| Purpose:In this study,the E.coli endotoxin injection method was used to replicate the febrile model.The non-steroidal anti-inflammatory drug(indomethacin)was used as a control to observe the effects of the pricking and cupping therapy on the serum cytokine(IL-1β,GM-CSF,IL-6,TNF-α,IL-4,IL-10,IFN-γ)levels of endotoxin-induced rabbits,and protein expression levels of inducible nitric oxide synthase(i NOS),cyclooxygenase(COX-1 and COX-2)in hypothalamus tissue of rabbits.To reveal the mechanisms of the pricking and cupping method and partially explain its antipyretic advantage,providing experimental basis for mechanism explanation and clinical application of pricking and cupping therapy.Material and method: 1.Animals and groups: Twenty-five clean-grade rabbits were randomly divided into normal group,model group,pricking and cupping group,cupping group and western medicine treatment(indomethacin)group,with 5 rats in each group.2.Modelling method: Except for the normal rabbits,the other groups of rabbits used the E.coli endotoxin injection method to replicate the fever model,and the E.coli endotoxin was diluted with physiological saline 200 ng/ml,and the model was replicated by intravenous injection of rabbit ears at a dose of 200 ng/kg.3.Intervention approach: After the model was successfully replicated,the rabbits in the western medicine group were given an indomethacin suspension,and the intragastric dose was 10 ml /kg,model group rabbits,normal group rabbits,pricking and cupping group rabbits,and simply cupping group rabbits were given the same volume of normal saline for intragastric administration;the rabbits in simply cupping group and the pricking and cupping group were subjected to cupping operation 1 hour after successful modeling.The rabbits in the simple cupping group were fixed on the rabbit plate to expose the back.After shaving at the Dazhui point of the rabbit,a transparent plastic canister with an inner diameter of 25 mm was used to connect to the instrument.The canister pressure of -0.03 Mpa was used to extract the canister at Dazhui point in rabbits for 15 minutes;the rabbits in the pricking and cupping group were fixed and shaved as described above,first using a disposable needle(fixed penetration depth of 2 mm)to puncture the bleeding,and then cupping for 15 minutes at Dazhui;rabbits in normal group,model group,western medicine treatment(indomethacin)group were fixed in the same manner as rabbit plate,without cupping operation.4.Indicator Detection and Methods: The temperature was recorded once each hour after modeling to observe the changes in body temperature of the normal group,the model group,the pricking cupping group,the simple cupping group,and the western medicine treatment(indomethacin)group.The temperature was continuously measured for 8 hours,respectively.The rabbits in the normal group,the model group,the cupping group,the cupping group,in the western medicine group(indomethacin group)were subjected to blood collection from the ear vein.Enzyme-linked immunosorbent assay(ELISA)was used to detect the concentration of serum cytokines(IL-1β,GM-CSF,TNF-α,IL-6,IL-4,IL-10,IFN-γ)in the normal group,the model group,the pricking cupping group,the simple cupping group,and the western medicine treatment(indomethacin)group.After blood collection,the animals were sacrificed by air embolization(intravenous injection of 10 ml air),and the whole brain was quickly removed.The hypothalamus was taken between the optic chiasm and the gray nodules,and the specimen was placed in an EP tube and stored in liquid nitrogen.Immunoblotting(Western blot)was used to detect nitric oxide synthase(i NOS),cyclooxygenase(COX-1,COX-2)protein expression levels in the hypothalamic inducible type of rabbits in the normal group,the model group,the pricking and cupping group,the simple cupping group,and the western medicine treatment(indomethacin)group.5.Statistical methodology: All the data were represented by the averaged±standard deviation (?).The one-way analysis of variance was used for the comparison between the two groups,and the P<0.05 was considered statistically significant.Results: 1.After modeling,the temperature of the rectum gradually increased after modeling in the model group,and typical biphasic heat appeared.The body temperature rose to the peak at about 2h and 4h,which was significantly higher than that of the normal group.There was a statistically significant difference(P<0.01),which also suggested that the model was successfully replicated.After corresponding treatment,the temperature of rectum in pricking and cupping group,simple cupping group and Western medicine group rabbits was significantly lower than that in model group(P < 0.05),suggesting that the pricking and cupping therapy,simple cupping therapy and western medicine intervention had obvious antipyretic effects;there was no significant difference among the pricking and cupping group,the simple cupping group and the western medicine group(P>0.05),but from the numerical point of view,the rectal temperature of rabbits in the pricking and cupping group decreased more significantly than that in simple cupping group.The antipyretic effect of the pricking and cupping method was obviously superior to the simple cupping method.2.The serum levels of IL-1β,IL-6,GM-CSF and TNF-α in the rabbits of the model group were significantly higher than those in the normal group(P<0.01).After corresponding treatment,the levels of IL-1β,IL-6,GM-CSF and TNF-α in the serum of the pricking and cupping group,the simple cupping group and the western medicine group rabbits were significantly lower,and the pricking and