ABSTRACT
<p><b>OBJECTIVE</b>Modeling HAdV-3 infect Hep-2 cells in vitro. The effect of realgar nanoparticles on the expression of HAdV-3 is detected by using fluorescent quantitative PCR.</p><p><b>METHODS</b>The experiment is divided into four groups: Hep-2 cells control group, HAdV-3 virus control group, realgar nanoparticle group and ribavirin group. In order to detect HAdV-3 viral load, add realgar nanoparticles and ribavirin in vitro and remain that vitro for 24 hours when HAdV-3 has infected Hep-2 cells, extract total DNA of Hep-2 cells infected by HAdV-3, and establish Real-time PCR reaction system of every experimental groups.</p><p><b>RESULT</b>The Hep-2 cells group has no amplification curve, the Ct value is greater than 35, which illustrate HAdV-3 pathogen detection is negative. However, realgar nanoparticles group, ribavirin group and the HAdV-3 group have amplification curve, the Ct values are 29.30 +/- 0.08, 33.05 +/- 1.29, 26.01 +/- 0.25 respectively, which illustrate HAdV-3 pathogen detection is positive. The viral copy amount of the adenovirus group(66 699 932 +/- 23.85) is more than that of realgar nanoparticles group (912 435.44 +/- 16.57), and much greater than that of ribavirin group (459 124.84 +/- 12.82) (P < 0.05).</p><p><b>CONCLUSION</b>The model of Hep-2 cell infected by HAdV-3 is reliable. The method of quantitative PCR is sensitive and specific. Realgar nanoparticles have a certain inhibition role for adenovirus nucleic acid replication.</p>
Subject(s)
Humans , Adenoviridae Infections , Virology , Adenoviruses, Human , Genetics , Physiology , Arsenicals , Chemistry , Pharmacology , Gene Expression Regulation, Viral , Hep G2 Cells , Nanoparticles , Chemistry , Sulfides , Chemistry , Pharmacology , Virus ReplicationABSTRACT
<p><b>OBJECTIVE</b>In order to screen out a certain kind of traditional medicine which has a better role in immune regulatory, the influence of representatives of heat clearing and detoxicating herb on inflammatory cytokines protein expression of mice lung homogenate infected by FM1 have been observed.</p><p><b>METHOD</b>Modeling mice infected by FM1. On the first, third, fifth and seventh day after FM1 infection, tumor necrosis factor-alpha (TNF-alpha), interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-10 (IL-10), and gamma-interferon (IFN-gamma) expression in mice lung homogenate of normal control group, model control group, scutellari group, isatidis group, pulsatilla group, polygonum cuspidatum group and oldenlandia group have been tested by ELISA method.</p><p><b>RESULT</b>The expression of TNF-alpha, IL-6, IFN-gamma and IL-10 in mice lung homogenate reaches its peak on the third day after FM1 infection, significantly higher than the control group (P < 0.05). Scutellari and isatidis are two representatives of heat clearing and detoxicating herb, which can decrease the expression of TNF-alpha, IL-6 and IL-1 and increase the expression of IL-10, IFN-gamma. The effect are more pronounced and statistically significant (P < 0.05) on the third and fifth day after infection, pulsatilla, polygonum cuspidatum and oldenlandia can also regulate the inflammatory cytokines, but the effect are not so obvious as scutellari and isatidis.</p><p><b>CONCLUSION</b>Scutellari and isatidis, two representatives of heat clearing and detoxicating herb, have a good intervention on immune damage caused by influenza virus through adjusting the balance of inflammatory cytokines and anti-inflammatory cytokines.</p>
Subject(s)
Animals , Chick Embryo , Humans , Male , Mice , Cytokines , Genetics , Allergy and Immunology , Disease Models, Animal , Drugs, Chinese Herbal , Therapeutic Uses , Gene Expression , Inflammation Mediators , Allergy and Immunology , Influenza A virus , Physiology , Influenza, Human , Drug Therapy , Genetics , Allergy and Immunology , Virology , Lung , Allergy and Immunology , Virology , Mice, Inbred BALB CABSTRACT
This study was to establish a model to explore anti- RSV effect of different administration method of Chinese medicine realgar on respiratory syncytial virus type A (RSV-A) replication in Hep-2 cells. Using high-energy ball milling with distilled water to prepare realgar nanoparticles,the concentration of nanometer realgar was tested by molybdenum blue staining method and the size of realgar nanoparticles was tested on Nano Series. Cell culture with ribavirin as a positive control was applied to observe the effect of anti-respiratory syncytial virus type A replication through prevention, treatment or direct inactivation of three different drug administration methods. Realgar nano-particles was found to be a potential inhibitor of RSV-A in a concentration-dependent manner with the median toxic concentration(TC50) of 0.649 microg/mL in Hep-2 cell culture. The median inhibition concentration (IC50) was 0.20 microg/mL when drug was added before virus infection. The IC50 was 0.13 microg/mL when drug was added after virus infection,and it was 0.16 microg/mL when the drug was mixed with virus and added. The therapeutic index (TI) was 3.18, 4.99 and 4.11, respectively. The results showed realgar nanoparticles could inhibit the replication of the RSV and inactivate the RSV in vitro.
Subject(s)
Arsenicals , Pharmacology , Nanoparticles , Respiratory Syncytial Viruses , Physiology , Sulfides , Pharmacology , Virus ReplicationABSTRACT
<p><b>OBJECTIVE</b>This study was to establish a model that adenovirus type 3 (HAdV-3) infected on Hep-2 cell in order to explore anti-adenovirus3 (HAdV-3) effect of Chinese medicine realgar in vitro.</p><p><b>METHOD</b>Use high-energy ball milling with distilled water to prepare realgar nanoparticles. The concentration of nanometer realgar was tested by molybdenum blue staining method and realgar nanoparticles' particle size was tested on Nano Series. The technique of cell culture with ribavirin as positiv control was to observe anti-adenovirus effect through prevention, treatment and direct inactivation of three kinds of drug delivery.</p><p><b>RESULT</b>This drug was found to be a potential inhibitor of HAdV-3 in a concentration-dependent manner with the median toxic concentration (TC50) of 0.649 microg/ml in Hep-2 Cell culture. The median inhibition concentration (IC50) was 0.255 microg/ml when drug was added before infection. The IC50 was 0.142 microg/ml when drug was added after virus infection, and it was 0.117 microg/ml as the drug was added after it mixed with virus. The therapeutic index (TI) was 2.55, 4.57 and 5.55 respectively.</p><p><b>CONCLUSION</b>The direct inactivation effect of realgar nanoparticles is the most evident in three drug deliveries manner with the same concentration in vitro.</p>