qizhushecao compound F4 on the secretion of cytokines by mouse splenic lymphocytes and the proliferation of mink splenic lymphocytes was tested.The results show that Qizhushecao compound F4 can significantly promote the secretion of IL-2,IL-4,IL-12 and IFN-γ in mouse spleen lymphocytes,and promote the proliferation of mink spleen lymphocytes.In order to investigate whether the Qizhushecao compound F4 has an inhibitory effect onAMDV proliferation,q-PCR method was used to detect its inhibitory effect on AMDV proliferation on CrFK cells cultured in vitro,the results showed that when the drug concentration was 12 mg/m L and 6 mg/m L,the inhibitory effect was more obvious,and the AMDV gene copy number was significantly reduced compared with the virus control group(P<0.05),indicating that the Qizhushecao compound F4 has the ability to inhibit the proliferation of AMDV on CrFK cells.Further,the Qizhushecao compound F4 was fed to AMDV positive mink in three doses of high,middle and low,respectively,and the copy number of AMDV gene,the content of γ-globulin and the content of immune complex in serum samples before and after administration were detected.The results showed that the high-dose group and the middle-dose group of Qizhushecao compound F4 can significantly reduce the viral gene copy number,the content of γ-globulin and the content of immune complex in mink serum(P <0.01),and the low-dose group can reduce the copy number of viral gene and the content of immune complex(P <0.05),it indicates that the Qizhushecao compound F4 can inhibit the proliferation of AMDV in mink.The above results illustrated that the Qizhushecao compound F4 composed of Astragalus,Atractylodes macrocephala,and Hedyotis diffusa will have a better immune enhancement effect and can inhibit the proliferation of AMDV in CrFK cells and mink.