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基于Wnt/β-catenin信号通路探讨清胰汤对重症急性胰腺炎肺泡上皮细胞损伤的修复机制

  • 王茵1,2

    机 构：

    1. 大连医科大学附属第一医院（大连 116000 ）

    2. 西安交通大学第一附属医院麻醉手术部（西安 710061 ）

    ×
  • ，白卜月1

    机 构：

    1. 大连医科大学附属第一医院（大连 116000 ）

    ×
  • ，陈海龙1,3

    机 构：

    1. 大连医科大学附属第一医院（大连 116000 ）

    3. 大连医科大学中西医结合研究院（大连 116000 ）

    ×
  • ，闻庆平1

    机 构：

    1. 大连医科大学附属第一医院（大连 116000 ）

    ×

1. 大连医科大学附属第一医院（大连 116000 ）；
2. 西安交通大学第一附属医院麻醉手术部（西安 710061 ）；
3. 大连医科大学中西医结合研究院（大连 116000 ）；

Based on wnt/β-catenin signaling pathway to explore mechanism of Qingyi decoction in promoting repair of alveolar epithelial cell damage in severe acute pancreatitis

  • Wang Yin1,2

    Affiliation：

    1. Department of Anesthesiology, The First Affiliated Hospital of Dalian Medical University, Dalain ( 116000 ), China

    ×
  • ，Bai Bu-yue1

    Affiliation：

    1. Department of Anesthesiology, The First Affiliated Hospital of Dalian Medical University, Dalain ( 116000 ), China

    ×
  • ，Chen Hai-long1,3

    Affiliation：

    1. Department of Anesthesiology, The First Affiliated Hospital of Dalian Medical University, Dalain ( 116000 ), China

    ×
  • ，Wen Qing-ping1

    Affiliation：

    1. Department of Anesthesiology, The First Affiliated Hospital of Dalian Medical University, Dalain ( 116000 ), China

    ×

Department of Anesthesiology, The First Affiliated Hospital of Dalian Medical University, Dalain ( 116000 ), China；

通讯作者:

闻庆平，E-mail: qingping_wen@yahoo.com

中图分类号:R657.5；R289

文献标识码:A

DOI:10.3969/j.issn.1007-6948.2024.04.010

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目录contents

    摘要

    目的：探究“Wnt/β-catenin”信号通路在重症急性胰腺炎（SAP）肺损伤发病机制中的作用及清胰汤（QYD）的干预机制。方法：将40只大鼠分为Sham组、假手术给药（Sham+QYD）组、SAP组和QYD治疗（SAP+QYD）组。采用经胆胰管逆行注射3.5%牛磺胆酸钠建立SAP大鼠模型，造模48 h后取胰腺和肺组织，苏木素-伊红（HE）染色检测组织病理损伤，真空干燥法检测肺湿/干重比值（W/D），ELISA法检测血清白细胞介素（IL）-6水平，TUNEL试剂盒和蛋白免疫印迹法（WB）检测肺组织凋亡情况。免疫荧光法检测CyclinD1、β-catenin与SFTPC双染情况。WB法检测肺组织中Wnt/β-catenin信号通路关键蛋白和下游细胞周期蛋白的表达。结果：与Sham组相比，SAP组大鼠HE切片显示胰腺和肺部病变严重，IL-6含量明显升高，SFTPC表达减少，提示肺泡上皮细胞严重损伤。与SAP大鼠相比，QYD治疗后大鼠胰腺和肺组织病理评分、W/D等各项指标均有不同程度改善，IL-6含量减少，凋亡蛋白Bcl-2/Bax及Wnt/β-catenin信号通路关键蛋白和下游细胞周期蛋白表达明显上调，免疫荧光双染结果显示QYD治疗组出现更多的双染细胞，差异均有统计学意义(P<0.05）。结论：QYD通过激活Wnt/β-catenin信号通路减少肺泡上皮细胞凋亡，进而在促进急性胰腺炎相关肺损伤修复中发挥关键作用。

