| 1 Objective To change the traditional Mongolian medicine–Naru3 watered pills into Babu Plaster, that is to change the oral agent into percutaneous agent; to probe the characteristics and mechanism of the skin penetration of Naru-3 Babu Plaster, compare the difference in the blood concentrations of the two kinds of preparation, observe the effects of Naru-3 Babu Plaster and study the mechanism of Naru-3 Babu Plaster ameliorating the of rheumatoid arthritis (RA) and curing RA. 2 Methods 2.1 To prepare the extracts of kusnezoff monkshoot root by decoction and separating with macroporous resins, prepare long pepper extract by extracting with ethanol, prepare medicine terminalia fruie extract with decoction and filtration. 2.2 To determine the content of aconitine and meaconitine in the extracts of kusnezoff monkshoot root and chavicin in long pepper extract by high performance liquid symptoms chromatograpy (HPLC). 2.3 To collect the penetrated liquid including the extracts of kusnezoff monkshoot root and long pepper extract by method of modified Frands diffusing pond and determine the volume of the penetrated aconitine, meaconitine and chavicin by HPLC, calculate the constant of the velocity of the skin penetration. To collect the penetrated liquid of Naru-3 Babu Plaster by method of modified Frands diffusing pond and determined the velocity of the skin penetration of aconitine, meaconitine and chavicin by HPLC. 2.4 Getting blood from the carotid artery of rabbit, determine the volume of the penetrated aconitine, meaconitine and chavicin in the blood by HPLC after abstracting with ether. According to the result, observe the difference in the blood concentrations between the oral agent-Naru3 watered pills and the percutaneous agent-Naru-3 Babu Plaster. 2.5 Making aduvant arthritis(AA) model in rats with complete freund's adjuvant (CFA), observe the Anti-inflammatory effect according to the level of the swelling degree of metatarsus, the anti-swelling rate, the arthritis exponent, the secondary swelling, the erythema in the ear, the occurrence rate of nod on tail and the pathologic change in the ankle joint. 2.6 To determine the the levels of interleukin 1(IL-1), tumor necrosis factor α (TNFα) and prostaglandin E2 (PGE2) of the blood serum of the mice by enzyme - linked immunoadsordent assay (ELISA). 2.7 To observe the analgesia degree by the acetic acid induced body twisting in mice and measured the pain threshold by the hot-plate and study the analgesia effect of Naru-3 Babu Plaster. 3 Results 3.1 Part one Preparation of the traditional Mongolian medicine–Naru-3 Babu Plaster and the study on the characteristics of the skin penetration of Naru-3 Babu Plaster 3.1.1 To prepare the extracts of kusnezoff monkshoot root, long pepper extract and medicine terminalia fruie extract by decoction, separating with macroporous resins and extracting with ethanol. To determine the main contents of the extracts of kusnezoff monkshoot root and long pepper extract by HPLC. The results is that there are 3.9470mg meaconitine and 0.6898mg aconitine in 1g extracts of kusnezoff monkshoot root and 73.8239mg chavicin in 1g long pepper extract. 3.1.2 To determine the velocity of the skin penetration of aconitine, meaconitine and chavicin by Frands diffusing pond and HPLC. The results is that the velocity of the skin penetration of aconitine and meaconitine of high,middle and low concentration is 78.8052μg/cm2·h, 35.7333μg/cm2·h, 39.7052μg/cm2·h and 18.3909μg/cm2·h, 15.9628μg/cm2·h, 13.5627μg/cm2·h respectively, and the velocity of the skin penetration of chavicin of high, middle and low concentration is 265.5747μg/cm2·h, 208.2002μg/cm2·h and 5923μg/cm2·h respectively. 3.1.3 To compare the velocity of the skin penetration of meaconitine and chavicin under the condition that the concentration of the chemical preparations of promoting penetration-azone and propylene glycol was 2 percent, 4 percent and 6 percent respectivelyhe. The results is that the velocity of meaconitine is the biggest when the c... |
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