| Objective(1)To establish a rat model of natural moxibustion.(2)Observe the dynamic effect of natural moxibustion on the rat skin epidermis structure and inflammatory cell infiltration.(3)Observe the dynamic changes of blister and serum and leukocyte after natural moxibustion.Methods(1)In Experiment 1,cantharidin tincture solution was prepared with cantharidin powder(5% cantharidin tincture,10% cantharidin tincture,20% cantharidin tincture,30% cantharidin tincture).32 male SD rats were anesthetized and depilated,and 0.018 grams of cotton balls were filled in the center of PU waterproof blank paste plaster.male SD rats underwent anesthesia and depilation,filled 0.018 g cotton balls in the center of the PU blank waterproof plaster,then drew 200?l 5% cantharides tincture to soak cotton ball evenly and then applied it to stick and stimulate the skin,and added 200?l 5% cantharides tincture 2 hours later,then sealed.Collect the foam liquid in 4 hours.To Select the optimum foaming concentration of cantharidis tincture by observing the rate of foaming,the size of bubble,the volume of foam,the absorption of foam,the scab in foam area,the mortality rate and the number of total cells in bubble.(2)In experiment two,the concentration of cantharidin tincture was selected as the fixed concentration in Experiment 2.64 male SD rats were depilated with 4Cm2 in the neck,saline,and placed on a thermostatic table at room temperature of 26°C.Randomly divided into 8 groups: blank control group,95% alcohol group,natural moxibustion groups(12h,24 h,48h,72 h,96h),8 rats in each groups.(1)Rats in the blank group were given normal diet without intervention.(2)Rats in the 95% alcohol group were stimulated with an equal volume of 95% alcohol on the surface of the skin and then sealed.(3)the 4h group was foamed with 200?l 5% cantharidis tincture.After 4h,the blister was preserved.After anaesthetized by chloral hydrate,blood was taken from the Abdominal vein and the skin tissue was preserved for examination.(4)The remaining groups adopt the same treatment.Except for the blank group,rats in each group were housed in single cages.The dynamic changes of skin layer structure and inflammatory cell infiltration were observed by HE staining and optical microscope in all rats,the skin irritation was observed and scored,and the skin temperature was measured.ELISA was used to detect the content of TNF-? in serum and blister of rat.Blood routine test was used to observe the changes of white blood cells in the blood in rats.Results(1)200?l 5% cantharidis tincture is the best concentration of blistering,which had the best foaming effect,higher survival rate,more stable foam and less adverse reaction in rats.(2)After moxibustion,the skin epidermis of rats was thickened and swollen,slightly reddened,and blisters appeared.(3)The stimulation of cantharidin Tincture on the skin of rats is inflammatory stimulus,which has the characteristics of "red","swollen","hot" and "painful".(4)the content of TNF-alpha in the foaming solution was the highest in the 24 h group,suggesting that the inflammatory reaction was excessive in 24 h.Moreover,the inflammatory reaction appeared earlier and more significantly than that of blood.(5)The number of leukocytes in the blood and the different types of leukocytes are quite different.Conclusion(1)200?l 5% cantharidis tincture is the best concentration which can achieve stable blistering.(2)Natural Moxibustion is a kind of inflammatory stimulation,and it also has pain stimulating components.These two factors are synergistic and mutually exist.(3)For the blood,the inflammatory phase of the blisters appeared earlier and to a more significant degree,and the inflammatory response of the blisters was the most significant at 24 hours.The regulation of immune cells in the blood was mainly white blood cells,and the most significant increase was in 24 h. |
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