«上一篇/Previous Article|本期目录/Table of Contents|下一篇/Next Article»

《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

30卷

期数:

2024年05期

页码:

415-421

栏目:

基础医学

出版日期:

2024-09-25

文章信息/Info

Title:

Study on the mechanism of Xuebijing injection in treating severe acute pancreatitis with lung injury

文章编号:

1006-8147（2024）05-0415-07

作者:

王舜¹; 张桂贤²; 冯志乔³; 沈洪昇²; 李文畅²; 宗文辉²; 蔡隽²; 刘洪斌²

（1.天津医科大学研究生院，天津 300070；2.天津市医药科学研究所，天津 300020；3.天津红日药业股份有限公司，天津 301700）

Author(s):

WANG Shun¹; ZHANG Guixian²; FENG Zhiqiao³; SHEN Hongsheng²; LI Wenchang²; ZONG Wenhui²; CAI Jun²; LIU Hongbin²

（1.Graduate School of Tianjin Medical University，Tianjin 300070，China；2.Tianjin Institute of Medical & Pharmaceutical Sciences，Tianjin 300020，China；3.Tianjin Chase Sun Pharmaceutical Co.，Ltd. Tianjin 301700，China）

关键词:

重症急性胰腺炎; 急性肺损伤; 血必净注射液

Keywords:

severeacute pancreatitis; acute lung injury; Xuebijing injection

分类号:

R285.5

DOI:

10.20135/j.issn.1006-8147.2024.05.0415

文献标志码:

A

摘要:

目的：探讨血必净（XBJ）注射液对重症急性胰腺炎（SAP）大鼠肺损伤模型的治疗机制。方法：将80只雄性Sprauge-Dawley（SD）大鼠随机分为4组：对照组、SAP模型组、XBJ低剂量治疗组、XBJ高剂量治疗组，每组20只。除对照组，各组大鼠均经胰胆管匀速逆行注射4.5%牛磺胆酸钠溶液（0.1 mL/100 g，0.05 mL/min）以诱发SAP，造模成功30 min后，将XBJ低、高剂量治疗组大鼠经尾静脉注射XBJ（剂量分别为5、20 mL/kg），对照组及模型组给予等体积生理盐水。24 h后，从每组中随机抽取10只大鼠，经尾静脉注射Evans blue检测肺组织毛细血管通透性。处死余下的各组大鼠，取腹水测量其体积；取部分胰腺及肺组织计算组织干湿重比值；苏木精-伊红（HE）染色观察胰腺、肺脏的病理改变，并进行病理评分；酶联免疫吸附法（ELISA）检测腹水淀粉酶含量；Western印迹法检测肺组织中ROCK1、MYPT1、pMLC的相对表达量。结果：SAP模型组较对照组腹水量及腹水淀粉酶含量明显升高（t=-21.73、-40.72，均P<0.01）；SAP模型组、XBJ低剂量组、XBJ高剂量组腹水量及腹水淀粉酶含量逐渐降低（F=39.66、141.78，均P<0.01）。SAP模型组较对照组，胰腺干湿重比值、肺脏干湿重比值明显降低（t=19.83、15.47，均P<0.01），肺组织中Evans blue渗出量明显增高（t=-27.9，P<0.01）；SAP模型组、XBJ低剂量组、XBJ高剂量组胰腺干湿重比值、肺脏干湿重比值逐渐升高（F=75.19、15.47，均P<0.01），肺组织中Evans blue渗出量逐渐降低（F=99.52，P<0.01）。对照组大鼠胰腺组织结构完整，SAP模型组大鼠胰腺病理得分明显升高（t=-42.79，P<0.01）；XBJ低剂量组、XBJ高剂量组大鼠胰腺病理得分逐渐降低（F=175.43，P<0.01）。对照组大鼠肺组织结构完整，SAP模型组大鼠病理得分明显升高（t=-37.57，P<0.01）；XBJ低剂量组、XBJ高剂量组大鼠肺组织病变减轻，病理得分逐渐降低（F=126.00，P<0.01）。SAP模型组较对照组肺组织中ROCK1、pMLC蛋白表达量明显升高（t=-16.97、-13.53，均P<0.01），MYPT1表达量显著降低（t=23.30，P<0.01）；SAP模型组、XBJ低剂量组、XBJ高剂量组肺组织中ROCK1、pMLC蛋白表达量逐渐降低（F=84.89、50.84，均P<0.01），MYPT1表达量显著升高（F=48.68，P<0.01）。结论：XBJ通过降低多种细胞因子及DAMPs形成，抑制ROCK1-MYPT1-pMLC信号通路活化，对SAP肺损伤起到治疗的作用。