    Abstract

    Objective To explore the role of "Wnt/β-catenin" signaling pathway in the pathogenesis of lung injury in severe acute pancreatitis (SAP) and the intervention effect of Qingyi decoction (QYD). Methods 40 rats were divided into Sham group, sham operation group (Sham+QYD), SAP group, and QYD treatment group (SAP+QYD). The SAP model was established by retrograde injection of 3.5% sodium taurocholate through the biliopancreatic duct. After 48 hours of modeling, pancreatic and lung tissues were taken, and tissue pathological damage was detected by hematoxylin-eosin (HE) staining, the lung wet/dry weight ratio (W/D) was measured using vacuum drying method, serum IL-6 levels were detected using enzyme-linked immunosorbent assay (ELISA), and lung tissue apoptosis was detected using TUNEL assay and Western blot (WB). Immunofluorescence assay was used to detect double staining of CyclinD1 and SFTPC, β-catenin and SFTPC. WB method was used to detect the expression of key proteins in the "Wnt/β-catenin" signaling pathway and downstream cyclins. Results Compared with the Sham group, HE slices of SAP group rats showed severe pancreatic and pulmonary lesions, significantly increased serum IL-6 levels, and decreased SFTPC expression, indicating severe damage to alveolar epithelial cells. Compared with SAP rats, QYD treatment showed varying degrees of improvement in various indicators such as pancreatic and lung tissue pathological scores, W/D, IL-6 content, apoptotic proteins Bcl-2/Bax, and the expression of key proteins in the Wnt/ β-catenin signaling pathway and downstream cyclins was significantly upregulated. The immunofluorescence double staining results showed that the QYD treatment group had more double stained cells, and the above differences were statistically significant (P<0.05). Conclusion QYD through activate Wnt/β-catenin signaling pathway plays a crucial role in promoting the repair of acute pancreatitis related lung injury by reducing apoptosis of alveolar epithelial cells.

  • 急性肺损伤（ALI）是SAP最常见且最严重的早期并发症[1]，SAP所致全身炎症反应过程中释放的炎性介质导致肺泡上皮细胞广泛损伤，富含蛋白的水肿液渗入肺间质和肺泡腔，进而引发急性肺水肿、肺不张及肺通气功能障碍[2]。肺泡上皮作为人体重要的屏障之一，主要由肺泡I型上皮细胞（AEC I）和含有表面活性物质的肺泡Ⅱ型上皮细胞（AEC Ⅱ）组成。作为“肺泡上皮祖细胞”的AEC Ⅱ[3]，不仅能够通过有丝分裂在基底膜上增殖进行自我更新，而且能够分化为AEC I，恢复肺泡正常的屏障结构和气体交换等功能[4-5]。因此，肺损伤后加快促进AEC II增殖和分化为AEC I以修复损伤肺泡上皮结构功能，已成为治疗ALI的有效策略。

  • 清胰汤是经多年临床实践和动物实验证实治疗SAP的有效方剂[6-7]，由大柴胡汤和大承气汤加减化裁而成，有疏肝理气、清热利湿、通腑化滞等功效[8]。研究表明，QYD在治疗SAP及相关胰腺、肠黏膜、肺组织等脏器损伤中发挥重要作用[9-10]。然而，QYD能否促进损伤后AEC Ⅱ的修复尚不明确。经典的Wnt/β-catenin信号通路激活会诱导β-catenin的积累和向细胞核的转移，从而促进参与细胞增殖、凋亡、分化和迁移的基因的表达[11]，参与组织稳态、迁移、侵袭、凋亡和增殖过程[12-13]，还参与损伤恢复和分化等生理活动[14-16]。最近在多种模型的实验中证实，Wnt/β-catenin信号通路在肺再生过程中能够促进AEC Ⅱ增殖并抑制分化[17]，但其调控AEC Ⅱ促进肺损伤修复的确切机制尚不清楚。本研究构建SAP大鼠模型，基于Wnt/β-catenin信号通路探究清胰汤促进重症急性胰腺炎肺泡上皮细胞损伤修复的作用机制，旨在为临床上应用清胰汤防治急性胰腺炎肺损伤（APALI）提供理论依据和新的治疗策略。

  • 1 材料与方法

  • 1.1 实验动物

  • 雄性SD大鼠50只，6~8周龄，体质量180~220g，由中国大连人类疾病基因组工程动物模型研究所提供[动物实验伦理审查表编号（AEE19003），实验动物质量合格证编号（110322221100410668），动物实验设施合格证编号（00123317）]。

