Therapeutic effect of Qingyi decoction in severe acute pancreatitis-induced intestinal barrier injury
Therapeutic effect of Qingyi decoction in severe acute pancreatitis-induced intestinal barrier injury
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DOI：
10.3748/wjg.v21.i12.3537
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发表时间：
2015-03-28
影响因子：
4.3
通讯作者：
Xu, Cai-Ming
中科院分区：
文献类型：
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作者：
Zhang, Jing-Wen;Zhang, Gui-Xin;Xu, Cai-Ming
目的：方法：健康SD大鼠50只，随机分为正常对照组、重症急性胰腺炎（SAP）组、清胰汤组、地塞米松组和维拉帕米组，每组10只。采用1.5%脱氧胆酸钠逆行胰胆管灌注法复制SAP模型。SAP诱导后24 h处死所有大鼠。采集每只大鼠的动脉血、肠和胰腺用于研究。采用生化方法测定血清淀粉酶（AMY）和二胺氧化酶（DAO）水平，采用酶联免疫吸附法测定血清肿瘤坏死因子（TNF）-α水平。通过苏木精和伊红染色组织切片的显微镜检查研究收获组织中的病理变化。逆转录聚合酶链反应（RT-PCR）检测sPLA（2）mRNA表达，Western blot检测sPLA（2）蛋白表达。采用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法检测肠上皮细胞凋亡情况。与对照组相比，SAP组5-PLA（2）mRNA和蛋白水平表达均显著增加（分别为0.36 +/- 0.13 vs 0.90 +/- 0.38和0.16 +/- 0.05 vs 0.64 +/- 0.05; Ps < 0.01）。血清中AMY、TNF-α和DA 0水平也显著升高（分别为917 62 U/L vs 6870 +/- 810 U/L、59.7 +/- 14.3 ng/L vs 180.5 +/- 20.1 ng/L和10.37 +/- 2.44 U/L vs 37.89 +/- 5.86 U/L; Ps < 0.01）。肠上皮细胞凋亡指数在SAP组和对照组之间也有显著性差异（0.05 ± 0.02vs0.26 ± 0.06; P < 0.01）。血清DA 0、TNF-α水平和肠细胞凋亡指数与肠组织5-PLA 2表达呈显著正相关（r分别为0.895、0.893和0.926; P < 0.05）。与SAP组相比，QYT、VER和DEX组的5 PLA（2）、AMY、TNF-α和DAO水平均降低，但与对照组相比无差异。然而，清胰汤干预，对肠屏障损伤的治疗效果最大，虽然其治疗效果的起效较慢。结论：清胰汤改善急性胰腺炎引起的肠屏障损伤，通过抑制肠道5 PLA 2的过度表达。这种机制可能与维拉帕米相似。
AIM: To investigate the effect of Qingyi decoction on the expression of secreted phospholipase A(2) (sPLA(2)) in intestinal barrier injury.METHODS: Fifty healthy Sprague-Dawley rats were randomly divided into control, severe acute pancreatitis (SAP), Qingyi decoction-treated (QYT), dexamethasone-treated (DEX), and verapamil-treated (VER) groups. The SAP model was induced by retrograde infusion of 1.5% sodium deoxycholate into the biliopancreatic duct of the rats. All rats were sacrificed 24 h post-SAP induction. Arterial blood, intestine, and pancreas from each rat were harvested for investigations. The levels of serum amylase (AMY) and diamine oxidase (DAO) were determined using biochemical methods, and serum tumor necrosis factor (TNF)-alpha level was measured by an enzyme linked immunosorbent assay. Pathologic changes in the harvested tissues were investigated by microscopic examination of hematoxylin and eosin-stained tissue sections. The expressions of sPLA(2) at mRNA and protein levels were detected by reverse transcriptase PCR and Western blot, respectively. A terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay was used to investigate apoptosis of epithelial cells in the intestinal tissues.RESULTS: Compared to the control group, the expression of 5PLA(2) at both the mRNA and protein levels increased significantly in the SAP group (0.36 +/- 0.13 vs 0.90 +/- 0.38, and 0.16 +/- 0.05 vs 0.64 +/- 0.05, respectively; Ps < 0.01). The levels of AMY, TNF-alpha and DA0 in serum were also significantly increased (917 62 U/L vs 6870 +/- 810 U/L, 59.7 +/- 14.3 ng/L vs 180.5 +/- 20.1 ng/L, and 10.37 +/- 2.44 U/L vs 37.89 +/- 5.86 U/L, respectively; Ps < 0.01). The apoptosis index of intestinal epithelial cells also differed significantly between the SAP and control rats (0.05 +/- 0.02 vs 0.26 +/- 0.06; P < 0.01). The serum levels of DA0 and TNF-alpha, and the intestinal apoptosis index significantly correlated with 5PLA2 expression in the intestine (r = 0.895, 0.893 and 0.926, respectively; Ps < 0.05). The levels of 5PLA(2), AMY, TNF-alpha, and DAO in the QYT, VER, and DEX groups were all decreased compared with the SAP group, but not the control group. Qingyi decoction intervention, however, gave the most therapeutic effect against intestinal barrier damage, although the onset of its therapeutic effect was slower.CONCLUSION: Qingyi decoction ameliorates acute pancreatitis-induced intestinal barrier injury by inhibiting the overexpression of intestinal 5PLA2. This mechanism may be similar to that of verapamil.