The Acute Toxicity Of 5 Kinds Of Extracts From Traditional Chinese Medicine (TCM) And Anti-HIV-1 Effect Of Its Chinese Herbal Formulas In Vitro

As research of Natural products against AIDS increasing, effecti ve anti-hiv drugs of screening from the rich resources of traditional Chinese medicine and further screening out active ingredients and a ctive site has become a hot spot of the current anti-aids drugs rese arch.traditional Chinese medicine extract can be recombination acco rding to their toxicity and combine with tonic kind of traditional Chinese medicine, which can better play the important role in anti-H IV effects, this compound preparations of Chinese medicine has become a new research point.Objective1、Study of the acute toxicity of traditional Chinese medicine Petroleum ether extract of Brucea javanica (No.1 extract), Co rtex Meliae (No.2 extract).Pharbitis nil choisy (No.3extract), chl oroform extract (No.4 extract) and N-butanol extract (No.5 extract) of Pharbitis nil choisy extract,which provides safety evaluation basis for the development of medicinal extract exploration and the i ntroduction of new breakthrough point in the research of extracts fro m compound preparations.2、Study of recombiniation of traditional Chinese medicine extrac t according to the toxicity, and combine with tonic kind of tradition al Chinese medicine, exploring how to play the more effectively role in resisting HIV-1Methods1、 Maximum tolerance doses(MTD) was obtained by use Maximum tole rance method to make acute toxicity test of No.1 extract, No.2 extra ct in KM mice, including observation of animal body weight, death, gen eral state and toxic signs for 14 consecutive day. The up and down m ethod (UDP)was used in acute toxicity test of No.3extract, No.4extrac t,, No.5 extract, body weight, death, general state and toxic signs f or 14 consecutive day was observed, LD50 value were be calculatedthrough AOT425StatPgm program and the corresponding 95% confidence interval.2、 By real time fluorescence quantitative methods (RT-PCR) compar ing concentration-response relationship of combination of different p roportion of extracts of traditional Chinese medicine.Result1、In Acute toxicity experiment, abnormal symptoms of No.1 extract and No.1 extract of animal death, animal strange behavior, reflection, hair, and so on, were not observed.Maximum tolerance dose (MTD) of No.1 extract and No.2extract were respectively 900g/kg/、21000g/kg. (Convert crude drugs of their MTD)No.3 extract, No.4extract, No.5 extract all showed toxic symptoms, median lethal dose (LD5o) respectively of No.3 extract, No.4extract, No.5 extract was3758mg/kg、325.5mg/kg、1098mg/kg, which suggested that the acute toxicity of three kinds of extracts in mice was No.4 extract >No.5 extract>No.3 extract.2. in Concentration-response relationship experiment of the combination of different proportion of extracts of traditional Chinese medicine (TCM), B combination according to the compatibility of acute toxicity was better in in inhibiting HIV than C combination by equal proportion of five kinds of extracts. No matter B combination according to the compatibility of acute toxicity or C combination by equal proportion of five kinds of extracts, there was no more obvious difference in inhibiting HIV comparing with single No.3 extract in dose 1 ug/ml-16 ug/ml, they just showed the same effect as the single extract, sometimes even lower effect.After adding suitable amount of epimedium and ginseng, HIVRNA expression in cells of BR combination and A combination had no significant difference comparing with No.3 extract’s in the concentration of 0.25 ug/ml~16ug/ml. HIVRNA expression in cells of AR combination had significant difference comparing with No.3 extract’s in the concentration of 4ug/ml~16ug/ml, which suggested that AR combination was better in inhibiting HIV than No.3 extract’s. Viral load in the supernatant of BR combination and AR combination had no significant difference comparing with No.3 extract’s in the concentration of lug/ml-16ug/ml. At the same time, HIVRNA expression in cells and viral load in the supernatant of AR combination showed the tendency of lower than 3 extract’s, BR combination, A combination, AR combination.ConclusionIn Acute toxicity experiment, Petroleum ether extract of Bruce a javanica (No.1 extract) and Cortex Meliae (No.2 extract) showed no obvious symptoms of the acute toxicityPetroleum ether extract of Pharbitis nil choisy (No.3extract) chloroform extract (No.4 extract) and N-butanol extract (No.5 extract) of Pharbitis nil choisy extract showed obvious symptoms of acute toxicity in different extent. B combination according to the compatibility of acute toxicity was better in in inhibiting HIV than C combination by equal proportion of five kinds of extracts, which suggested that B combination is reasonable. No matter B combination according to the compatibility of acute toxicity or C combination by equal proportion of five kinds of extracts, there was no more obvious difference in inhibiting HIV comparing with single No.3 extract in dose 1 ug/ml~16 ug/ml, which suggested that combination of extracts did not show the synergism. After adding suitable amount of epimedium and ginseng, HIVRNA expression in cells and viral load in the supernatant of AR combination showed the tendency of lower than 3 extract’s, BR combination, A combination, AR combination.According to the above results, we believe that the traditional Chinese medicine formula shouid be combined with multiple aspects of drug toxicity, detoxification and improving immunity.