  • 1.2 实验药物以及试剂

  • 清胰汤由同仁堂药房提供，方剂组成：柴胡15 g，黄芩12 g，白芍 15 g，木香15 g，元胡15 g，栀子15 g，大黄20 g，芒硝10 g。除大黄、芒硝外，其他生药饮片加入10倍量的水，浸泡过夜后，沸水煎煮1 h后收集药液。再加8倍水煮沸0.5 h后加入大黄继续煮0.5 h，收集药液。将2次收集的药液混在一起，冷却后加入芒硝，旋蒸并收集药液。浓缩至1 g/mL，4℃保存。Wnt3a、SFTPC（Proteintech公司）；β-Catenin、Cyclin D1、Bcl-2、Bax、β-actin（HUABIO公司）；TUNEL试剂盒（Roche公司）；大鼠淀粉酶及白细胞介素（IL）-6 ELISA试剂盒（上海西唐生物技术公司）；HE染液套装（北京索莱宝公司）。

  • 1.3 主要仪器设备

  • 组织研磨机（上海净信实业发展有限公司）；动物实验手术器械（上海医疗器械集团有限公司）；台式低温高速离心机（美国Thermo公司）；倒置荧光相差显微镜（日本OLYMPUS公司）；病理切片机（上海徕卡仪器有限公司）；凝胶成像仪（美国Thermo公司）。

  • 1.4 动物实验

  • 选择40只雄性SD大鼠，随机分成4组：Sham组、Sham+QYD组、SAP组、SAP+QYD组，每组10只。选取SAP组和SAP+QYD组大鼠，给予腹腔注射1%戊巴比妥钠溶液麻醉后备皮，仰卧固定消毒后，于腹壁行1~2 cm正中切口，找出十二指肠和胆胰管后夹闭胆管，采用30 G注射器针头由十二指肠乳头端逆行穿入胆胰管，30 s内均匀缓慢注射3.5%牛磺胆酸钠溶液（1 mL/kg），见到胰腺组织充血水肿后拔除针头，复位十二指肠并缝合关腹。对于Sham组和Sham+QYD组大鼠，只需将3.5%牛磺胆酸钠溶液替换成无菌生理盐水，其他步骤完全一致。Sham+QYD组和SAP+QYD于造模后4 h给予清胰汤（浓度5.27 mL/kg），之后每12 h给予1次，Sham组和SAP组给予等量生理盐水。

  • 1.5 大鼠胰腺和肺HE染色及病理评分

  • 将固定好的胰腺和肺组织于48 h内乙醇梯度脱水，石蜡包埋，切片机将其切成4 μm厚的薄片，切片经HE染色后用显微镜观察，进行胰腺、肺组织病理学评分[18]。

  • 1.6 大鼠肺肺湿干比和炎性因子IL-6检测

  • 大鼠放血处死后，取右肺下叶组织标本，记录电子秤所称湿质量，随后置于80℃烤箱中24 h并记录干质量；计算肺湿干比（W/D）=（湿质量－干质量）/湿质量×100%。取大鼠腹主动脉血，静置1 h后，4℃离心10 min（3000 r/min），回收血清并放入-80℃冰箱备用。采用ELISA试剂盒，于450 nm处测光密度（OD）值，采用标准曲线求IL-6浓度。

  • 1.7 TUNEL分析肺组织凋亡情况

  • TUNEL检测采用Roche试剂盒进行。石蜡组织切片步骤同前1.5大鼠胰腺和肺HE染色及病理评分部分。随后将组织切片置于修复盒，微波炉进行抗原修复。再经血清封闭后将试剂盒内的试剂1（TdT）和试剂2（dUTP）按特定比例1∶10混合后滴加在切片上，并将切片平放于加少量水的湿盒内，4℃孵育过夜。DAPI复染细胞核后封片，于显微镜下观察并采集图像。

  • 1.8 免疫荧光法检测肺组织SFTPC的表达及CyclinD1、SFTPC、β-catenin和SFTPC双染情况

  • 将右肺中叶用4%多聚甲醛保存，石蜡浸泡，切成4 μm厚。而后进行脱蜡、抗原修复和血清封闭，给予SFTPC、偶联CyclinD1的SFTPC和偶联β-catenin的SFTPC一抗并置于湿箱中4℃孵育过夜。PBS冲洗3次后，室温下二抗孵育切片1 h，应用DAPI孵育10 min染细胞核，使用尼康倒置荧光显微镜观察荧光表达和共定位。

  • 1.9 蛋白免疫印迹法（WB）检测肺组织中Wnt3a、β-catenin、CyclinD1、SFTPC、Bax和Bcl-2的表达

  • 利用RIPA缓冲液从肺组织中提取总蛋白。变性后，所有样品中等量的蛋白质在SDS-PAGE凝胶上电泳并传输到NC膜上。用5%脱脂牛奶在室温下封闭2 h，4℃一抗孵育过夜，室温二抗孵育1 h。ECL程序检测肺组织中相关蛋白条带的表达，使用Image J软件对各组蛋白进行灰度分析。

