【作者】 石星亮；

【导师】 陈垦；

【作者基本信息】 广东药学院 ， 消化内科， 2010， 硕士

【摘要】 目的应用中药新药清胰颗粒(qingyi grains, QYG)对重症急性胰腺炎(severeacute pancreatitis，SAP)大鼠进行治疗，观察模型动物血清淀粉酶变化，对胰腺、肺、肠、肝、肾等组织进行病理组织学检查，观察其治疗效果；观察模型动物血清过氧化物岐化酶（superoxide dismutase, SOD）和丙二醛（malondialdenhyde, MDA）等氧化应激因子变化；观察模型动物各器官核因子κB（NF-κB）和高迁移率族蛋白1（HMGB1）的变化，探讨中药新药清胰颗粒的可能的治疗机制，为日后投入临床使用提供实验依据。方法根据随机对照试验将Sprague-Dawley大鼠分为：假手术组(sham-operation group, sham)，SAP组，QYG组，每组18只，各组又分别随机分为24h、48h、72h三组，每组6只。采用逆行胰胆管内注射5%牛磺胆酸钠(sodiumtaurocholate，STC)（0.1ml/100g)制备大鼠SAP模型。QYG组于造模后立即给予清胰颗粒溶液（1ml/100g）灌胃治疗，之后每12h重复给药一次，sham组仅予开腹后翻动胰腺，不予胰胆管注射。各组动物于相应时间点处死收集血液和各器官组织，人工碘比色法测定血清淀粉酶，羟胺比色法测定血清总SOD含量，硫代巴比妥酸（TBA）比色法检测血清MDA含量，HE染色观察胰腺、肺、小肠、肝、肾等组织病理，S-P免疫组化法观察胰腺、肺、小肠、肝、肾等组织NF-κBp65以及HMGB1表达变化。结果1血清淀粉酶变化SAP组与QYG组血清淀粉酶水平明显高于sham组；与SAP组比较，QYG组于24h和48h血清淀粉酶明显降低（t=7.929，P=0.000；t=9.847，P=0.000），而与QYG治疗后72h无统计学差异(t =0.769，P=0.460)。2血清SOD和MDA含量变化SAP组与QYG组各时间点的血清总SOD含量均明显低于sham组。于QYG治疗后48h和72h，QYG组血清SOD较SAP组明显升高（t =-5.483，P=0.000；t=-4.519，P=0.001），而于治疗后24h无统计学差异(t =-0.747，P=0.472)。SAP组与QYG组各时间点的血清MDA含量均明显高于sham组。于QYG治疗后24h、48h和72h，血清MDA含量均较SAP组明显降低（t =7.128，P=0.000；t =4.544，P=0.001；t =5.501，P=0.001）。3各器官的病理组织学变化SAP大鼠胰腺、肺、小肠、肝、肾等组织出现了不同程度的损伤。应用QYG治疗后的大鼠各器官的病理组织学损伤程度较SAP组有所改善。4各器官NF-κBp65、HMGB1表达变化SAP大鼠胰腺、肺、小肠、肝、肾等组织NF-κBp65、HMGB1表达增加，应用QYG治疗可不同程度的减少NF-κBp65、HMGB1表达。结论1 SAP大鼠血清淀粉酶明显升高，清胰颗粒可以降低急性胰腺炎大鼠高血清淀粉酶血症。2 SAP大鼠胰腺、肺、小肠、肝、肾等器官出现了不同程度的损伤。应用清胰颗粒治疗后的大鼠，各器官的损伤程度较SAP组改善。中药新药清胰颗粒对急性胰腺炎大鼠的多器官损伤有保护作用。3 SAP时大鼠机体抗氧化能力减弱，产生大量氧自由基，应用清胰颗粒治疗可提高机体清除氧自由基的能力，降低氧化应激因子的水平。抗氧化作用可能是清胰颗粒治疗急性胰腺炎的可能机制之一。4 SAP大鼠胰腺、肺、小肠、肝、肾等器官NF-κBp65、HMGB1表达增加，应用清胰颗粒治疗后，大鼠各器官NF-κBp65、HMGB1表达不同程度的减少。清胰颗粒可通过抑制各器官NF-κBp65、HMGB1表达，从而发挥其器官保护作用。更多还原

