Study On The Inhibition Of Ganluxiaodu Dan Decoction On CoxA16 Virus In Vitro Drug Efficacy

Objective:To investigate the inhibiting effect of Ganluxiaodu Dan Decoction on Cox Al6 virus in cultured Hela cells and provide some experimental basis for illustrating the mechanism of the effect of Ganluxiaodu Dan Decoction.Method:To observe the inhibitory effect of CoxA16 virus in cultured HeLa cells,Ganlu Xiaodu Dan of Chinese medicine decoction, drug containing serum and ribavirin injection are added to human cervical carcinoma(Hela) cells respectively through direct inhibition of the virus, therapeutic inhibition of virus and intervention of anti-virus experiment. At the same time, the control group,the normal control group and ribavirin injection control group are set up;The level of INF-? in He La cell supernatant was detected by enzyme linked immunosorbent assay(ELISA);Transmission electron microscopy is used to observe the ultrastructure of He La cells in the 2h, 8h, 16 h and 24 h after the addition of Ganluxiaodu Dan Decoction.Results:(1)according to the Reed-Muench method to calculate TCID50,the result is10-5.361/ml;(2)The safe concentration of Ganluxiaodu Dan Decoction of Traditional Chinese Mdicine is 12.5mg/ml, the Ribavirin Injection is 12.5mg/ml.(3)Therapeutic effects of Ganluxiaodu Dan Decoction of Chinese Medicine on Cox A16 virus is the strongest, followed by the direct inhibition of Cox A16 virus test. The OD value of two sets of experiments in the drug group and virus control group have a significant difference(P < 0.05), The OD values of Ganluxiaodu Dan Decoction group and medicated serum group have a significant difference(p<0.05), compared witn Ganluxiaodu Dan Decoction and Ribavirin Injection group, significant difference is appearing(p<0.05), there is no significant difference between the Ribavirin Injection group and Ganluxiaodu Dan medicated serum group.The inhibition to Cox A16 virus of Ganlu Xiaodu Dan is relatively weak in intervention of anti-virus experiment. And there is no significant difference in the effect of anti Cox A16 virus in each drug group.(4)In detecting the level of INF-? in He La cell supernatant,the result is content of drug treated group > virus control group > normal cell control content.There has a statistically significance between Ganlu Xiaodu Dan Decoction of Traditional Chinese Medicine and drug containing serum group in the content of INF-?(P <0.05), the content of Ganlu Xiaodu Dan Decoction of Chinese Medicine and Ribavirin supernatant group are statistically significant(P < 0.05).but Ganlu Xiaodu Dan Decoction of Traditional Chinese Group and Ribavirin supernatant group had no significant difference.(5) The normal He La cell has microvilli on surface, no holes under electron microscope.The cell membrane began sinking after Cox A16 virus infected He La cells 2 hours;there are a number of virus particles in a spot of heterogeneous soluble enzyme after Cox A16 virus infected He La cells 8 hours;the number of heterogeneous macrophages are increasing after Cox A16 virus infected He La cells 16 hours and 24 hours.After adding Ganluxiaodu Dan Decoction 2hours,He La cell membrane is invaginat unconspicuous. After 8 hours treatment,there is no obvious white cytoplasm vacuoles in He La cells, mitochondria and endoplasmic reticulum swelling, cytoplasm began to shrink.The number of cytoplasmic endome of lysosomal macrophages is more than the model group after 16 hours treatment,it is wrapping and swallowing the particulate matter.The cytoplasm of lysosomal macrophages in cell disappearing after 24 hours treatment and the cell is recovering.Conclusions:1.The inhibiting effect of Ganluxiaodu Dan Decoction on Cox Al6 virus is strongest in therapeutic inhibition of virus experiment.followed by direct inhibition of virus experiment;Ganluxiaodu Dan Decoction inhibit Cox A16 virus is relatively weak in intervention of anti-virus experiment.2.Ganluxiaodu Dan Decoction block the biosynthesis of Cox A16 virus in He La cells by secreting INF-?,thereby it achieve indirect antiviral effect.