  • 1.10 统计学分析

  • 使用GraphPad Prism 8分析结果。正态分布结果以均数±标准差表示。采用单因素方差分析（ANOVA）和t检验进行数据分析，P<0.05为差异具有统计学意义。

  • 2 结果

  • 2.1 各组大鼠胰腺和肺病理改变对比

  • HE染色显示，与SAP组比较，使用QYD治疗后，胰腺腺泡的明显坏死、小叶结构的严重破坏、广泛的出血和水肿以及炎性细胞的大量浸润均有所缓解；广泛的肺泡壁增厚、肺泡间隔增宽、肺间质水肿和炎细胞浸润等肺病理改变也有明显改善。两组胰腺组织和肺组织镜下病理评分比较，差异存在统计学意义（P<0.001，见图1）。

  • 2.2 各组大鼠肺W/D和炎性因子IL-6含量比较

  • 与Sham组相比，SAP组大鼠肺W/D显著升高，经QYD治疗后明显回落。SAP组血清IL-6水平明显高于Sham组，经治疗后IL-6水平明显降低（见图2）。

  • 图1 各组大鼠胰腺和肺病理改变比较

  • 图2 各组大鼠肺W/D和血清IL-6含量比较

  • 2.3 清胰汤抑制SAP所致的肺泡上皮细胞凋亡

  • 与Sham组相比，SAP组凋亡小体明显增加，而QYD能够减少凋亡的发生。WB结果表明，SAP组抗凋亡相关蛋白Bcl-2表达下调，促凋亡相关蛋白Bax表达上调，而QYD治疗能够部分逆转上述变化，差异有统计学意义（图3）。

  • 图3 清胰汤抑制SAP情况下肺泡上皮细胞的凋亡

  • 2.4 清胰汤促进SAP肺泡上皮细胞的修复

  • 免疫荧光显示，SAP组SFTPC阳性细胞数量显著减少，QYD治疗后明显反弹。WB检测结果显示显示，SAP组SFTPC以及CyclinD1蛋白表达明显降低，QYD上调了SFTPC及CyclinD1的表达，SFTPC与CyclinD1免疫荧光双染检测也有类似结果（图4）。

  • 2.5 清胰汤通过激活Wnt/β-catenin信号通路来促进肺泡上皮细胞的修复

  • 与Sham组相比，SAP组Wnt3a和β-catenin水平明显降低，而QYD能够上调Wnt3a和β-catenin的表达。免疫荧光双染色法鉴定SFTPC和β-catenin双阳性细胞，显示SAP组SFTPC和β-catenin双阳性细胞数量显著降低，经QYD治疗后双阳性细胞数量显著升高（图5）。

  • 图4 清胰汤促进SAP情况下肺泡上皮细胞的修复

  • 图5 清胰汤通过激活Wnt/β-catenin信号通路来促进肺泡上皮细胞的修复

  • 3 讨论

  • 肺损伤是SAP严重且普遍的胰外并发症，可进一步发展为急性呼吸窘迫综合征，是SAP患者主要死亡原因之一[19-20]。肺泡上皮细胞是APALI发生过程中炎症损伤的主要部位[21]。其中，AEC Ⅱ在ALI发展中起到重要作用[22]，能够参与肺泡上皮的修复过程[23-24]。中医理论认为“肺与大肠相表里”，QYD具有疏肝理气、清热利湿、通腑化滞的功效[25]，可有效治疗SAP及相关性肺损伤，但其具体机制尚不清楚。因此，本研究我们构建了SAP大鼠模型，试图探讨QYD在SAP中对 AEC Ⅱ的调节作用。结果表明，SAP肺气血屏障损伤严重，AEC Ⅱ标记蛋白SFTPC显著下调，而QYD能够显著减弱炎症反应，改善肺呼吸功能，抑制AEC Ⅱ细胞凋亡，促进AEC Ⅱ细胞增殖，进而修复受损的肺泡上皮。