Abstract:

Objective：To investigate the therapeutic mechanism of Xuebijing（XBJ） injection on lung injury model of severe acute pancreatitis（SAP） in rats. Methods：A total of 80 male Spruge Dawley（SD） rats were randomly divided into 4 groups：control group，SAP model group，XBJ low-dose treatment group and XBJ high-dose treatment group，with 20 rats in each group. Except for the control group，the rats in all groups were injected with 4.5% sodium taurocholate solution（0.1 mL/100 g，0.05 mL/min） at uniform speed retrograde through the pancreatic bile duct to induce SAP. 30 min after successful molding，the rats in the low-dose and high-dose XBJ treatment groups were injected with XBJ injection through the tail vein（the dose was 5 and 20 mL/kg，respectively）. Control group and model group were given equal volume normal saline. After 24 h，10 rats were randomly selected from each group，and Evans blue was injected into the tail vein to detect the capillary permeability of lung tissue. The remaining rats were killed and their volume was measured with ascites. The dry-wet weight ratio was calculated by taking part of pancreas and lung tissues. Hematoxylin-eosin（HE） staining was used to observe the pathological changes of pancreas and lung，and pathological score was performed. The amylase content of ascites was detected by enzyme-linked immunosorbent assay（ELISA）. The relative expression levels of ROCK1，MYPT1 and pMLC in lung tissues were detected by Western blotting. Results：Compared with the control group，the water volume and amylase content of ascites in SAP model group were significantly increased（t=-21.73，-40.72，both P<0.01）. Compared with SAP model group，XBJ low-dose group and XBJ high-dose group，the water volume and amylase content of ascites gradually decreased（F=39.66，141.78，both P<0.01）. Compared with the control group，the dry-wet weight ratio of pancreas and dry-wet weight ratio of lung were significantly decreased in SAP model group（t=19.83，15.47，both P<0.01），and the Evans blue exudation amount in lung tissue was significantly increased（t=-27.9，P<0.01）. Compared with SAP model group，XBJ low-dose group and XBJ high-dose group，the dry-wet weight ratio of pancreas and dry-wet weight ratio of lung were gradually increased（F=75.19，15.47，both P<0.01），and the Evans blue exudation amount in lung tissue was gradually decreased（F=99.52，P<0.01）. The pancreatic tissue structure of the control group was intact，and the pancreatic pathological score of the SAP model group was significantly increased（t=-42.79，P<0.01）. The pathological scores of the pancreas in the low-dose group and the high-dose group of XBJ decreased gradually（F=175.43，P<0.01）. The lung structure of the control group was intact，and the pathological score of the SAP model group was significantly increased（t=-37.57，P<0.01）. Compared with the low-dose group of XBJ and the high-dose group of XBJ，the lung tissue lesions were reduced，and the pathological scores were decreased gradually（F=126.00，P<0.01）. Compared with the control group，the expression levels of ROCK1 and pMLC protein in SAP model group were significantly increased（t=-16.97，-13.53，both P<0.01），and the expression level of MYPT1 was significantly decreased（t=23.30，P<0.01）. Compared with SAP model group，XBJ low-dose group and XBJ high-dose group，the expression levels of ROCK1 and pMLC protein in lung tissue gradually decreased（F=84.89，50.84，both P<0.01），while the expression levels of MYPT1 were significantly increased（F=48.68，P<0.01）. Conclusion：XBJ plays a therapeutic role in SAP lung injury by reducing the formation of various cytokines and DAMPs，inhibiting the activation of ROCK1-MYPT1-pMLC signaling pathway.

