柴芩承气汤对重症急性胰腺炎并肝损伤大鼠肝组织氧化应激的影响

Effect of Chaiqinchengqi decoction on hepatic tissue oxidative stress of rats with liver injury induced by severe acute pancreatitis

目的探讨柴芩承气汤对重症急性胰腺炎(SAP)并肝损伤大鼠肝组织氧化应激的影响。方法 72只Sprague Dawley大鼠随机分为假手术组、模型组和观察组,每组24只,各组再随机分为1、5、10 h 3个亚组,每组8只,假手术组大鼠打开腹腔后仅翻动胰腺组织数次,不给予任何干预;模型组和观察组大鼠采用去氧胆酸钠逆行胰胆管注射建立SAP大鼠模型。假手术组和模型组大鼠在造模成功后,1 h亚组在术后1 h,5 h亚组在术后1、5 h,10 h亚组在术后1、5、10 h,分别按10 m L·kg-1给予生理盐水灌胃1次;观察组1 h亚组大鼠在术后1 h,5 h亚组大鼠在术后1、5 h,10 h亚组大鼠在术后1、5、10 h,分别按10 m L·kg-1给予1 000 g·L-1柴芩承气汤灌胃1次。假手术组、模型组和观察组各亚组大鼠分别在末次灌胃后4 h麻醉处死大鼠,下腔静脉取血用于血清淀粉酶(AMS)检测;取胰腺组织和肝脏组织,用体积分数10%中性甲醛溶液固定,苏木精-伊红染色进行病理学观察;部分肝组织制备匀浆,进行超氧化物歧化酶(SOD)、丙二醛(MDA)、髓过氧化物酶(MPO)检测。结果假手术组1、5、10 h亚组大鼠胰腺组织结构清晰,腺泡小叶完整。模型组1 h亚组大鼠胰腺组织小叶间质水肿,可见大量炎性细胞浸润;5 h亚组大鼠胰腺组织水肿进一步增加,出血坏死、炎性细胞浸润明显;10 h亚组大鼠胰腺组织出现大面积腺泡细胞坏死。观察组1、5、10 h亚组大鼠胰腺损伤均明显减轻,仅见胰腺间质水肿和少量炎细胞浸润,坏死面积明显减少。假手术组1、5、10 h亚组大鼠肝细胞排列整齐,细胞质均匀,肝小叶及小叶中央静脉完整,轮廓清楚,肝细胞索网状纤维结构完整。模型组1、5 h亚组大鼠肝小叶界限不清,肝细胞索紊乱,肝窦变窄,肝细胞点状坏死和炎细胞浸润;10 h亚组大鼠可见大片肝细胞坏死。观察组1、5、10 h亚组大鼠肝细胞的水肿、炎症反应均有所减轻,肝小叶仅见少量炎症细胞浸润,未见明显坏死。与假手术组大鼠比较,模型组1、5、10 h亚组大鼠血清AMS水平均显著升高(P<0.05);与模型组大鼠比较,观察组1、5、10 h亚组大鼠血清AMS水平均显著下降(P<0.05);与假手术组大鼠比较,模型组1、5、10 h亚组大鼠肝组织SOD活性均显著下降(P<0.05),MDA含量、MPO活性均显著升高(P<0.05);与模型组大鼠比较,观察组1、5、10 h亚组大鼠肝组织SOD活性均显著升高(P<0.05),MDA含量、MPO活性均显著下降(P<0.05)。结论柴芩承气汤通过抑制氧化应激减轻胰腺、肝脏损害,对大鼠SAP合并肝脏损伤发挥治疗作用。

Objective To explore the effect of Chaiqinchengqi decoction on hepatic tissue oxidative stress of rats with liver injury induced by severe acute pancreatitis（ SAP）. Methods Seventy-two Sprague Dawley rats were randomly divided into sham operation group,model group and observation group,with 24 cases in each group. The rats in each group were randomly divided into 1,5,10 h three time points（ n = 8）. The rats in the sham operation group were only given abdomen opened and pancreatic tissue tipping without another intervention. The SAP rat model was established by deoxysodium cholate injected flyback through bile duct in model group and observation group. The rats in sham operation group and model group were given intragastric administration of physiological saline at the dose of 10 m L·kg- 11,5 and 10 h after the model was made successfully. The rats in observation group were given intragastric administration of 1 000 g·L- 1Chaiqinchengqidecoction treatment1,5 and 10 h after the model was made successfully. The rats were sacrificed 4 h after the last intragastric administration and blood from the inferior vena cava was collected to detecting serum amylase（ AMS）; the liver and pancreatic tissues was taken and treated with volume fraction of 10% neutral formalin fixed and stained with haematoxylin-eosin for pathological observation. Part of the liver tissue was made homogenate to detecting superoxide dismutase（ SOD）,malondialdehyde（ MDA） and myeloperoxidase（ MPO）. Results The pancreatic tissue structure of rats in 1,5 and 10 h subgroups of sham operation group was clear,and had complete acinus lobule. The pancreatic tissue structure of rats in model group 1 h subgroups showed interlobular edema and a large number of inflammatory cell infiltration,which in the rats of model group 5 h subgroups showed further increase edema,hemorrhage and necrosis,inflammatory cell infiltration. Large area of acinar cell necrosis was observed in the pancreatic tissue of the rats in the model group 10 h subgroups. The pancreatic injury in observation group 1,5 and 10 h subgroup rats were reduced significantly,only showed pancreatic edema and a small amount of inflammatory cell infiltration,necrosis area decreased significantly. In sham operation group,the rats in 1,5 and 10 h subgroups were arranged in order to arrange the liver cells,the cytoplasm was uniform,the hepatic lobule and central veins of the liver were complete,the contour was clear,and the structure of liver cell cords was intact. The pancreatic injury of rats in model group 1,5 h subgroups showed liver cell cord disorder,liver sinus narrowing,liver cell spott necrosis and inflammatory cell infiltration. The pancreatic injury of rats in model group 10 h subgroups showed large hepatic necrosis. The pancreatic injury of rats in observation group 1,5 and 10 h subgroups showed that liver cell edema,inflammatory reaction was diminished,the hepatic lobule only had a few inflammatory cell infiltration and had no obvious necrosis. Compared with sham operation group,the serum AMS level,liver tissue homogenate MDA,MPO level increased significantly while the liver tissue homogenate SOD level decreased significantly at each time point in model group（ P〈0. 05）. Compared with model group,the serum AMS level,liver tissue homogenate MDA and MPO were reduced significantly while the liver tissue homogenate SOD level was increased significantly at each time point in the observation group（ P〈0. 05）. Conclusion Chaiqinchengqi decoction soup can improve the liver tissue oxidative stress ability,and has a therapeutic effect on SAP complicated with liver damage.