  • 研究发现，中药和中药单体可通过促进AEC Ⅱ细胞增殖和抑制细胞凋亡来减轻损伤。另有研究表明，白藜芦醇可以通过激活SIRT1/PGC-1a信号通路减少肺泡上皮细胞凋亡，缓解高氧诱导的肺损伤[26]。另一项研究报道，黄芩苷可能通过增加miR-200b-3p通路的表达来抑制ERK/JNK炎症，进而抑制炎症相关蛋白的表达，促进AEC Ⅱ增殖[27]。清胰汤是经多年临床实践和动物实验证实能，治疗急性胰腺炎的有效方剂。有研究发现，QYD通过抑制炎症反应和肺组织的凋亡来缓解肺损伤[28]。，Sun等人[29]报道QYD可抑制AEC Ⅱ细胞凋亡。本课题组前期研究证实，大黄素通过降低CIRP表达，抑制大鼠肺部炎症反应，减弱中性粒细胞浸润，减轻肺组织损伤[30]。推测QYD可能抑制胰腺源性损伤或氧化应激以及炎症因子，如CIRP，抑制AEC Ⅱ细胞凋亡并促进其增殖[30-31]。为了进一步证实这一假设，本研究我们检测了肺气血屏障的完整性和AEC Ⅱ标记蛋白SFTPC，结果表明，SAP的肺泡上皮细胞完整性受损，SFTPC表达降低。同时，QYD可显著修复肺气血屏障的完整性，特别是促进AEC Ⅱ的修复，有效改善肺呼吸功能。

  • 既往研究显示，Wnt/β-catenin信号通路激活可促进AEC Ⅱ存活、迁移以及向AEC I分化，进而缓解肺损伤[32]。此外，在小鼠模型中敲低RAB6基因可激活Wnt/β-catenin信号通路，在减轻肺损伤和延缓肺纤维化中起关键作用[33]。在AP所致ALI中，Wnt/β-catenin通路激动剂Wnt3a一方面减少中性粒细胞进入肺泡上皮细胞，减轻肺部炎症[34]；另一方面，它减少了P2X7R或LPS介导的肺泡上皮细胞的死亡，进而促进了肺气血屏障结构和功能的恢复[35]。然而，Wnt/β-catenin信号通路在急性胰腺炎肺损伤发病机制中的确切途径尚不清楚。本研究旨在探索QYD在APALI期间促进AEC Ⅱ恢复的作用是否与Wnt/β-catenin信号通路相关，结果表明，Wnt信号通路的关键蛋白Wnt3a和β-catenin在APALI大鼠肺组织中表达降低，而QYD治疗可以显著提升Wnt3a和β-catenin的表达，并均与SFTPC共定位。这些都说明了QYD减轻APALI的严重程度是通过促进Wnt3a的表达，激活Wnt/β-catenin信号通路，实现抗AEC Ⅱ的凋亡和促进肺组织修复而起作用的。

  • 综上所述，SAP过程中AEC Ⅱ细胞凋亡增加，肺气血屏障被破坏，QYD通过促进Wnt/β-catenin信号通路抑制AEC Ⅱ的凋亡并且促进其修复，从而修复受损的肺气血屏障，缓解APALI。本研究也有一定局限，QYD作为中药是多角度、多靶点发挥作用的，其抑制AEC Ⅱ凋亡的具体机制还有待于进一步深入地研究。

  • 参考文献

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    • [2] Akbarshahi H,Rosendahl AH,Westergren-Thorsson G,et al.Acute lung injury in acute pancreatitis:awaiting the big leap[J].Respir Med,2012,106(9):1199-1210.

    • [3] Jansing NL,McClendon J,Henson PM,et al.Unbiased quantitation of alveolar type II to alveolar type I cell transdifferentiation during repair after lung injury in mice[J].Am J Respir Cell Mol Biol,2017,57(5):519-526.

    • [4] Mutze K,Vierkotten S,Milosevic J,et al.Enolase 1(ENO1)and protein disulfide-isomerase associated 3(PDIA3)regulate Wnt/β-catenin-driven trans-differentiation of murine alveolar epithelial cells[J].Dis Model Mech,2015,8(8):877-890.

    • [5] Shen MX,Luo ZQ,Zhou Y.Regeneration-associated transitional state cells in pulmonary fibrosis[J].Int J Mol Sci,2022,23(12):6757.

    • [6] Zhang GX,Zhan C,Wang K,et al.Qingyi Decoction amerliorates acute biliary pancreatitis by targeting Gpbar1/NF-kb pathway[J].Front Biosci,2019,24(5):833-848.Wang GY,Shang D,Zhang GX,et al.Qingyi decoction attenuates intestinal epithelial cell injury via the calcineurin/nuclear factor of activated T-cells pathway[J].World J Gastroenterol,2022,28(29):3825-3837.

