The Protective Effect Of Rhizoma Paridis Of Yunnnanensis On Rats With Acute Lung Injury Induced By Sepsis Through HMGB1-TLR4-NF-?B Signaling Pathway

Objective(s):To explore the effect of Rhizoma Paridis on inflammation regulation,and the expression of high mobility group box 1 and downstream TLR4-NF-?B signaling pathway in rats with acute lung injury induced by Cecal ligation and puncture(CLP).Methods:172 Sprague Dawley female rats,the clean grade,were randomly divided into 5 groups:sham group,ALI group,RP group,PDTC group,RP+PDTC group.CLP was used to establish the Septic-ALI model.The RP group was administrated with Rhizoma Paridis 20mg/kg in 2ml of saline twice a day 6 hours after modeling,and the PDTC group was injected 120mg/kg PDTC in 1ml of saline twice a day intraperitoneally 2 hours after the modeling.The sham group and the model group were administered intragastrilly and intraperitoneal with an equal amount of saline.The behavioral changes of rats were continuously observed,the survival time of rats in each group were recorded and a 7-day survival curve was drawn.Blood was taken from the abdominal aorta at 1d,3d,and 7d after treatment,and the left lung bronchoalveolar lavage operation was performed to obtain the lavage fluid.The automatic blood gas analysis was used to detect OI,Lac,and RI.ELISA was used to detect inflammatory factors(TNF-?,IL-6,IL-1?,HMGB1)in serum and bronchoalveolar lavage fluid,and lung injury biomarkers(sRAGE,SP-D,sICAM-1,CCSP)in bronchoalveolar lavage fluid;take 1d,7d lung tissue,weighed the gross lung tissue and calculated the wet/dry weight ratio to evaluate the degree of pulmonary edema;HE staining was used to evaluate the degree of lung tissue injury and inflammatory infiltration,TUNEL was used to evaluate lung tissue epithelial cell apoptosis in rats,immunohistochemical staining to detect the location of lung tissue HMGB1,TLR-4 and NF-?B p65 protein;qPCR and Western blot were used to detect HMGB1,TLR-4 and NF-?B p65 mRNA and protein expression levels in lung tissue.Results:?The Septic-ALI rat model was successfully produced.The 7-day survival rates of the 5 groups were 100%,35%,62.5%,50%,and 72.5%.Compared with the ALI group,the average survival time of the RP group and the RP+PDTC group was significantly improved(P<0.05).?Arterial blood gas analysis results showed that:compared with the Sham group,the OI of the ALI group was significantly reduced,and Lac?RI were significantly increased(P<0.05);compared with the ALI group,the 01 of the RP group?PDTC group began to increase significantly after 3 days of treatment and the RI and Lac were significantly decreased(P<0.05),the OI of the RP+PDTC group began to increase significantly after 1 day of treatment and the RI and Lac were significantly decreased(P<0.05).After combined treatment with RP and PDTC,blood gas analysis results were further improved compared with RP or PDTC single treatment.?The results of lung injury biomarkers showed that:compared with the Sham group,the sRAGE and sICAM-1 levels in the BALF of the ALI group increased significantly on 7 days(P<0.05),and SP-D and CCSP decreased significantly(P<0.05);compared with the ALI group,RP group?PDTC group?RP+PDTC group these lung injury biomarkers were significantly improved;RP and PDTC combined treatment,lung injury biomarkers levels were further improved than RP or PDTC single treatment.?The results of HE and TUNEL in lung tissue showed that compared with the sham group,the lung injury scores and the percentage of apoptotic cell in the ALI group increased significantly(P<0.05).Compared with the ALI group,the lung injury scores and the percentage of apoptotic cell in the RP group,PDTC group and the RP+PDTC group were significantly decreased(P<0.05).?Inflammatory factor results showed that:compared with the Sham group,TNF-?,IL-6,IL-1?,HMGB1 in the serum and BALF of the ALI group increased significantly at 1 day(P<0.05);compared with the ALI group,the RP group?PDTC group?RP+PDTC group,the levels of inflammatory factors were reduced,and the levels of inflammatory factors were further reduced as time goes on.In the RP+PDTC group,the levels of inflammatory factors were further reduced compared with RP or PDTC single treatment.?The results of qPCR,immunohistochemistry and western blot showed that compared with the sham group,the mRNA and protein levels of HMGB1,TLR4 and NF-?B p65 in the lung tissue of the ALI group were significantly increased(P<0.05),compared with the model group,HMGB1,TLR4 and NF-?B p65 mRNA and protein levels in the RP group,PDTC group and RP+PDTC group were decreased(P<0.05),compared to RP group,HMGB1,TLR4 and NF-?B p65 mRNA and protein levels in the RP+PDTC group were significantly decreased(P<0.05).Conclusion(s):?Rhizoma Paridis of yunnanensis inhibits the expression of inflammatory factors and HMGB1 in septic-ALI rats induced by cecal ligation and puncture,regulates the HMGB1-TLR4-NF-?B signaling pathway,and inhibits the inflammatory response,thereby reducing lung tissue damage and pulmonary edema,improving rat lung function and increasing survival rate.?The NF-?B inhibitor PDTC has similar inflammation regulation effect to Rhizoma Paridis of yunnanensis,and it has synergistic effect when combined with Rhizoma Paridis of yunnanensis.