| Objective:In this study,the LPS/TLR4/NF-κB signaling pathway was used as the entry point to observe the inflammatory response of thermal moxibustion at Dazhui point on the airways of rats in an asthma model,which confirmed the good regulatory effect of thermal moxibustion on rats in an asthma model and provided an objective experimental basis for the prevention and heat sensitive of asthma by this method.Methods:After 50 SPF female rats were fed for 1 week,They were divided into blank group 8,model group 8,PS group 8,PDTC group 8,moxibustion group 18,and the asthma rat model was established by classical OVA sensitization.On the first and thirteenth days of the experiment,the model group was sensitized by intraperitoneal injection of 1 ml of 1%OVA solution containing 10 mg Al（OH）3.The blank group was injected with the same amount of normal saline intraperitoneally.Day 19 to 24:The model group was atomized with 8 ml 1%OVA solution for 30 min,while the blank group was atomized with the same amount of normal saline once a day.Day25-56:LPS group was intraperitoneally injected with 10 ug LPS 1 ml;In PDTC group,1 mg/kg PDTC 1 ml;was injected intraperitoneally;The other groups were injected with the same amount of normal saline intraperitoneally;In the moxibustion group,heat-sensitive suspension moxibustion was performed at Dazhui point for 40 min,and the skin temperature of the tail root of rats was recorded 5 min before moxibustion and every 5 min during moxibustion,and the temperature of the last 7 moxibustion sessions was counted.Combined with infrared thermal imaging,all the skin temperature fluctuations of the tail 1/3 were classified as heat-sensitive moxibustion group,and those below 1℃were classified as non-heat-sensitive moxibustion group,which was equal to 1℃.Finally,2 rats in the moxibustion group were excluded,including heat-sensitive moxibustion and non-moxibustion.From the 25th to the 56th day of the experiment,the intervention measures were once a day,three times a week,atomized by intraperitoneal injection and given 1 hour after suspension moxibustion with heat-sensitive moxibustion.The atomized liquid was the same as before in each group.During the experiment,the behavioral changes and mental state of rats were observed,the weight of rats before and after modeling was weighed,and the incubation period of asthma,the total duration of asthma attack and behavioral score were observed at the last atomization.The airway reactivity of rats was detected within 24 hours after the last atomization,and the samples were taken after the monitoring.Pathological changes of lung tissue in rats were observed by HE staining,expressions of TLR4,IL-4 and TNF-αin serum and BALF were detected by ELISA,expressions of IL-4,TNF-αand NF-κB m RNA in lung tissue were detected by Q-PCR,and expressions of TLR4 and NF-κB protein in lung tissue were detected by WB.Result:（1）general condition observationThe rats in the blank group showed no abnormal performance before and after modeling and were in good mental condition.In the model group,the rats in the PDTC,thermal moxibustion and non-thermal moxibustion groups were in good spirits,breathing was stable,and choking and coughing symptoms were significantly reduced,followed by the non-heat sensitive moxibustion group.（2）Body weight changeBefore modeling,there was no difference in body weight between the groups（P>0.05）.After treatment,the body mass of the model group and LPS group increased slowly,especially the weight of the LPS group was the lightest;the weight of the model group and LPS group decreased significantly compared with the rest of the intervention groups（P<0.05）;the weight of the PDTC group and the heat sensitive moxibustion group increased significantly compared with the non-heat sensitive moxibustion group（P<0.05）.（3）Wheezing latency,total duration of asthma attack and behavioral scoreCompared with the blank group,the latency of asthma induction in each intervention group was significantly reduced,and the total duration of asthma attack was increased,with statistical significance（P<0.05）.Compared with the model group,the latent period of asthma,the total duration of asthma attack and the behavioral score of PDTC group,heat-sensitive moxibustion group and non-heat-sensitive moxibustion group were different（P<0.05）.And the difference between heat-sensitive moxibustion group and non-heat-sensitive moxibustion group was statistically significant（P<0.05）.