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清胰汤对重症急性胰腺炎所致肠屏障损伤的治疗作用

Therapeutic effect of Qingyi decoction in severe acute pancreatitis-induced intestinal barrier injury.

作者信息

Zhang Jing-Wen, Zhang Gui-Xin, Chen Hai-Long, Liu Ge-Liang, Owusu Lawrence, Wang Yu-Xi, Wang Guan-Yu, Xu Cai-Ming

机构信息

Jing-Wen Zhang, Gui-Xin Zhang, Hai-Long Chen, Lawrence Owusu, Guan-Yu Wang, Cai-Ming Xu, Department of Surgery, The First Affiliated Hospital, Dalian Medical University, Dalian 116011, Liaoning Province, China.

出版信息

World J Gastroenterol. 2015 Mar 28;21(12):3537-46. doi: 10.3748/wjg.v21.i12.3537.

Abstract

AIM

To investigate the effect of Qingyi decoction on the expression of secreted phospholipase A2 (sPLA2) 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)-α 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 sPLA2 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 sPLA2 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-α and DAO 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 DAO and TNF-α, and the intestinal apoptosis index significantly correlated with sPLA2 expression in the intestine (r = 0.895, 0.893 and 0.926, respectively; Ps < 0.05). The levels of sPLA2, AMY, TNF-α, 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 sPLA2. This mechanism may be similar to that of verapamil.

摘要

目的

探讨清胰汤对肠道屏障损伤中分泌型磷脂酶A2(sPLA2)表达的影响。

方法

将50只健康的Sprague-Dawley大鼠随机分为对照组、重症急性胰腺炎(SAP)组、清胰汤治疗组(QYT)、地塞米松治疗组(DEX)和维拉帕米治疗组(VER)。通过向大鼠胆胰管逆行注入1.5%脱氧胆酸钠诱导建立SAP模型。在诱导SAP后24小时处死所有大鼠。采集每只大鼠的动脉血、肠道和胰腺进行检测。采用生化方法测定血清淀粉酶(AMY)和二胺氧化酶(DAO)水平,采用酶联免疫吸附测定法检测血清肿瘤坏死因子(TNF)-α水平。通过苏木精-伊红染色组织切片的显微镜检查观察采集组织的病理变化。分别采用逆转录聚合酶链反应和蛋白质印迹法检测sPLA2在mRNA和蛋白水平的表达。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道组织上皮细胞的凋亡情况。

结果

与对照组相比,SAP组sPLA2在mRNA和蛋白水平的表达均显著增加(分别为0.36±0.13 vs 0.90±0.38,以及0.16±0.05 vs 0.64±0.05;P<0.01)。血清中AMY、TNF-α和DAO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,以及10.37±2.44 U/L vs 37.89±5.86 U/L;P<0.01)。SAP大鼠和对照大鼠肠道上皮细胞的凋亡指数也有显著差异(0.05±0.02 vs 0.26±0.06;P<0.01)。血清DAO和TNF-α水平以及肠道凋亡指数与肠道中sPLA2表达显著相关(r分别为0.895、0.893和0.926;P<0.05)。与SAP组相比,QYT组、VER组和DEX组的sPLA2、AMY、TNF-α和DAO水平均降低,但与对照组相比无差异。然而,清胰汤干预对肠道屏障损伤的治疗效果最佳,尽管其治疗效果起效较慢。

结论

清胰汤通过抑制肠道sPLA2的过度表达改善急性胰腺炎诱导的肠道屏障损伤。该机制可能与维拉帕米相似。

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