    • [7] Su S,Liang TC,Zhou X,et al.Qingyi decoction attenuates severe acute pancreatitis in rats via inhibition of inflammation and protection of the intestinal barrier[J].J Int Med Res,2019,47(5):2215-2227.

    • [8] Wei TF,Zhao L,Huang P,et al.Qing-yi decoction in the treatment of acute pancreatitis:an integrated approach based on chemical profile,network pharmacology,molecular docking and experimental evaluation[J].Front Pharmacol,2021,12:590994.

    • [9] Wang GY,Shang D,Zhang GX,et al.Effects of QingYi Decoction on inflammatory markers in patients with acute pancreatitis:a meta-analysis[J].Phytomedicine,2022,95:153738.

    • [10] Colozza G,Koo BK.Wnt/β-catenin signaling:structure,assembly and endocytosis of the signalosome[J].Dev Growth Differ,2021,63(3):199-218.

    • [11] Nusse R,Clevers H.Wnt/β-catenin signaling,disease,and emerging therapeutic modalities[J].Cell,2017,169(6):985-999.

    • [12] Aros CJ,Pantoja CJ,Gomperts BN.Wnt signaling in lung development,regeneration,and disease progression[J].Commun Biol,2021,4(1):601.

    • [13] Raslan AA,Yoon JK.WNT signaling in lung repair and regeneration[J].Mol Cells,2020,43(9):774-783.

    • [14] Steinhart Z,Angers S.Wnt signaling in development and tissue homeostasis[J].Development,2018,145(11):dev146589.

    • [15] Mohapatra P,Chandrasekaran N.Wnt/β-catenin targeting in liver carcinoma through nanotechnology-based drug repurposing:a review[J].Biomed Pharmacother,2022,155:113713.

    • [16] Nabhan AN,Brownfield DG,Harbury PB,et al.Single-cell Wnt signaling niches maintain stemness of alveolar type 2 cells[J].Science,2018,359(6380):1118-1123.

    • [17] Osman MO,Kristensen JU,Jacobsen NO,et al.A monoclonal anti-interleukin 8 antibody(WS-4)inhibits cytokine response and acute lung injury in experimental severe acute necrotising pancreatitis in rabbits[J].Gut,1998,43(2):232-239.

    • [18] Elder AS,Saccone GT,Dixon DL.Lung injury in acute pancreatitis:mechanisms underlying augmented secondary injury[J].Pancreatology,2012,12(1):49-56.

    • [19] Liang XY,Jia TX,Zhang M.Intestinal bacterial overgrowth in the early stage of severe acute pancreatitis is associated with acute respiratory distress syndrome[J].World J Gastroenterol,2021,27(15):1643-1654.

    • [20] Sun TH,Huang ZY,Zhang H,et al.TAZ is required for lung alveolar epithelial cell differentiation after injury[J].JCI Insight,2019,5(14):e128674.

    • [21] Hu C,Sun JH,Du J,et al.The Hippo-YAP pathway regulates the proliferation of alveolar epithelial progenitors after acute lung injury[J].Cell Biol Int,2019,43(10):1174-1183.

    • [22] Zhang HL,Cui Y,Zhou ZY,et al.Alveolar type 2 epithelial cells as potential therapeutics for acute lung injury/acute respiratory distress syndrome[J].Curr Pharm Des,2019,25(46):4877-4882.

    • [23] Dargaville PA,Kamlin COF,Orsini F,et al.Effect of minimally invasive surfactant therapy vs sham treatment on death or bronchopulmonary dysplasia in preterm infants with respiratory distress syndrome:the OPTIMIST-a randomized clinical trial[J].JAMA,2021,326(24):2478-2487.

    • [24] 郭颢雅,许才明,张赟,等.基于“和下清消”法运用清胰汤治疗急性胰腺炎的源流与临床应用[J].世界科学技术-中医药现代化,2022,24(11):4269-4278.

    • [25] Zhu XD,Wang F,Lei XP,et al.Resveratrol alleviates alveolar epithelial cell injury induced by hyperoxia by reducing apoptosis and mitochondrial dysfunction[J].Exp Biol Med,2021,246(5):596-606.

    • [26] Duan XY,Sun Y,Zhao ZF,et al.Baicalin attenuates LPS-induced alveolar type II epithelial cell A549 injury by attenuation of the FSTL1 signaling pathway via increasing miR-200b-3p expression[J].Innate Immun,2021,27(4):294-312.