（4）Effect of heat-sensitive moxibustion on airway reactivity（Penh）in ratsCompared with the blank group,the Penh value of each intervention group was significantly decreased（P<0.05）.Penh value in LPS group was significantly higher than that in model group（P<0.05）.Compared with the model group,the Penh values of all groups except the blank group decreased significantly（P<0.05）.Penh values in PDTC group and heat-sensitive moxibustion group were significantly lower than those in non-heat-sensitive moxibustion group（P<0.05）.（5）HE staining results of rat lung tissueThe infiltration of inflammatory cells in lung tissue of rats in blank group was less.In the model group,the wall of the rat was thickened and narrowed,and a large number of inflammatory cells infiltrated in the cavity.In LPS group,the lumen of rats was severely deformed and inflammatory cells infiltrated seriously.The airway epithelial tissue of rats in PDTC group and heat-sensitive moxibustion group was basically intact,and inflammatory cells infiltrated less.The improvement of lung tissue morphology in non-heat-sensitive moxibustion group was second to that in heat-sensitive moxibustion group.（6）Effect of heat sensitive moxibustion on the expression of TLR4,IL-4 and TNF-αin rat serumCompared with the blank group,the expressions of serum TLR4,IL-4 and TNF-αin the model group were significantly increased（P<0.05）,while the expressions of serum TLR4,IL-4 and TNF-αin PDTC and heat-sensitive moxibustion group were significantly decreased compared with those in non-heat-sensitive moxibustion group（P<0.05）.（7）Effect of thermosensitive moxibustion on the expression of TLR4,IL-4 and TNF-αin rat BALFCompared with the blank group,TNF-αexpression in BALF was significantly higher in all intervention groups（P<0.05）,TLR4 and IL-4 in BALF of LPS group,model group and non-heat-sensitive moxibustion group were also significantly increased（P<0.05）;TLR4,IL-4 and TNF-αcontents in BALF were significantly increased in the model and LPS groups compared with all groups except the blank group（P<0.05）;TLR4,IL-4,and TNF-αexpression in BALF of PDTC and heat sensitive moxibustion groups were significantly decreased compared with non-heat sensitive moxibustion groups（P<0.05）.（8）Effect of heat sensitive moxibustion on IL-4,TNF-α,and NF-κB m RNA expression in lung tissue.Compared with the model group and LPS group,the expression of IL-4,TNF-α,and NF-κB m RNA was significantly lower in the PDTC,heat sensitive moxibustion,and non-heat sensitive moxibustion groups（P<0.05）;and the expression of IL-4,TNF-α,and NF-κB m RNA was significantly increased in the LPS group compared with the model group（P<0.05）;the expression of IL-4,TNF-α,and NF-κB m RNA expression was significantly lower in the PDTC and heat sensitive moxibustion groups compared with the non-heat sensitive moxibustion group（P<0.05）.（9）Effect of thermosensitive moxibustion on TLR4,NF-κB protein expression in rat lung tissuesTLR4 and NF-κB protein expression was significantly lower in the remaining groups compared to the model and LPS groups（P<0.05）;TLR4 and NF-κB protein expression was significantly lower in the PDTC and heat sensitive moxibustion groups compared with the non-heat sensitive moxibustion group（P<0.05）.Conclusion:（1）Thermal moxibustion can significantly improve the body weight of asthmatic rats,prolong the incubation period of asthma induction,reduce the duration of asthma attack,improve the lung function of rats,reduce the airway response and inflammatory cell infiltration in lung tissue,indicating that heat sensitive moxibustion has a good regulatory effect on the airway inflammatory response of asthmatic rats.（2）Heat sensitive moxibustion can exert therapeutic effects on asthma by reducing LPS,TLR4,and NF-κB levels,inhibiting LPS/TLR4/NF-κB signaling pathway activation,and suppressing airway inflammatory responses. |
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