The Function To Prevent Diarrhea And Safety Of Pulsatilla, Phellodendron And Their Compound Extract On Rats

According to Chinese medicine theory, Pulsatilla and Phellodendron as the main prescriptions of Chinese herbal medicine compound were designed. This article was conducted to study the inhibiting function on Escherichia coli in vitro and the effects on body weight, feed intake, diarrhea and intestinal tract bacterial communities of the rats infected by Escherichia coli of Chinese herbal medicine compound and its main prescriptions, to provide the basis of clinical application for Pulsatilla, Phellodendron and their compound extract through the maximal dose test.The bacteriostatic function of Pulsatilla, Phellodendron and their compound extract in vitro (Experiment 1)The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of Pulsatilla, Phellodendron, Pulsatilla:Phellodendron (1:1) and Chinese herbal medicine compound on Escherichia coli were measured by using double dilution method for three times, and made three repeats of each concentration. The results showed that each group of Chinese herbal medicine extracts all can inhibit the growth of E. coli, but the compound was the most obvious, MBC was 0.1250g/ml; the same effect of Pulsatilla 1 and Pulsatilla:Phellodendron (1:1), MBC was 0.2500g/ml; the effect of Phellodendron was not obvious, but showed resistance. So the inhibition effect of compound is not the simple addition of its components. The inhibiting effects of Phellodendron in compound was very obvious, it can effectively inhibit and kill the pathogen by synergy effect with other prescriptions.The function to prevent diarrhea of Pulsatilla, Phellodendron and their compound extract (Experiment 2)50 rats with 18-22g body weight were randomly divided into 5 groups:control and medicine groups(Pulsatilla, Phellodendron, Pulsatilla:Phellodendron (1:1) and Chinese herbal medicine compound group), each of which was assigned with 10 rats, half male and female, the pre-trial and testing period was 5 and 13 days. Rats were given extract by intragastric administration according to 4g crude drug/kg body weight at the same time everyday, the control was given the same dose distilled water. Each Rat was given 108 E. coli bacteria/ml by intraperitoneal injection at the 10th day to establish the model of diarrhea which caused by bacteria, and observe the effect of extract to diarrhea and loose stools rate. Collect the dejecta at the 13th day, DGGE analysis technology was used to monitor the structure change of intestinal bacteria. The results showed the change in body weight of control was the most obvious at the 10th day, were significantly different with the other groups (P＜0.05), and diarrhea was the most serious; each group of Chinese herbal medicine extract all can inhibit the diarrhea caused by E. coli, compared with control group, the compound group was the most obvious, the diarrhea times decreased by 51% (P＜0.05), were no significant difference between the other Chinese herbal medicine groups. DGGE profiles showed the number of DGGE bands and diversity index of compound group were significantly more than the other treatment groups (P＜0.05), were no significant difference between the other treatment groups (P>0.05); UPGMA clustering similarities analysis showed the degree of similarities of Chinese herbal medicine groups were all more than 63%, and are attributed to a cluster(except 5), but the degree of similarities of control were range from 56%-69%, individual differences was large. So Chinese herbal medicine can effectively reduce the diarrhea caused by E. coli on rat, improve the communites of intestinal tract bacterial and made the intestinal microecology more optimal and balance, so can ensure the relative stability of intestinal tract bacterial communities, in which the effect of compound extract was the most obvious.The safety of Pulsatilla, Phellodendron and their compound extract (Experi-ment 3)40 rats with 18-22g body weight were randomly divided into 4 groups: control and medicine group(Pulsatilla, Phellodendron and Chinese herbal medicine compound group), each of which was assigned with 10 rats, half male and female, adapted to the environment for 5 days, the maximal dose method was used in acute toxic experiment, and observe 7 days. The results showed:compared with control group, the body weight, feed intake, blood biochemical parameters and viscera/body weight ratio of Chinese herbal groups were not significantly different (P>0.05), in which rats were given extract twice in 6 hours according to 0.4ml/10g body weight by intragastric administration (to be equivalent to 136.8g crude drug/kg body weight), and their organs were all no obvious pathological change. So all extracts in this experiment were safe, non-toxic side effects to rat and high security for clinical use. They can be applied to production.In summary, the inhibiting effect of Pulsatilla, Phellodendron and their compound extract on Escherichia coli were all well. There are many antimicrobial constituents in compound extract, efficiently improve the communites of intestinal tract bacterial, increase the quantity of probiotics, decrease the quantity of probiotics of harmful bacteria, so can significantly reduce diarrhea caused by E. coli. And from the toxicity and side effect point of view, all extracts are safe, has a bright future for application in animal husbandry.