    • [27] Liu GL,Zhang JW,Chen HL,et al.Effects and mechanisms of alveolar type II epithelial cell apoptosis in severe pancreatitis-induced acute lung injury[J].Exp Ther Med,2014,7(3):565-572.

    • [28] Sun ZW,Li L,Qu JL,et al.Proteomic analysis of therapeutic effects of Qingyi pellet on rodent severe acute pancreatitis-associated lung injury[J].Biomed Pharmacother,2019,118:109300.

    • [29] Xu QS,Wang MF,Guo HY,et al.Emodin alleviates severe acute pancreatitis-associated acute lung injury by inhibiting the cold-inducible RNA-binding protein(CIRP)-mediated activation of the NLRP3/IL-1β/CXCL1 signaling[J].Front Pharmacol,2021,12:655372.

    • [30] Xu CM,Zhang JY,Liu J,et al.Proteomic analysis reveals the protective effects of emodin on severe acute pancreatitis induced lung injury by inhibiting neutrophil proteases activity[J].J Proteomics,2020,220:103760.

    • [31] Flozak AS,Lam AP,Russell S,et al.Beta-catenin/T-cell factor signaling is activated during lung injury and promotes the survival and migration of alveolar epithelial cells[J].J Biol Chem,2010,285(5):3157-3167.

    • [32] Yang LW,Liu G,Li XM,et al.Small GTPase RAB6 deficiency promotes alveolar progenitor cell renewal and attenuates PM2.5-induced lung injury and fibrosis[J].Cell Death Dis,2020,11(10):827.

    • [33] Guo YJ,Mishra A,Howland E,et al.Platelet-derived Wnt antagonist Dickkopf-1 is implicated in ICAM-1/VCAM-1-mediated neutrophilic acute lung inflammation[J].Blood,2015,126(19):2220-2229.

    • [34] Guo Y,Mishra A,Weng T,et al.Wnt3a mitigates acute lung injury by reducing P2X7 receptor-mediated alveolar epithelial type I cell death[J].Cell Death Dis,2014,5(6):e1286.

图1 各组大鼠胰腺和肺病理改变比较

图2 各组大鼠肺W/D和血清IL-6含量比较

图3 清胰汤抑制SAP情况下肺泡上皮细胞的凋亡

图4 清胰汤促进SAP情况下肺泡上皮细胞的修复

图5 清胰汤通过激活Wnt/β-catenin信号通路来促进肺泡上皮细胞的修复

图表 1/1

  • 基本信息

    中图分类号: R657.5；R289
    文献标识码: A
    DOI: 10.3969/j.issn.1007-6948.2024.04.010

    基金信息

    辽宁省重点实验室项目（LJKZ0851）；

    稿件历史

    收稿日期: 2023-12-10

  • 参考文献

    • [1] Zhou JL,Zhou PC,Zhang YY,et al.Signal pathways and markers involved in acute lung injury induced by acute pancreatitis[J].Dis Markers,2021,2021:9947047.

    • [2] Akbarshahi H,Rosendahl AH,Westergren-Thorsson G,et al.Acute lung injury in acute pancreatitis:awaiting the big leap[J].Respir Med,2012,106(9):1199-1210.

    • [3] Jansing NL,McClendon J,Henson PM,et al.Unbiased quantitation of alveolar type II to alveolar type I cell transdifferentiation during repair after lung injury in mice[J].Am J Respir Cell Mol Biol,2017,57(5):519-526.

    • [4] Mutze K,Vierkotten S,Milosevic J,et al.Enolase 1(ENO1)and protein disulfide-isomerase associated 3(PDIA3)regulate Wnt/β-catenin-driven trans-differentiation of murine alveolar epithelial cells[J].Dis Model Mech,2015,8(8):877-890.

    • [5] Shen MX,Luo ZQ,Zhou Y.Regeneration-associated transitional state cells in pulmonary fibrosis[J].Int J Mol Sci,2022,23(12):6757.

    • [6] Zhang GX,Zhan C,Wang K,et al.Qingyi Decoction amerliorates acute biliary pancreatitis by targeting Gpbar1/NF-kb pathway[J].Front Biosci,2019,24(5):833-848.Wang GY,Shang D,Zhang GX,et al.Qingyi decoction attenuates intestinal epithelial cell injury via the calcineurin/nuclear factor of activated T-cells pathway[J].World J Gastroenterol,2022,28(29):3825-3837.

