| Objective: to investigate the protective effect and mechanism of chai huang qing yi huo xue granule on lung injury in rats with severe acute pancreatitis.Methods:(1)grouping: 128 healthy male SD rats were randomly divided into four groups according to the random number table: the blank group,the sham operation group,the model group,and the chai huang qing yi huo xue granules group(hereinafter referred to as chai huang group),with 32 rats in each group.(2)modeling: the rat model was established by injecting 5% sodium taurocholate 1ml/kg through duodenal papillary retrograde cholangiopancreatic duct.In the sham operation group,only the abdominal organs were turned over and the abdomen was closed.Blank groups do nothing.(3)preparation of drugs and drug delivery: The liquid extract of Chai Huang qing yi huo xue granules with 0.9% saline solution mixture of 0.12g/ml of liquids,the end of the building for each group of rats after waking,chai huang group to make a good solution to fill the stomach,calculated on the rat body weight,each 10 ml/kg,once every6 hours,lavage amount of normal saline control group,the blank group will not do.All rats were fasting and drinking water freely after the operation.(4)materials: 8 rats in each group were sacrificed at each time point of 6h,12 h,24h and 48 h after the operation.After laparotomy,the gross anatomical structure of the pancreas was observed,and blood was collected from the abdominal aorta.(5)detection: amylase and lipase were detected by automatic biochemical analyzer;The oxygen partial pressure and carbon dioxide partial pressure were measured by blood gas analyzer and the oxygenation index was calculated.The wet-dry weight ratio of lung tissue was calculated.The pancreas and lung tissues were stained with hematoxylin-eosin and scored by pathology.The concentrations of secreted phospholipase A2 and toll-like receptor 4 in lung tissues and secreted phospholipase A2 in intestinal tissues were determined by ELISA.The expressions of nuclear factor κB p65,tumor necrosis factor-α,interleukin-1β,intercellular adhesion molecule-1,aquaporin-1,JNK,and p38 MAPK in lung tissues were detected by immunohistochemistry.RT-PCR method was used to detect the expression of intercellular adhesion molecular-1,aquaporin-1,JNK1 and p38 MAPK m RNA in lung tissues.The expression of JNK,P38 MAPK and p-jnk,p-p38 MAPK in lung tissues were detected by western blot.(6)statistical methods: using SPSS17.0 software for statistical analysis of measurement data obey the normal distribution to represent,the comparison of different groups in each time point several times with repetitive measure analysis of variance,the comparison of different time points within each group and between groups of the same point in time is using single factor analysis of variance,between two groups to compare the LSD method.P <0.05 was considered statistically significant.Results:(1)compared with the blank group,control group of rats normally,pancreas and lung tissue no difference between the gross anatomy observation,the activity of amylase,lipase,lung wet dry weight ratio,pancreas and lung tissue pathological grading,partial pressure of arterial blood oxygen partial pressure,carbon dioxide,oxygen index level,the lung tissue of secretory phospholipase A2 type,concentration of toll-like receptor 4,nuclear factor kappa B p65 predominate,tumor necrosis factor-α and interleukin-1β,intercellular adhesion molecule-1,water channel protein-1,the expression of JNK,p38 MAPK,intestinal tissue concentrations of secretory phospholipase A2 type,The differences were not statistically significant(p>0.05).(2)compared with the sham operation group,the rats in the model group were generally in a poor condition after surgery,with such manifestations as vertical hair,lazy movement,lethargy,and decreased drinking water.After laparotomy,the pancreas was enlarged,with obvious hyperemia,edema,bleeding and necrosis.There were obvious white saponification spots on the pancreas and mesentery,and the pancreas was attached to the abdominal organs.After thoracotomy,a small amount of pleural effusion can be seen.The volume of the lung tissue becomes larger and turns dark red.There is congestion and edema.The hematoxylin-eosin staining of the pancreas showed structural disorder,inflammatory cell infiltration and blood cell exudation,and enlarged interlobular septa.Lung tissue hematoxylin-eosin staining showed extensive alveolar wall collapse and fusion,alveolar septum widened or collapsed,alveolar and interstitial edema and hemorrhage,fluid accumulation in alveoli showed atelectasis,a large number of white blood cells infiltrated and red blood cells exudated,and lung tissue structure was disordered.The activity of amylase and lipase increased.The pathological score of pancreas tissue,the wet/dry weight ratio of lung tissue and the pathological score were increased.The oxygen partial pressure and oxygenation index decreased and the carbon dioxide partial pressure increased.The expression levels of secreted phospholipase A2,toll-like receptor 4,nuclear factor κB p65,tumor necrosis factor-α,interleukin-1β,intercellular adhesion molecule-1,JNK and p38 MAPK proteins in lung tissues increased,and the expression levels of aquaporin-1proteins decreased.The concentration of secreted phospholipase A2 in intestinal tissues increased,and the differences were statistically significant(p<0.05).(3)compared with the model group,the general vital signs of the rats in the chaihuang group were improved,and the gross anatomy of the pancreas and lung tissues were observed.The activity of amylase and lipase decreased.The pathological score of pancreas tissue,the wet-dry weight ratio of lung tissue and the pulmonary pathology score were decreased.The oxygen partial pressure and oxygenation index increased while the carbon dioxide partial pressure decreased.The expression levels of secreted phospholipase A2,toll-like receptor 4,nuclear factor κB p65,tumor necrosis factor-α,interleukin-1β,intercellular adhesion molecule-1,JNK,and p38 MAPK decreased,and the expression levels of aquaporin-1 increased.The concentration of secreted phospholipase A2 in intestinal tissue decreased.The differences were statistically significant(p<0.05).Conclusion:(1)Chai huang qing yi huo xue granules can protect lung injury in rats with severe acute pancreatitis.(2)Chai Huang qing yi huo xue granules on severe acute pancreatitis model rats with lung injury protection mechanism may be related to the diversion channel on the expression of protein-1,cut nuclear factor κ B p65 predominate,tumor necrosis factor-α and interleukin-1β,intercellular adhesion molecule-1,secretory phospholipase A2,the expression of toll-like receptor 4,thereby blocking JNK/p38 MAPK signaling pathway. |
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