参考文献/References:

[1] 中华医学会外科学分会胰腺外科学组.中国急性胰腺炎诊治指南（2021）[J].浙江实用医学，2021，26（6）：511-519，535.
[2] PEERY A F，CROCKETT S D，MURPHY C C，et al. Burden and cost of gastrointestinal，liver，and pancreatic diseases in the United States：update 2021[J]. Gastroenterology，2022，162（2）：621-644.
[3] WILEY M B，MEHROTRA K，BAUER J，et al. Acute pancreatitis：current clinical approaches，molecular pathophysiology，and potential therapeutics[J]. Pancreas，2023，52（6）：e335-e343.
[4] ZHANG Q，RAOOF M，CHEN Y，et al. Circulating mitochondrial DAMPs cause inflammatory responses to injury[J]. Nature，2010， 464（7285）：104-107.
[5] WANG Z，LIU J，WANG Y，et al. Identification of key biomarkers associated with immunogenic cell death and their regulatory mechanisms in severe acute pancreatitis based on WGCNA and machine learning[J]. Int J Mol Sci，2023，24（3）：3033.
[6] SHAH J，RANA S S. Acute respiratory distress syndrome in acute pancreatitis[J]. Indian J Gastroenterol，2020，39（2）：123-132.
[7] STERNBY H，HARTMAN H，THORLACIUS H，et al. The initial course of IL-1β，IL-6，IL-8，IL-10，IL-12，IFN-γ and TNF-α with regard to severity grade in acute pancreatitis[J]. Biomolecules，2021，11（4）：591.
[8] MEYER N J，GATTINONI L，CALFEE C S. Acute respiratory distress syndrome[J]. Lancet，2021，398（10300）：622-637.
[9] ZHOU J，ZHOU P，ZHANG Y，et al. Signal pathways and markers involved in acute lung injury induced by acute pancreatitis[J]. Dis Markers，2021，21（3）：9947.
[10] GOECKELER Z M，WYSOLMERSKI R B. Myosin light chain kinase-regulated endothelial cell contraction：the relationship between isometric tension，actin polymerization，and myosin phosphorylation[J]. J Cell Biol，1995，130（3）：613-627.
[11] ZHAO L，HU J，ZHENG P，et al. PAR1 regulates sepsis-induced vascular endothelial barrier dysfunction by mediating ERM phosphorylation via the RhoA/ROCK signaling pathway[J]. Int Immu-nopharmacol，2023，124：110992.
[12] WANG T，KANG W，DU L，et al. Rho-kinase inhibitor Y-27632 facilitates the proliferation，migration and pluripotency of human periodontal ligament stem cells[J]. J Cell Mol Med，2017，21：3100-3112.
[13] CHEN Y，TONG H，PAN Z，et al. Xuebijing injection attenuates pulmonary injury by reducing oxidative stress and proinflammatory damage in rats with heat stroke[J]. Exp Ther Med，2017，13（6）：3408-3416.
[14] LUO Z，CHEN W，XIANG M，et al. The preventive effect of Xuebijing injection against cytokine storm for severe patients with COVID-19：a prospective randomized controlled trial[J]. Eur J Integr Med，2021，42：101305.
[15] SONG Y，Yao C，YAO Y，et al. Xuebijing injection versus placebo for critically ill patients with severe community-acquired pneumonia：arandomized controlled trial[J]. Crit Care Med，2019，47（9）：e735-e743.
[16] 肖懿，冯志乔，张桂贤，等.血必净注射液调节线粒体N-甲酰肽/NLRP3炎症通路对重症急性胰腺炎大鼠模型的治疗机制[J].中国实验方剂学杂志，2022，28（7）：88-94.
[17] RADU M，CHERNOFF J. An in vivo assay to test blood vessel permeability[J]. J Vis Exp，2013，（73）：e50062.
[18] 胡炜. 清胰汤调控DAMPs对SAP-ALI大鼠的治疗机制及对肠道微生态调整的初步探讨[D].天津医科大学，2020.
[19] 肖懿，张桂贤，高瑞芳，等.重症急性胰腺炎大鼠血浆中6种线粒体N-甲酰肽及胰腺FPR1的表达研究[J].天津医药，2022，50（2）：150-154.
[20] WENCESLAU C F，MCCARTHY C G，WEBB RC. Formyl peptide receptor activation elicits endothelial cell contraction and vascular leakage[J]. Front Immunol，2016，7：297.
[21] YE L，LIAN Z Y，YANG H，et al. TNF-α activates RhoA/ROCK signaling pathway and increases permeability of endothelial cells infected with Listeria monocytogenes[J]. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi，2020，36（3）：193-197.
[22] YANG L，TANG L，DAI F，et al. Raf-1/CK2 and RhoA/ROCK signaling promote TNF-α-mediated endothelial apoptosis via regulating vimentin cytoskeleton[J]. Toxicology，2017，389：74-84.
[23] ZHAO H，KONG H，WANG W，et al. High glucose aggravates retinal endothelial cell dysfunction by activating the RhoA/ROCK1/pMLC/Connexin43 signaling pathway[J]. Invest Ophthalmol Vis Sci，2022，63（8）：22.
[24] MACDONALD J A，WALSH M P. Regulation of smooth muscle myosin light chain phosphatase by multisite phosphorylation of the myosin targeting subunit，MYPT1[J]. Cardiovasc Hematol Disord Drug Targets，2018，18（1）：4-13.
[25] FU W，LIU S，LUO L，et al. Anti-inflammatory mechanism of ulinastatin：inhibiting the hyperpermeability of vascular endothelial cells induced by TNF-α via the RhoA/ROCK signal pathway[J]. Int Immunopharmacol，2017，46：220-227.
[26] 刘昌孝，张铁军.基于“物质-药代-功效”关联的中药创新研发思路[J]. 中草药，2022，53（1）：1-7.
[27] 蔡楠，曹宁宁，赵利斌，等.基于“五原则”的血必净注射液质量标志物的预测分析[J]. 天津中医药，2021，38（8）：1062-1070.
[28] LIU L，CUI Q，SONG J，et al. Hydroxysafflower yellow A inhibits vascular adventitial fibroblast migration via NLRP3 inflammasome inhibition through autophagy activation[J]. Int J Mol Sci，2022，24（1）：172.
[29] YANG J，WANG R，CHENG X，et al. The vascular dilatation induced by Hydroxysafflower yellow A（HSYA） on rat mesenteric artery through TRPV4-dependent calcium influx in endothelial cells[J]. J Ethnopharmacol，2020，256：112790.
[30] ZHAO Y，SHAO C，ZHOU H，et al. Salvianolic acid B inhibits atherosclerosis and TNF-α-induced inflammation by regulating NF-κB/NLRP3 signaling pathway[J]. Phytomedicine，2023，119：155002.
[31] 庞彬彬，陈震，邢怡桥. 芍药苷调节RhoA/ROCK信号通路对实验性自身免疫性葡萄膜炎小鼠Th17/Treg免疫平衡的影响[J]. 中药新药与临床药理，2024，35（4）：506-512.
[32] 张育贵，张淑娟，边甜甜，等.芍药苷药理作用研究新进展[J]. 中草药，2019，50（15）：3735-3740.
[33] 李青泉，万建波，赵璐. 洋川芎内酯类化合物药理活性研究进展[J]. 针灸和草药（英文），2023，3（3）：180-188.

