| HIVl (human immunodeficiency virus) is the pathogen of AIDS. HIV mainly invasion CD4+ T lymphocytes and macrophages. In the previous stages ol infection, immune systems can clear out replicated virus in time and without any symptom. With the progression of illness, the amount of CD4+ T cells decrease gradually and immune systems trends to exhaustion. When the number of CD4+ T cells is to 200/ml , infected persons often died of opportunistic infections and tumors. From 1980', epdemic of HIVl spread worldwide rapidly and 95% of infectors are ii; the developing country. It is reported that the number of infectcrs in China amounted to more than 850 thousands. Anti HIV therapy is the focus in global studying. Up to date, there are not safetyeffective vaccines and there are no drugs that be able to clear out virus thoroughly. For synthetic compounds have the shortcomings of resistance and seriously toxicity and side effect ,and numerous persons that infected HIV1 can't be able to afford expensive therapy.Herbs have been used for centuries in china and continue to be used extensively to treat a variety of illnesses. Clinical study has showed some interests in the anti-HIV activity of traditional medical herbs. In 1984, four hemophilia patients were diagnosed infecting HIV1 in Zhejiang, China that they had ever transfused import concentrate V III factor. Reported clinical information showed that the condition of HIV infectors by transfuse transmission would deteriorate rapidly and only survive 2 to 5 years compared with infection by the other transmission ways. These four HIV infectors had been received Chinese medicine regimen-AIKANG for therapy. The course of disease went slowly and one survived 18 years.There're eight kinds of herbs in AIKANG regimen, include glycyrrhiza, viola yedoensis, Makino, coptis and Aogelica, et al. According to some rules, we got extractors from these herbs and compounded these herbs and then , study their inhibitory active to HIV in vitro. Of course, the previous study had proved that the clinic oral dosage is safety and the mice had even able to taken dose by oral 50 to 60 times than human.There are 14 kinds of extractors and 22 kinds of compounds. These extractors and compounds were tested for their ability to inhibit HIV1 replication and adsorption in HIV-1 MT4 culture system in vitro. MT4, An HTLV-1 transformed T4-cell line, whichwas previously proved to be highly susceptible to and permissive for, HIV infection, served as the target cell line.Inhibiting duplication experimental method: first, the drugs are diluted according to requirement concentration by fresh culture medium in 96 well plate, then add into MT4 cell (6*105/ml, inoculated HIV-1 of titer 200TCID) 100|al per pore respectively , and no drugs uninfected control group (without HIV-1) and infection control group (added HIV-1) be thought about, which are cultured six days at 37癈. changing culture medium that including the same as drugs concentration per three days. The culture supernate is collected at the sixth day and measured P24 with ARKSU kit produced by Germany. So the inhibition effect (duplication inhibition rate) is calculated. The inhibition rate of CPE is also calculated at the same time by measuring the survival rate of cell with MTT method. P24 is specific nuclear corn protein of HIV viron, which is the main evaluation indication.Inhibitory adsorption experimental method: the drugs are diluted according to requirement concentration by fresh culture medium in 96 well plate, then add into MT4 cell (Ixl06/l)and HIV1 ( 200TCID) respectively 50 jil per pore, then did as same as above. Inhibition of the HIV introduced cytopathic effect(CPE) and quantity of P24 production as the end point.The results showed that an overwhelming majority of extractors have active to inhibit HIV1 induced CPE. In all extractors and compounds, some are able to inhibit HIV adsorption to MT4 cell and extractor H and compound 3 characterize by stable active. To analysis further the effect mechanism, we mixed H or com... |
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