| Objective: In this study,a rat model of obstructive jaundice was prepared by common bile duct ligation.Our aim is to explore the role of SIRT1-NLRP3 signaling pathway in the pathogenesis of lung injury induced by obstructive jaundice by intraperitoneal injection of SIRT1 inhibitor(EX527).At the same time,the therapeutic effect of Yinchenhao Decoction was observed by intragastric administration.We hope to provide relevant theoretical basis and experimental basis for the treatment of obstructive jaundice lung injury by integrated Chinese and Western medicine.Methods: Forty SPF male Sprague-Dawley rats were randomly divided into four groups with 10 rats in each group: Control group(CON group),SIRT1 inhibitor Selisistat intervention group(EX527 group),Bile duct ligation(BDL group)and Yinchenhao Decoction intervention group(YCHD group).After 7 days of adaptive feeding,common bile duct ligation was performed,12 h of fasting was performed before surgery,skin preparation and disinfection were performed after anesthesia,and a small incision of about 2cm was made under xiphoid process along the midline of the abdomen.After exposure,the abdomen was dissociated,common bile duct ligation was performed,and the abdomen was closed for suture.EX527 group was intraperitoneally injected with 5mg/ml/2d EX527 solution after operation.YCHT group was given2g/0.25kg/d Yinchenhao Decoction by intragastric administration.CON group and BDL group were given the normal saline with equal dose at the same time.The rats in each group were anesthetized 14 days after modeling,we collected their serum,liver tissue,lung tissue and alveolar lavage fluids.The right upper lung was immediately fixed with4% paraformaldehyde solution after sampling.The degree of pathological injury of liver and lung tissue in each group was evaluated by histological sections.Alveolar lavage was performed on the right middle lung.And the wet/dry weight ratio of the lower right lung was determined(W/D).We determined Lung tissue myeloperoxidase activity(MPO),protein concentration in alveolar lavage fluid and serum liver function indexes:total bilirubin(TBIL),direct bilirubin(DBIL),alanine aminotransferase(ALT)and aspartate aminotransferase(AST).ELISA was used to determine the pro-inflammatory factors in serum and alveolar lavage fluid.The m RNA and protein expression situation in lung tissues of SIRT1,NLRP3 and NF-k B was tested by RT-PCR,western blotting and immunofluoresence.Results: Compared with CON group,liver cord structure was disordered,liver tissue hyperplasia was accompanied by inflammatory cell infiltration,lung tissue hemorrhage and edema was accompanied by inflammatory cell infiltration,liver and lung tissue pathological scores were increased(p < 0.001),lung tissue W/D was significantly increased(p < 0.05),lung tissue myeloperoxidase activity was increased(p < 0.001).Protein concentration in alveolar lavage fluid was increased(p < 0.001),liver function indexes in serum were significantly increased(p < 0.001),in serum and alveolar lavage fluid,the pro-inflammatory factor IL-6 and TNF-α contents were significantly increased(p < 0.05).SIRT1 m RNA and protein expression were decreased(p < 0.01),and NLRP3 and NF-k B m RNA content were increased(p < 0.01).SIRT1 gene and protein expression levels were significantly is inhibited in EX527 group(p < 0.01),but the NLRP3 gene expression levels were increased(p < 0.05),pro-inflammatory factor release in serum and alveolar lavage fluid was increased,and the pathological damage of liver and lung tissue is more serious than BDL group(p < 0.05).The results indicated that inhibition of SIRT1 might aggravate lung inflammation by activating NLRP3.The pathological injury of liver and lung tissue in YCHD group was less than that in BDL group,and the W/D of lung tissue,MPO activity,protein contents in alveolar lavage fluid,liver function indexes and pro-inflammatory factors in serum and alveolar lavage fluid were significantly reduced(p < 0.05),suggesting that Yinchenao decoction could alleviate lung injury caused by obstructive jaundice in rats.In contrast to the results of EX527 group,SIRT1 gene and protein in YCHD group were high expressed,while the NLRP3 was low expressed,suggesting that Yinchenhao decoction may inhibit the activation of NLRP3 through up-regulation of SIRT1,thus alleviating lung injury caused by obstructive jaundice.Conclusions: 1.The rat model of obstructive jaundice can cause the lung injury.Liver tissue and lung tissue pathological injury was obvious,and the injury index in serum was significantly increased can proved it,the W/D ratio of lung tissue was significantly rise,and the concentration of alveolar perfusion fluid was increased,which indicated that the lung tissue of the rats with obstructive jaundice was damaged in different degrees of structure and function,and the rat model of obstructive jaundice lung injury was successfully prepared.2.Obstructive jaundice lung injury was negatively correlated with SIRT1 expression water and positively correlated with NLRP3 expression.In EX527 group,inhibition of SIRT1 expression promoted NLRP3 expression and aggravated lung inflammation injury.This suggests that the mechanism of lung injury induced by obstructive jaundice may be to promote the release and activation of NLRP3 by down-regulating the expression of SIRT1,thus producing a large number of inflammatory factors to induce the activation of inflammatory cells and cause lung injury.3.Yinchenhao Decoction can inhibit the activity of NLRP3 by promote the activity of SIRT1,reduce the concentration of pro-inflammatory factors in lung,and alleviate lung injury caused by obstructive jaundice. |
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