Experimental Study of Tongchangqingyi Decoction Regulating TLR4/NF-κB Pathway to Reduce Lung and Intestinal Injury in Rats with Severe Acute Pancreatitis

ZHANG Qian; LIU Binbin; LI Siqi; SU Chengxia; HUANG Chaoqun; HU Yunlian

doi:10.13730/j.issn.2097-2148.2024.10.001

Abstract：Objective To investigate the protective effects of Tongchangqingyi Decoction on lung and intestinal injury in rats with severe acute pancreatitis(SAP)and its possible mechanisms.Methods 24 SPF male SD rats were randomly divided into sham operation group,model group,Tongchangqingyi Decoction group and ulinastatin group,with 6 rats in each group.After the rats of sham operation group were anesthetized and exposed to the abdominal cavity,the duodenum and pancreas were gently turned several times and the abdomen was sutured,SAP rat models were prepared by retrograde injection of sodium taurine cholate through bile pancreatic duct puncture in other groups.The rats were given the drug at 4 hours after the modeling was successful.Rats of Tongchangqingyi Decoction group were given Tongchangqingyi Decoction by intragastric administration,rats in model group and sham operation group were given equal volume normal saline instillation,ulinastatin group rats were given uli-nastatin injection by tail vein,once every 8 hours,for a total of 3 times.24 hours after the modeling,the rats were anesthe-tized and sampled,the contents of amylase(AMS)and lipase(LPS)in serum were determined by biochemical method,se-rum levels of C-reactive protein(CRP),tumor necrosis factor alpha(TNF-α),interleukin 6(IL-6)and interleukin 10(IL-10)were detected by enzyme-linked immunosorbent assay(ELISA),the pathological changes of pancreatic,colon and lung tissues of rats were observed by hematoxylin-eosin(HE),and the contents of occludin,claudin and zonula occludens-1(ZO-1),protein expression of Toll-like receptor 4(TLR4)and nuclear transcription factor kappa B(NF-κB)in colon tis-sue and lung tissue of rats were detected by Western blot.Results HE staining showed no significant pathological changes in pancreas,colon and lung tissues in the sham operation group.In the model group,pancreatic acinous space was signifi-cantly expanded,interstitial edema,lobular space was enlarged,a large number of inflammatory cells infiltrated and tissue sheet necrosis was observed,colon cells were swollen,inflammatory cells were infiltrated,mucosal glands were reduced and intestinal villi was exfoliated.Lung tissue cell edema,inflammatory cell infiltration and interstitial bleeding were obvious in the model group.Compared with the model group,The destruction of pancreatic acinar structure,interstitial edema,hem-orrhage and necrosis were alleviated in Tongchangqingyi Decoction group and ulinastatin group,the inflammatory cell infil-tration of colon was reduced,the pulmonary tissue cell edema and inflammatory infiltration were also alleviated.Compared with sham operation group,the contents of serum AMS,LPS,CRP,TNF-α and IL-6 in model group were significantly increased(all P＜0.01),the content of IL-10 was significantly decreased(P＜0.01),the protein expressions of Occludin,Claudin and ZO-1 in colon tissue were significantly decreased(all P＜0.01);compared with model group,the contents of AMS,LPS,CRP,TNF-α and IL-6 in serum of Tongchangqingyi Decoction group and ulinastatin group were significantly decreased(all P＜0.01),and the content of IL-10 was significantly increased(P＜0.01),the expression of Occludin,Claudin and ZO-1 in colon tissue were significantly increased(all P＜0.01),while the protein expressions of TLR4 and NF-κB p65 in colon tissue were significantly decreased(all P＜0.01).Compared with ulinastatin group,the contents of serum AMS,LPS and CRP in Tongchangqingyi Decoction group were significantly increased(all P＜0.05),the content of IL-10 was significantly decreased(P＜0.01),and the protein expression of Occludin and ZO-1 in colon tissue was sig-nificantly decreased(all P＜0.05),the protein expression levels of TLR4 in colon and lung were significantly increased(all P＜0.05),but the contents of TNF-α and IL-6 in serum,the expression levels of Claudin and NF-κB p65 in colon and NF-κB p65 in lung were not statistically significant(all P＞0.05).Conclusion Tongchangqingyi Decoction can alleviate lung-intestinal injury,reduce the release of inflammatory factors and protect intestinal mucosal barrier in rats with severe acute pancreatitis,and its mechanism may be related to the regulation of TLR4/NF-κB pathway.