| This study is aimed at developing safe, effective anti-inflammatory Chinese herbal medicine prescription, which contains herbal medicine with such activities as heat-clearing, detoxifying, and promoting blood circulation, and primarilytreats the inflammation caused by biological factors including bacteria, viruses,and endotoxins. The prescription combination is designed on the basis of a dialectical research on the basic symptoms of the pathological feature in the inflammatory process, according to two therapeutic principles, including "yingxiefenzheng"of the febrile disease and "heat-clearing and blood-cooling, detoxifying" for treatment. The preparation of appropriate formulation is directed by a pharmacological indicator, the anti-inflammatory activity. Based on previous work, we use the uniform design method to perform studies on the decomposed recipes of the prescription, in order to improve the screening and design of the prescription combination. Besides, we evaluate the safety and efficacy of the prescription,establish the quality control standard, and determine the content of the index components at a preliminary level.The prescription under study is composed of 5 kinds of Chineseherbal medicine: artemisinin, gardenia, honeysuckle, salvia miltiorrhiza, and sarcandra glabra. The active ingredients are extracted and made into liquid medicine with a concentration of 1g/mL(crude drug). The liquid medicine is then mixed with dextrin-starch (1:2),before wet granulation is performed to prepare the compound granules, taking ethanol (95%) as a binder. Then, we investigate the micromeritic properties of the granules, and measure the critical relative humidity. Next, we employ the thin-layer chromatography (TLC) to identify the main active composition of the compound granules. We find that isofraxidin and geniposide can be identified successfully, when we select a solvent system of toluene-ethyl acetate-formic acid (9:4:1) and ethylacetate-acetone-formic acid-water (5:5:1:1) as the mobile phase. Finally, we use the high performance liquid chromatography(HPLC) to carry out quantitative analysis of the compound medicine,and establish the analytical method for two kinds of active ingredients artemisinin and isofraxidin. These results pave the way for the development of the prescription, as well as the establishment of the quality standard. the method of determination of was established and the above-mentioned 2 components were determined by HPLC. All these provide scientific basis for the development of compound preparation and the process for the modernizationof traditional Chinese medicine to provide the feasibility.Next, we investigate the anti-inflammatory effects of the prescription, by monitoring the auricle tumefaction caused by xylene, and the peritoneal capillary permeability increase induced by acetic acid in mice. It shows that the prescription can not only largely reduce the degree of tumefaction in auricle, but also suppress the capillary permeability increase. There is no significant difference (P<0.05) between the experimental group and the dexamethasone group (positive control group).To explore the safety of the prescription, we determine the maximum tolerable dose in mice. It shows that the difference between the treated and the blank group is not remarkable (P<0.05), regarding the mice weight and food intake.The maximum tolerance dose of the compound granules in mice is 0.403g/20g,which is 144 times more than the oral dose of human. These results indicate that the prescription is perfectly safe for human body.In addition, we investigate the effect of the compound granules on the artificial mice endotoxemia. The pathological model of endotoxemia in mice is established successfully by intraperitoneal injection of Escherichia coli lipopolysaccharide (LPS). It shows that the mortality rate of the LPS positive control group reaches 78.57% in 72 hours, while that of the group treated with dexamethasone(DEX) sodium phosphate injection is 28.57%. The difference between the LPS group and the DEX group is remarkable (P<0.05). In the experiment, themortality rate in the moderate dose group is 30.85%, which is higher than that of the DEX group, while significantly lower than that of the LPS group. There is significant difference (P<0.05) between the moderate dose group and the LPS group.To further explore the mechanism of the anti-endotoxin effect, we determine the content of TNF-a and IL-6 in serum,and the content of eNOS in liver. It shows that the content of TNF-a and IL-6 in serum of the moderate dose group is significantly reduced, the degree of which is comparable with that of the DEX group. The content of eNOS in liver of the LPS group increases at 10 hours after LPS injection, and returns to normal after 72 hours. In contrast, the content of eNOS of both the treated group and the DEX control group reaches peak value at 12 hours, and decreases sharply after 24 hours. We find that there is no remarkable difference between the treated group with moderate dose and the DEX control group. This study investigates preliminarily the mechanism of theanti-endotoxin effect of the compound granules that are made of Chinese herbal medicine, and will provide a new strategy for the treatment of endotoxemia.To explore the safety of the prescription, we determine the maximum tolerable dose in mice. It shows that the difference between the treated and the blank group is not remarkable (P<0.05), regarding the mice weight and food intake.The maximum tolerance dose of the compound granules in mice is 0.403g/20g,which is 144 times more than the oral dose of human. These results indicate that the prescription is perfectly safe for human body. |
PDF Full Text Request