Study On The Mechanism Of Yiqi Huayu Jiedu Prescription In Intervening ARDS Based On TLR4/NF-κB/NLRP3 Pathwa

ObjectiveTo use Yiqi Huayu Jiedu Decoction,an experienced prescription of Professor du Huitang of Dongzhimen Hospital of Beijing University of traditional Chinese Medicine,to interfere with acute lung injury/acute respiratory distress syndrome(ALI/ARDS)model rats induced by endotoxin(Lipopolysaccharide,LPS).Based on TLR4/NF-κB/NLRP3 signal pathway,the protective effects of Yiqi Huayu Jiedu Decoction on LPS-induced ALI rats and the levels of inflammatory mediators IL-1β,IL-18 and inflammatory protein molecules TLR4 and NLRP3 were studied through animal experiments.To further explore the mechanism of Yiqi Huayu Jiedu Decoction interfering with ARDS through TLR4/NF-κB/NLRP3 signal axis.Method1.Eighteen clean SD male rats were randomly divided into blank control group(n=6),model group(n=6)and traditional Chinese medicine group(n=6).After the end of adaptive feeding,the traditional Chinese medicine group was given Yiqi Huayu Jiedu Decoction diluted traditional Chinese medicine extract for 7 days,while the blank control group and model group were given distilled water for 7 days.After intragastric administration for 6 hours on the 7th day,the model group and traditional Chinese medicine group were made by tail vein injection of LPS(2mg/kg),and the blank control group was injected with 0.9%sodium chloride solution via tail vein.The animals were anesthetized by intraperitoneal injection of 3%pentobarbital sodium(dose 1ml/kg)16 hours after modeling,the abdominal aorta and lungs were exposed,abdominal aortic blood and right lung samples were collected for pathological observation,and lavage fluid was collected through left bronchoalveolar lavage.2.The samples of serum,bronchoalveolar lavage fluid(BALF),pathological sections of lung tissue stained with HE and lung homogenate were detected.The pathological changes of lung tissue and the smear count of BALF were observed under light microscope.The contents of IL-1β and IL-18 in serum and TLR4 and NLRP3 in BALF and lung homogenate were detected by enzyme linked immunosorbent assay(Elisa).Result1.Yiqi Huayu Jiedu Decoction can reduce the damage of lung tissue structure in rats with acute lung injury.Pathological observation showed that the lung tissue structure of rats in the blank control group was intact,there was no hyperemia and edema,and no inflammatory cell infiltration was found in the lung tissue.In the model group,the lung tissue structure was obviously damaged,the degree of intraalveolar hemorrhage,interstitial edema and alveolar wall collapse were significant,and a large number of inflammatory cells were infiltrated in the lung tissue.Compared with the model group,the traditional Chinese medicine group was relieved,the degree of bleeding and edema was improved,the exudation of inflammatory cells was reduced,and the destruction of lung tissue structure was alleviated.2.Yiqi Huayu Jiedu Decoction could reduce the contents of IL-1β,IL-18,TLR4 and NLRP3 in ARDS rats.The results of Elisa assay showed that the number of inflammatory factors such as IL-1β and IL-18 in serum of the model group was higher than that of the blank control group(P<0.001).The contents of TLR4 and NLRP3 inflammatory bodies in bronchoalveolar lavage fluid and lung homogenate were higher than those of the blank control group.The number of serum IL-1β and IL-18 inflammatory factors in the traditional Chinese medicine group was lower than that in the model group(P<0.001).The contents of TLR4 and NLRP3 inflammatory bodies in bronchoalveolar lavage fluid and lung homogenate were lower than those in the model group.Conclusion1.Yiqi Huayu Jiedu Decoction can reduce lung tissue injury and improve the degree of alveolar and pulmonary interstitial congestion,edema and inflammatory cell infiltration in rats with acute lung injury induced by endotoxin.2.The protective effect of Yiqi Huayu Jiedu Decoction on acute lung injury induced by lipopolysaccharide may be related to that Yiqi Huayu Jiedu Decoction can reduce the number of inflammatory factors such as IL-1β and IL-18 in the middle and lower reaches of TLR4/NF-κB/NLRP3 pathway in serum of ARDS rats.It can reduce the expression of TLR4 and NLRP3 inflammatory bodies in the upper reaches of TLR4/NF-κB/NLRP3 pathway in bronchoalveolar lavage fluid and lung homogenate of ARDS rats,inhibit the scorch death mediated by NLRP3 inflammatory bodies,and reduce the production of inflammatory mediators.