| Objective In this study,SIRS model rats were made to explore the effects of scraping and bloodletting intervention on rectal temperature,white blood cell count and the content of inflammatory cytokines in SIRS model rats,as well as their protective effects on lung function.And explore whether the combination of different intervention order will have different effects.In order to provide clinical intervention strategies in the field of external treatment of body surface of traditional Chinese medicine.To provide new ideas and experimental basis for the treatment of systemic inflammatory response syndrome and other related infectious diseases.MethodsSixty-four SPF Wistar rats were randomly divided into 8 groups: blank group,model group,bloodletting group,scraping group,scraping + bloodletting group,bloodletting + scraping group,bloodletting with scraping group and ulinastatin group.All rats were measured rectal temperature before modeling.Then the rat models of systemic inflammatory response were prepared by intraperitoneal injection of 500 mg/kg yeast polysaccharide paraffin oil suspension during modeling,and the rats in the blank group were injected intraperitoneally with the same volume of sterilized liquid paraffin.The rats in each group were intervened at 2hours and 24 hours after modeling.Rats in Ulinastatin group were given intraperitoneal injection of Ulinastatin 27000 U/kg.Bloodletting group rats in 2h,24 h after modeling Shaoshang point bloodletting.Rats in the scraping group were scraped at Dazhui and Feishu points 2 h and 24 h after modeling.Bloodletting + scraping group was given Shaoshang point bloodletting 2 hours after modeling,Dazhui and Feishu points scraping 24 hours after modeling.Scraping + bloodletting group was scraped at Dazhui point and Feishu point 2hours after modeling,and bloodletting at Shaoshang point 24 hours after modeling.Bloodletting with scraping group were given Shaoshang point bloodletting and Dazhui and Feishu points scraping at 2h and 24 h after modeling.Rats in the blank group and the model group did not intervene,only simulated grasping action.The physical signs of the rats were observed and recorded,and the rectal temperature was measured at 2h,24 h,48h after modeling.And the peripheral blood leukocyte count was measured by the method of collecting blood from orbital venous plexus at 2 hours and 24 hours after modeling.Animals were collected within 48 h after modeling.The levels of inflammatory cytokines including TNF-α,IL-1β,IL-6,IL-10 and INF-γ in serum and inflammatory cytokines including TNF-α,IL-6,IL-10,INF-γ and TGF-β in lung tissue were detected by ELISA.The pathological changes of lung tissue and the score of lung injury were observed by HE pathological staining,and the average optical density of inflammatory cells was analyzed.Results1.Results of rectal temperature measurement: 2 hours after the establishment of the model,the rectal temperature of the rats in model group and intervention groups was lower than that in the blank group(P<0.05).At 24 hours after modeling,the rectal temperature in the model group was still lower than that in the blank group(P<0.05).The rectal temperature of rats in the bloodletting with scraping group and the bloodletting + scraping group was higher than that in the model group(P<0.05).And in the comparison among the intervention groups,the bloodletting with scraping group was significantly higher than the bloodletting group(P<0.05).At 48 hours after modeling,the rectal temperature in the model group was still significantly lower than that in the blank group(P<0.05).Compared with the model group,rectal temperature increased in bloodletting group,bloodletting + scraping group and scraping+ bloodletting group(P<0.05).2.Results of white blood cell count: at 2h after modeling,the white blood cell count of each group increased compared with the blank group,and then decreased significantly.At 24 hours after modeling,the white blood cell count in the model group was lower than that in the blank group(P<0.05).Compared with the model group,the white blood cell count increased in the bloodletting group and the bloodletting + scraping group(P<0.05).3.Pathological observation results of lung tissue: in the blank group,the alveolar structure was normal,clear and complete,and there were no obvious pathological changes.In the model group,there was obvious destruction of lung tissue structure,obvious thickening of alveolar wall,collapse,shrinkage and narrowing of alveolar cavity,inflammatory changes in pulmonary interstitium,and a large number of inflammatory cell infiltration in pulmonary interstitium and alveolar cavity.Some interstitial lung areas have red blood cell exudation,congestion,edema,pulmonary consolidation and other pathological changes.However,in the intervention group,there were still some alveolar fusion and alveolar wall thickening,but it had been significantly improved.Compared with the model group,the alveolar structure was clearer,and a small number of inflammatory cells were still seen in the alveolar cavity and pulmonary interstitium.4.Results of average optical density analysis of inflammatory cells in pathological sections of lung tissue: the average optical density of inflammatory cells in lung tissue in the model group was higher than that in the blank group(P>0.05).The average optical density of inflammatory cells in each group after treatment was significantly lower than that in the model group,and the difference was statistically significant(P<0.05).And the average optical density of inflammatory cells in the bloodletting + scraping group was significantly lower than that in the ulinastatin group and the bloodletting with scraping group(P<0.05).5.Results of lung tissue injury score: the score of lung injury in the model group was significantly higher than that in the blank group(P<0.05).Compared with the model group,it was significantly decreased in the bloodletting with scraping group,the bloodletting +scraping group and the scraping + bloodletting group(P<0.05).6.Results of enzyme-linked immunosorbent assay(ELISA)in rat serum: compared with the blank group,the serum levels of TNF-α,IL-1β,IL-6 and IFN-γ in the model group were significantly increased(P<0.05).Compared with the model group,the TNF-α in each intervention group was significantly lower than that in the model group(P<0.05).The serum IL-1β,IL-6 and IFN-γ levels in the bloodletting + scraping group were decreased,and the IL-10 level was increased(P<0.05).The content of IL-6 decreased and IL-10 increased in scraping + bloodletting group(P<0.05).The serum IL-6 and IFN-γ levels in the bloodletting group were decreased(P<0.05),the content of serum IL-6 in the bloodletting with scraping group was decreased(P<0.05).And the serum IL-6 content in the bloodletting + scraping group and the serum TNF-α and IL-6 content in the bloodletting with scraping group were lower than ulinastatin group and scraping group(P<0.05).7.Results of enzyme-linked immunosorbent assay(ELISA)in rat lung tissue: compared with the blank group,TNF-α,IL-6 and IFN-γ in the lung tissue of the model group were increased(P<0.05),and TGF-β content was decreased(P<0.05).Compared with the model group,the IFN-γ content in each intervention group was significantly decreased(P<0.05).TNF-αcontent in bloodletting group and bloodletting + scraping group decreased(P<0.05),the content of IL-6 in scraping group,bloodletting group,bloodletting + scraping group and scraping + bloodletting group decreased(P<0.05).The content of IL-10 in scraping group,bloodletting group,bloodletting + scraping group and scraping + bloodletting group increased(P<0.05).The content of TGF-β in bloodletting group was higher than that in model group and bloodletting scraping group(P<0.05).Conclusions1.Scraping and bloodletting therapy can improve the abnormal decrease of rectal temperature and white blood cell count in SIRS model rats.2.The combined use of bloodletting at Shaoshang acupoint and scraping at Dazhui and Feishu acupoint can reduce the acute inflammatory injury of lung tissue of SIRS model rats.Moreover,the levels of pro-inflammatory cytokines TNF-α,IL-1β,IL-6 and INF-γ in serum and lung tissue of SIRS model rats were decreased,and the levels of anti-inflammatory cytokines IL-10 and TGF-β were increased,so as to regulate the relative balance of pro-inflammatory and anti-inflammatory.3.The scraping and bloodletting therapy have different effects on the body through the intervention of different intervention order.In the intervention of SIRS model rats,the first bloodletting and then scraping group has a relatively better intervention effect than the intervention sequence combination of other groups. |
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