芍药苷治疗重症急性胰腺炎的实验研究

Effect of paeoniflorin treatment in rats with severe acute pancreatitis

目的观察芍药苷对重症急性胰腺炎(SAP)大鼠的治疗效果及对核因子kappaB(NF-κB)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)表达的影响。方法 SD大鼠40只随机分为5组:正常对照组(A组)、急性胰腺炎(AP)模型组24h点(B组)、AP模型组72h点(C组)、芍药苷治疗组24h(D组)和芍药苷治疗组72h(E组),每组8只。采用L-精氨酸腹腔内注射的方法诱发SAP模型,造模后立即灌胃生理盐水(B、C组)、芍药苷溶液(D、E组),分别于造模后24、72h时间点处死各组大鼠。采血检测血清淀粉酶、TNF-α、IL-6;取胰腺组织分2份,HE染色行光镜检查、WesternBlot法检测NF-κBp65蛋白表达。结果造模24h后与A组比较,B、D组大鼠血清淀粉酶均升高(P均<0.05),但D组淀粉酶水平明显低于B组(P<0.05);造模后72h,C、E组血清淀粉酶水平均下降至接近A组水平(P均>0.05)。D、E组较同时间点B、C组的病理改变和病理学评分均明显减轻(P均<0.05)。血清TNF-α和IL-6的水平分别为D组:(10.45±1.90)pg/ml和(610.38±106.19)pg/ml,E组:(9.93±1.63)pg/ml和(598.13±83.77)pg/ml;均明显低于同时间点的B组[(20.53±4.16)pg/ml,(1255.75±118.82)pg/ml,P均<0.05]与C组[(23.85±5.59)pg/ml,(1366.33±163.16)pg/ml,P均<0.05]。D、E组较同时间点B、C组NF-κBp65/β-actin的比值明显减少[D组与B组相比,(0.23±0.044)vs(0.71±0.029),P<0.05;E组与C组相比,(0.04±0.006)vs(0.65±0.026),P<0.05]。结论芍药苷对SAP大鼠治疗作用的机制可能与其抑制NF-κB活化,在整体水平上抑制细胞因子基因表达,使机体免受过量细胞因子的损伤等因素有关。

Objective To observe the therapeutic effect of paeoniflorin on severe acute pancreatitis （SAP） and the impact on the expressions of NF-κB ,TNF-α and IL-6 in rats. Methods Forty SD rats were randomly divided into five groups （n =8 each） ：normal control group （group A） ,AP 24 h model group （group B） ,AP 72 h model group （group C） ,paeoniflorin treatment 24 h group （ group D） and paeonifiorin 72 h treatment group （ group E）. The SAP model was induced by intraperitoneally injecting L-arginine. After the model was established, rats of groups B and C were treated with intra-gastric instillation of NS,and rats in groups D and E with intra-gastric instillation of paeoniflorin. The rats in group B, group D and group C,group E were respectively sacrificed at 24 or 72 hours after established model. The blood samples were taken for measureing serum amylase, TNF-α and IL-6. The pancreatic tissues were taken for analyzing pathological status of the pancreas by optical microscopy after HE staining and detecting the expression of NF-κB p65 protein in the pancreas by Western blot. Results Compared with A group,the serum amylase levels in groups B and D were obviously elevated 24 hours after estabilishing model （ all P 〈 0.05 ）, and the level in group D was lower than that in group B （ P 〈 0.05 ）. the levels in groups C and E were all declined to near A group＇s level 72 hours after estabilishing model （ all P 〉 0.05 ）. The pathological severity and pathological scores of the pancreas in group D and group E were more lighter than those in group B and group C at the same time point （ all P 〈 0.05 ）. Compared with groups B and C at the same time point, the levels of serum TNF-α and IL-6 were significantly decreased in groups D [ （ 10.45 ± 1.90） vs （20.53 ±4.16 ） pg/ml , P 〈 0.05 ; （ 610.38±106.19） vs （1255.75±118.82） pg/ml,P〈0.05]and group E[（9.93±1.63） vs （23.85±5.59） pg/ml,P〈0.05;（598.13±83.77） vs （1366.33±163.16） pg/ml,P 〈 0.05 ]. Compared with groups B and C at the same time point,the NF-κB P65/ β-actin ratio obviously decreased in group D [ （0.23±0. 044） vs （0.71±0. 029） ,P 〈0.05] and group E [ （0.04±0. 006） vs （0.65±0.026）, P 〈 0.05 ]. Conclusion Paeoniflorin may alleviate the pancreatic pathologic changes in SAP rats ,and its mechanism may be involved in inhibiting the activity of NF-κB and exnression of TNF-α and IL-6.