Kang Xin, Lu Xiao-Guang, Zhan Li-Bin, Liang Zheng-Kai, Guo Wen-Xiu, Ma Qi, Wang Yi, Song Jian-Bo, Feng Jin-Yu, Wang Cong-Han, Bai Li-Zhi, Song Yi, Liu Guo-Hui
Department of Emergency Medicine, Zhongshan Hospital, Dalian University, Dalian, 116001, China.
Basic Medical College, Nanjing University of Chinese medicine, Nanjing, 210023, China.
BMC Complement Altern Med. 2017 Jun 2;17(1):288. doi: 10.1186/s12906-017-1789-x.
Dai-Huang-Fu-Zi-Tang (DHFZT) is a famous traditional Chinese prescription with intestinal obstruction, acute pancreatitis and cholecystalgia for thousands of years. Our previous work found that DHFZT could act against pulmonary and intestinal pathological injury in rats with severe acute pancreatitis (SAP). But the underlying mechanism has not been fully elucidated. The aim of present study was to investigate whether DHFZT could relieve pulmonary and intestinal injury by regulating aquaporins after SAP induced by sodium taurocholate in rats.
Forty of SD rats were used for dose dependant experiments of DHFZT.Accurate-mass Time-of-flight liquid chromatography-mass spectrometry was used for qualitative screening of chemical compositions of DHFZT. Twenty-four rats were randomly divided into 3 groups: sham group (n = 8), model group (SAP, n = 8), DHFZT group (SAP with DHFZT treatment, n = 8). SAP models were established by retrograde injections of 5% sodium taurocholate solutions into rat pancreaticobiliary ducts. Blood samples were taken at 0, 12, 24, 48 h post-operation for detecting serum amylase, lipase, endotoxin, TNF-α, IL-6 and IL-10. Protein expression and location of aquaporin (AQP)1, 5, 8 and 9 were assessed by immunohistochemistry, western blot and immunofluorescence respectively.
The study showed that 27 kinds of chemical composition were identified, including 10 kinds in positive ion mode and 17 kinds in negative ion mode. The results showed that AQP1, AQP5 of lung, and AQP1, AQP5, AQP8 of intestine in model group were significantly lower than that of sham group (P < 0.05), and which were obviously reversed by treatment with DHFZT. In addition, protein levels of pro-inflammatory cytokines such as TNF-α, IL-6 and endotoxin in peripheral blood were significantly suppressed by DHFZT, and that anti-inflammatory cytokine like IL-10 was just opposite. Finally, we also noted that DHFZT reduced serum levels of amylase, lipase and endotoxin, and also improved edema and pathological scores of lung and intestine after SAP.
DHFZT ameliorated the pulmonary and intestinal edema and injury induced by SAP via the upregulation of different AQPs in lung and intestine, and suppressed TNF-α, IL-6 expression and enhanced IL-10 expression.
大黄芥子汤(DHFZT)是一种沿用数千年的著名中药方剂,用于治疗肠梗阻、急性胰腺炎和胆囊炎。我们之前的研究发现,DHFZT可对抗重症急性胰腺炎(SAP)大鼠的肺和肠道病理损伤。但其潜在机制尚未完全阐明。本研究旨在探讨DHFZT是否能通过调节水通道蛋白来减轻牛磺胆酸钠诱导的大鼠SAP后的肺和肠道损伤。
40只SD大鼠用于DHFZT的剂量依赖性实验。采用高分辨飞行时间液相色谱-质谱联用技术对DHFZT的化学成分进行定性筛选。将24只大鼠随机分为3组:假手术组(n = 8)、模型组(SAP,n = 8)、DHFZT组(SAP + DHFZT治疗,n = 8)。通过逆行注射5%牛磺胆酸钠溶液至大鼠胰胆管建立SAP模型。术后0、12、24、48 h采集血样,检测血清淀粉酶、脂肪酶、内毒素、TNF-α、IL-6和IL-10。分别采用免疫组织化学、蛋白质印迹法和免疫荧光法评估水通道蛋白(AQP)1、5、8和9的蛋白表达及定位。
研究表明,共鉴定出27种化学成分,其中正离子模式下10种,负离子模式下17种。结果显示,模型组肺组织的AQP1、AQP5以及肠组织的AQP1、AQP5、AQP8均显著低于假手术组(P < 0.05),而DHFZT治疗可明显逆转上述变化。此外,DHFZT显著抑制外周血中TNF-α、IL-6等促炎细胞因子及内毒素的蛋白水平,而抗炎细胞因子IL-10则相反。最后,我们还发现DHFZT降低了血清淀粉酶、脂肪酶和内毒素水平,并改善了SAP后肺和肠道的水肿及病理评分。
DHFZT通过上调肺和肠道中不同的AQP改善了SAP诱导的肺和肠道水肿及损伤,并抑制了TNF-α、IL-6的表达,增强了IL-10的表达。