相似文献/References:

[1]刘 琪,王大为,陈 兴,等.氢气对吸入性损伤大鼠急性肺损伤的保护作用及量效研究[J].天津医科大学学报,2013,19(06):445.
　LIU Qi,WANG Da-wei,CHEN Xing,et al.Protective effects of hydrogen gas on acute lung injury induced by smoke inhalation in rats and its dose-effect relationship[J].Journal of Tianjin Medical University,2013,19(05):445.
[2]查玉杰,曹丽睿,何庆.基于网络药理学探讨三七抗急性肺损伤相关分子机制[J].天津医科大学学报,2021,27(05):446.
　ZHA Yu-jie,CAO Li-rui,HE Qing.Molecular mechanism of Panax notoginseng against acute lung injury based on network pharmacology[J].Journal of Tianjin Medical University,2021,27(05):446.
[3]齐颖,陈兵.盐酸右美托咪定对脓毒症急性肺损伤小鼠基质金属蛋白酶7抑制作用研究[J].天津医科大学学报,2022,28(01):65.
　QI Ying,CHEN Bing.Inhibitory effect of dexmedetomidine on serum matrix metalloproteinase 7 in mice with septic acute lung injury[J].Journal of Tianjin Medical University,2022,28(05):65.

备注/Memo

备注/Memo:

基金项目 天津市科技计划项目（21JCZDJC01220）；国家自然科学基金项目（82304797）；天津市科技计划项目（21JCYBJC01680）；天津市卫生健康科技项目（TJWJ2022MS049）
作者简介 王舜（1994-），男，主治医师，硕士在读，研究方向：中西医结合防治重症急性胰腺炎；通信作者：刘洪斌，E-mail：jtss@sina.com。

常用功能

更新日期/Last Update: 2024-09-20