【Abstract】 Objective severe acute pancreatitis (SAP) rats were treated by the Chinese newmedicine Qing Yi grains (QYG). To observe the treatment effect, changes of serumamylase in animal models were observed. Pancreas, lung, intestine, liver, kidney wereharvested for pathological examination. Changes of serum peroxide dismutase (SOD)and malondialdehyde (MDA) were observed. Expression changes of nuclear factorκB (NF-κB) and high mobility group protein 1 (HMGB1) in different organs wereobserved to explore the possible treatment mechanism of Chinese new medicine QYGand provide experimental basis for clinical use.Methods Sprague-Dawley rats were randomly divided into sham operation group,SAP group and QYG group. Each group contained 18 rats and then randomly dividedinto 24h, 48h, 72h three groups, n=6. Rats were prepared by retrograde pancreaticduct injection of 5% sodium taurocholate (STC) (0.1ml/100g) to establish SAP model.QYG group received Qing Yi grains solution (1ml/100g) gavage treatmentimmediately after operation. Repeated administration was given every 12h. Animalsin each group were sacrificed at corresponding time points to collect the blood andorgans to detect serum amylase, T-SOD, MDA, HE staining for pancreas, lung,intestine, liver and kidney, S-P immunochemistry for expressions of organs NF-κBp65and HMGB1. Results1 Changes of the serum amylaseSerum amylase levels in SAP group and QYG group were significantly higherthan sham operation group. Compared with SAP group, serum amylase levels in QYG24h and 48h group were significantly decrease (（t =7.929，P=0.000；t =9.847，P=0.000）). They were no significant difference in 72h (t =0.769，P=0.460)2 Changes of serum SOD and MDASerum SOD levels in SAP group and QYG group were significantly lower thanthe sham operation group. At 48h and 72h after QYG treatment, serum SOD levelswere significantly higher than SAP group (t =-5.483，P=0.000；t =-4.519，P=0.001).There were no significant difference at 24h group (t =-0.747，P=0.472). Serum MDAin SAP group and QYG group were significantly higher than sham operation group ateach time point. Compared with SAP group, serum MDA levels of QYG group weresignificantly lower at each time point (t =7.128，P=0.000；t =4.544，P=0.001；t=5.501，P=0.001).3 Pathological changes in organsPathological changes occured in SAP rats’pancreas, lung, intestine, liver, kidney.QYG treatment can improve pathological changes in SAP rats’organs.4 Changes of NF-κB, HMGB1 expression in organsNF-κB, HMGB1 expression increased in SAP rats’pancreas, lung, intestine,liver, kidney. Organs NF-κB, HMGB1 expression decreased to different extent afterapplication of QYG treatment.ConclusioConclusion1 Serum amylase in SAP rats significantly increases. QYG can reduce high serumhyperamylasemia in SAP rats.2 In SAP rats, pancreas, lung, small intestine, liver and kidney showed damge invarying degrees. After application of QYG treatment, the extent of organ damageimproves. Traditional Chinese drug Qingyi grains can protect multiple organ damagein rats with acute pancreatitis. 3 In SAP rats, antioxidant capacity is decreased, resulting in a large number of oxygenfree radicals. QYG can increase the antioxidant capacity of body and reduce the levelof oxidative stress factors. Antioxidant effect may be one of the therapy mechanism ofQYG.4 NF-κB, HMGB1 expression increased in SAP rats’pancreas, lung, intestine, liver,kidney. Organs NF-κB, HMGB1 expression decreased to different extent afterapplication of QYG treatment. QYG can inhibit the organ NF-κB, HMGB1expression, and thus play its role in organ protection.更多还原