| Gastrointestinal disease is the most common and important veterinary clinical disease, it seriously affect the healthy development of animal husbandry in China. Chinese veterinary medicine has paied a great attention to the animal gastrointestinal disease prevention and control, The long time clinical practice shows that using Chinese veterinary and the combination of Chinese and western veterinary could achieve good prevention and control effects on gastrointestinal disease of livestock and poultry. Guanyuzhixiesan were comprised by the geranium wilfordii, garden burnet, sophora alopecuroides, cumin and liquorice according to the basic theory of the traditional Chinese veterinary medicine. It had prevention and control effects on gastrointestinal disease of livestock and poultry. And Guanyuzhixiekeli is granulated dosage forms developed by Guanyuzhixiesan. Based on Guanyuzhixiesan, the acute toxicity and pharmacodynamics of it had been stuied, then the quality control method of Guanyuzhixiekeli had also been stuied. The specific results were as follow.1. Eight SPF Kunming mice had been chosen and then were randomly divided into the experimental and the control groups, and with four mice in each group. Each mice in the experimental group was given 0.2 m L/10 g Guanyuzhixiesan solution which concentration was 10 g/m L, and three times in 1 day, so, each mice was given Guanyuzhixiesan solution about 600 g crude drugs /kg in total in 1 day to peform the preliminary study of the acute toxicity, the control group was given the same amount of saline solution. After this, observed for 7 d, there had no obvious toxic reaction appeared. Twenty mice were chosen in the formal experiment, and divided into the experimental and the control groups, with ten mice in each group, then using the same method and dose of Guanyuzhixiesan and physiological saline with the preliminary study to give each group. Observing for 7 d, too. All the mice were alive and the group of mice which were given Guanyuzhixiesan had no significant difference between the control group in changes of weight and feed intake, and all mice had no visible to the naked eye visceral changes been observed after autopsy. However, this dose has been far in excess of the traditional Chinese medicine non-toxic dose standard(LD50>50 g/kg), this showed that there has no obvious acute toxicity reaction of Guanyuzhixiesan.2. Using folium sennae gavage combined with ice acetic acid enema method to duplicate rat ulcerative colitis model with spleen deficient, with general behavior of rats, routine blood, liver and colon tissue pathology, interleukin(IL)-1β, IL-6, substance P(SP), vasoactive intestinal peptide(VIP), malondialdehyde(MDA) and superoxide dismutase(SOD) as the evaluation index, the effects of Guanyuzhixiesan on ulcerative colitis of rat with spleen deficient were been studied. The results showed that compared with the self-healing group, the degree of diarrhea, the mental status, the diet, the fur color, the weight, the liver pathological changes and the colon pathological changes of the rats in the Guanyuzhixiesan group were significantly recover, the content of WBC and NE% in the blood, the IL-6, IL-1β and MDA in serum, the SP in colon tissue were significantly decrease(P<0.05), while the activity of SOD in colon tissue and the content of VIP in serum were significantly increase(P<0.05). It suggest that Guanyuzhixiesan has a very good therapeutic effects of the rats with ulcerative colitis. Its mechanism may be related to the inhibition of the secretion of IL-1β, IL-6 and SP, the reducing of the content of MDA, and the enhancement of the secretion of VIP and the activity of SOD.3. Quercetin and gallic acid were chosen as the reference substance, then the thin layer chromatography(TLC) and the high performance liquid chromatography(HPLC) were separately used to qualitative identification and determination the content of quercetin and gallic acid in the Guanyuzhixiekeli. TLC results showed that the spots of Guanyuzhixiekeli were clear and corresponding to the spots of quercetin and gallic acid standard substance, while the negative reference substance had no spots to correspondence to, these suggested that Guanyuzhixiekeli contain quercetin and gallic acid, and these TLC method could be used as the qualitative identification method of quercetin and gallic acid in the Guanyuzhixiekeli. HPLC results showed that there had good linear relationship with peak area when the concentration scope of quercetin and gallic acid were 4.86~38.88 μg/m L(r=0.99937) and 4.24~33.92 μg/m L(r=0.99913), and the content of quercetin and gallic acid wre 28.88 mg/kg(RSD=1.00) and 157.31 mg/kg(RSD=1.03), and these two method had high precision, good repeatability, stable samples within 24 h, high recovery rate after been investigated. So, they could be used as the content determination method of quercetin and gallic acid. In general, these method had high degree of separation and good reproducibility, they could be used as the quality control method of Guanyuzhixiekeli. |
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