| PrefaceVas reaction is the central link of pathology on traumatic synovitis. It is effective and assured for the external Chinese traditional medicine to treat the disease. But the external preparations of Chinese traditional medicine are losing the competition power in the market because of some reasons such as antique preparation technics, unreasonable using of matrix, numerous and jumbled drugs in the compound, difficult investigation on the mechanism of transdermal absorbing and so on. Thus, it has been an important problem of T.C.M. modernization to develop an external preparation of Chinese traditional medicine which can possess the prominent and persistent effects and convenient using method. Transdermal drug delivery system(TDDS)is one of the modern external drug preparations, which can cure the whole body by way of the skin. And the TDDS is the center of the third era of exploitation research about drug preparation because it can reduce the toxicity and side-effect of drugs, advance the curative effect and decrease the administration times. The gelata, one of the preparation of TDDS, is a single, muddle or emulsion latex thick liquid or semisolid preparation made of the drugs and the pharmaceutical necessities which can become the gelata. The gelata preparation possesses the excellence of easy manufacture method andcomfortable using feeling.Doctors of the Affiliated Hospital of Liaoning University of T.C.M. have been treated traumatic synovitis using a proved recipe which contains fourstamen stephania root, with the effect of causing diuresis and detumescence, and cortex phellodendri, with the effect of clearing away heat and promoting diuresis. And the objectives of the present study were to study on pharmacodynamics and pharmacology of the T.C.M. compound gelata, containing the two drugs only, developed through appling the modern pharmacy technology and to provide experiment foundation for clinical application.Material and Methods1. AnimalsWistar rat, weighing 130 to 170g, and Kunming mouse, weighing 18 to 22g, from the Animal Study Center of the Liaoning University of T.C.M. were used.2. Instruments and reagentsfangjihuangbai gelata(self-made), votalin emulgel(made in Beijing NovatisPharmacy Limited Company), Carrageenan(Sigma Company), corneal trephine,electronic balance. Evans blue , acetic acid ,croton oil mixture corneal trephine, etc.(offered by T.C.M pharmacology test center of the Liaoning University of T.C.M.) ,751 spectrophotometer(shanghai analytical instrument main factory).3. Methods(1) Hot-plate test: In this test which measures analgesic activity, female mice were placed in the hot-plate water bath at 55±0.5℃, forecast the pain threshold through looking upon the mice's laping hind leg as the ache reaction guideline, regarded as qualified one whose pain threshold was belong to the range of 5～30 second. 40 mice with qualified pain threshold were distributed to 5 groups at random: excipient comparison group with the same dose(2g excipients /kg), positive control group with futalin cream (0.4g.kg—1), low dose group with fangjihuangbai gelata(0.04g crude drugs/20g); middle dose group with fangjihuangbai gelata(0.08g crude drugs/20g); high dose group with fangjihuangbai gelata(0.12g crude drugs/20g). the mice's back were daubed the drug with 1 cm2 area, one time in every day, continued 7days. At the 7th day, the mice were separately mensurated the pain threshold at 0.5h, 1h, 1.5h after daubed the gelata.(2) Writhing test: 40 health mice, weighing 18 to 22g, were prepared with the same numbers in different sexes. The grouping and daubing drug methods were as the same as the hot-plate test. Every mouse was intraperitoneal inject 0.2ml/20g 0.6% acetic acid at the 7th day. They were accurately recorded that the mice appeared the writhing reaction for the first time(delitescence) and the writhing times within 15mins. The drug's analgesic percentages were calculated.(3) The test affecting the permeability of blood capillary in mice: The mice were injected 0.1ml/20g 1% Evans blue in caudal vein, while intraperitoneal injected 0.2ml/20g 0.6% acetic acid in the 8th day. Then, the mice were executed by pulling apart their heads after 20mins, cut their abdomen, flushed the abdomen 3 times with 5ml distilled water, extracted 4ml the flushing liquid with straw, centrifugated 5min with 1000r/min and determined the figure of OD of 3ml supernatant at 590nm with 751 spectrophotometer.(4) Ear-swelling induced with croton oil test: 40 health male mice, weighing 18 to 22g, were grouped and daubed drug as that of the hot-plate test. At the 8th day, the mice were daubed around the right auricle with the croton oil mixture confected on the same day, 0.05ml for every one, executed by pulling apart cervical spine after 30mins from being caused inflammation. Then, the left and right auricles in mice were cut by corneal trephine with 8mm diameter and were weighed respectively by electronic balance. At last, the ear-swelling percentages were calculated.(5) Migrated leukocytes test in rat: 40 male Wistar rat, weighing 130 to 170g, were distributed to 5 groups at random: excipient comparison group with the same dose(0.3g excipients /150g), positive control group with futalin cream (0.4g.kg—1), low dose group with fangjihuangbai gelata(0.3g crude drugs/150g); middle dose group with fangjihuangbai gelata(0.6g crude drugs/150g); high dose group with fangjihuangbai gelata(0.9g crude drugs/150g). The rat were moulted on the back ,with 4 cm2 area, daubed the gelata on the part, once a day, continued to 7 days. At the, 8th day, the rat were hocused with aether, injected 1% Carrageenan (25mg.kg—1) in the left thorax. Then, the rat were executed after 4 hours, cut open the thorax, extracted 20μl thorax effusion liquid to 0.38ml leucocyte diluted liquid. At last, the leucocyte number in the liquid were counted by the leucocyte-count method.(6)reduce the content of PGE2 in inflammatory tissue of rat foot At the 8th day, every rat's right foot was injected 1 % Carrageenan after daubed the gelata 30 mins.Then the rat was excuted after 4 hours. The inflammatory foot was cut upper ankle 5mm and weighted. And the foot was put into normal saline for 1 hour, pick up the foot and centrifugate the liquid ,take supernatant .0.1 ml,add 0.5mol·L—1 KOH-CH2O liquid 2ml, water bath 20min at 50℃,and put formaldehyde solution 5ml in it , determinate the absorbance at 278nm with the 751 spectrophotometer(OD).(7) Statistical analysisData are reported as means±SEM. Data were analyzed by Dunnet -t test method.Results1. Fangjihuangbai gelata can advance the pain threshold, prolong the ache delitescence and reduce the writhing times of the mice. And the experimental results have the statistics significance.2. Fangjihuangbai gelata can reduce the permeability of blood capillary, decrease the phlogistic medium's effusion and control the auricle tumefy of the mice induced by croton oil(high and middle dose)and control the auricle tumefy of the mice.3. The phlogistic reaction induced by carrageenan in mice in all the groups with different doses of fangjihuangbai gelata. can "clearly be controlled and the number of leucocyte in mice can be reduced. Furthermore, the experimental results have the statistics significance.4. Fangjihuangbai gelata can reduce the content of PGE2 in inflammatory tissue,and the experimental results have the statistics significance.Conclusions1. Fangjihuangbai gelata has the analgesic effect by advancing the pain threshold and prolonging the ache delitescence in mice.2. Fangjihuangbai gelata has the anti-inflammatory effect by reducing the permeability of blood capillary, decreasing the phlogistic medium's effusion, controlling the phlogistic reaction and depressing the number of leucocyte in mice.3. Fangjihuangbai gelata can reduce the content of PGE2 in inflammatory tissue, which may be one of the mechanism of it can exert the analgesic and anti-inflammatory effect. |
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