cupping group and the western medicine group were significantly different from those in model group at the same time period(P<0.05).There were significant differences in serum levels of IL-1β,IL-6 and GM-CSF between the simple cupping group and the model group at the same time period(P < 0.05),but there was no significant difference compared with the level of TNF-a(P > 0.05).The results showed that the levels of IL-1β,IL-6,GM-CSF and TNF-α in serum cytokines of endotoxin-induced rabbits could be reduced by pricking and cupping,cupping and Western medicine intervention.There were no significant differences in the levels of serum IL-1β,IL-6 and GM-CSF between the three groups of rabbits in the cupping group,the simple cupping group and the western medicine group(P>0.05),while The levels of serum TNF-α in rabbits were statistically different in the simple cupping group and the other two groups(P<0.05).In general or numerically,compared with the simple cupping method,the pricking and cupping method can reduce the serum cytokine levels of the model rabbits more obviously,which is superior to the simple cupping therapy.3.The levels of serum IL-4 and IFN-γ in the model group were significantly lower than those in the normal group(P<0.05),and the serum IL-10 level was also decreased,but there was no statistical difference compared with the normal group.(P>0.05);After intervention,the levels of serum IL-4 and IFN-γ in the rabbits in the pricking and cupping group and the cupping group were significantly improved,which was statistically different from the model group(P<0.05).The levels of serum IL-10 in the two groups were still lower than those in the model group(P>0.05).The levels of serum IL-4,IFN-γ and IL-10 in the western medicine group were all after treatment.There was no significant improvement,and there was no significant difference between the two groups(P>0.05).There was no significant difference between the groups in the pricking and cupping group and the cupping group(P>0.05),but from the numerical point of view,the pricking and cupping group has a more advantageous role.It is suggested that the intervention of pricking and cupping method and simple cupping method can improve the levels of IL-4 and IFN-γ in serum cytokines of endotoxin-induced rabbits,but there is no significant effect on serum IL-10 level.Western medicine had no significant effect on serum IL-4,IFN-γ and IL-10 levels in rabbits.4.The expression of i NOS protein in the hypothalamus of rabbits in the model group was significantly higher than that in the normal group(P<0.05).After intervention,the expression of i NOS protein in the hypothalamus of the pricking and cupping group,the cupping group,the western medicine group was significantly lower than that in the model group.The decrease was statistically significant(P<0.05).There was no significant difference between the pricking and cupping group,the cupping group and the western medicine group(P>0.05).However,from the numerical point of view,the pricking and cupping therapy has a more dominant effect than the cupping therapy.5.After the model was established,the expression of COX-2 protein in the hypothalamus of rabbits in the model group was increased,which was statistically different from the normal group(P<0.05).However,the expression of COX-1 protein in the hypothalamus of the model group did not change significantly.There was no significant difference between the two groups(P>0.05).After intervention,the expression levels of COX-1 and COX-2 protein in the hypothalamus of the western medicine group were significantly lower than those in the model group(P<0.05);the expression of COX-2 protein in the hypothalamus of rabbits in the pricking and cupping group and the cupping group was significantly lower than that in the model group(P<0.05),but the expression level of COX-1 protein did not decrease significantly compared with model group(P > 0.05).There was no significant difference between the two groups in the pricking and cupping group and the cupping group;the expression levels of COX-1 and COX-2 protein in the rabbits were no significantly different(P>0.05),but from the numerical point of view,the pricking and cupping method has a more dominant effect than the simple cupping method.It indicated that the pricking and cupping method and simple cupping method had no inhibitory effect on the expression of COX-1 protein in the hypothalamic tissue of endotoxin-induced rabbits,and selectively inhibited the expression of central COX-2 protein.However,Western medicine(indomethacin)intervention significantly inhibited the expression levels of COX-1 and COX-2 proteins.Conclusion: 1.The pricking and cupping method has the same antipyretic effect as the western medicine treatment,and the pricking and cupping method has its advantage effect.2.In the process of antipyretic effect,the pricking and cupping method can reduce the levels of serum proinflammatory cytokines(IL-1β,GM-CSF,TNF-α,IL-6);it can improve some serum anti-inflammatory factors(IL-4,IFN-γ)levels and inhibit central i NOS and COX-2 protein expression levels,but have no inhibitory effect on central COX-1 protein expression levels.3.The Superiority mechanism of pricking and cupping method is that it can increase the level of serum anti-inflammatory factors(IL-4,IFN-γ)and inhibit central COX-2 protein.At the same time of expression level,there was no inhibition on the expression level of central COX-1 protein.4.The antipyretic effect of the pricking and cupping therapy is positive and safe to operate.It is worthy of clinical promotion. |
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