    • [7] Su S,Liang TC,Zhou X,et al.Qingyi decoction attenuates severe acute pancreatitis in rats via inhibition of inflammation and protection of the intestinal barrier[J].J Int Med Res,2019,47(5):2215-2227.

    • [8] Wei TF,Zhao L,Huang P,et al.Qing-yi decoction in the treatment of acute pancreatitis:an integrated approach based on chemical profile,network pharmacology,molecular docking and experimental evaluation[J].Front Pharmacol,2021,12:590994.

    • [9] Wang GY,Shang D,Zhang GX,et al.Effects of QingYi Decoction on inflammatory markers in patients with acute pancreatitis:a meta-analysis[J].Phytomedicine,2022,95:153738.

    • [10] Colozza G,Koo BK.Wnt/β-catenin signaling:structure,assembly and endocytosis of the signalosome[J].Dev Growth Differ,2021,63(3):199-218.

    • [11] Nusse R,Clevers H.Wnt/β-catenin signaling,disease,and emerging therapeutic modalities[J].Cell,2017,169(6):985-999.

    • [12] Aros CJ,Pantoja CJ,Gomperts BN.Wnt signaling in lung development,regeneration,and disease progression[J].Commun Biol,2021,4(1):601.

    • [13] Raslan AA,Yoon JK.WNT signaling in lung repair and regeneration[J].Mol Cells,2020,43(9):774-783.

    • [14] Steinhart Z,Angers S.Wnt signaling in development and tissue homeostasis[J].Development,2018,145(11):dev146589.

    • [15] Mohapatra P,Chandrasekaran N.Wnt/β-catenin targeting in liver carcinoma through nanotechnology-based drug repurposing:a review[J].Biomed Pharmacother,2022,155:113713.

    • [16] Nabhan AN,Brownfield DG,Harbury PB,et al.Single-cell Wnt signaling niches maintain stemness of alveolar type 2 cells[J].Science,2018,359(6380):1118-1123.

    • [17] Osman MO,Kristensen JU,Jacobsen NO,et al.A monoclonal anti-interleukin 8 antibody(WS-4)inhibits cytokine response and acute lung injury in experimental severe acute necrotising pancreatitis in rabbits[J].Gut,1998,43(2):232-239.

    • [18] Elder AS,Saccone GT,Dixon DL.Lung injury in acute pancreatitis:mechanisms underlying augmented secondary injury[J].Pancreatology,2012,12(1):49-56.

    • [19] Liang XY,Jia TX,Zhang M.Intestinal bacterial overgrowth in the early stage of severe acute pancreatitis is associated with acute respiratory distress syndrome[J].World J Gastroenterol,2021,27(15):1643-1654.

    • [20] Sun TH,Huang ZY,Zhang H,et al.TAZ is required for lung alveolar epithelial cell differentiation after injury[J].JCI Insight,2019,5(14):e128674.

    • [21] Hu C,Sun JH,Du J,et al.The Hippo-YAP pathway regulates the proliferation of alveolar epithelial progenitors after acute lung injury[J].Cell Biol Int,2019,43(10):1174-1183.

    • [22] Zhang HL,Cui Y,Zhou ZY,et al.Alveolar type 2 epithelial cells as potential therapeutics for acute lung injury/acute respiratory distress syndrome[J].Curr Pharm Des,2019,25(46):4877-4882.

    • [23] Dargaville PA,Kamlin COF,Orsini F,et al.Effect of minimally invasive surfactant therapy vs sham treatment on death or bronchopulmonary dysplasia in preterm infants with respiratory distress syndrome:the OPTIMIST-a randomized clinical trial[J].JAMA,2021,326(24):2478-2487.

    • [24] 郭颢雅,许才明,张赟,等.基于“和下清消”法运用清胰汤治疗急性胰腺炎的源流与临床应用[J].世界科学技术-中医药现代化,2022,24(11):4269-4278.

    • [25] Zhu XD,Wang F,Lei XP,et al.Resveratrol alleviates alveolar epithelial cell injury induced by hyperoxia by reducing apoptosis and mitochondrial dysfunction[J].Exp Biol Med,2021,246(5):596-606.

    • [26] Duan XY,Sun Y,Zhao ZF,et al.Baicalin attenuates LPS-induced alveolar type II epithelial cell A549 injury by attenuation of the FSTL1 signaling pathway via increasing miR-200b-3p expression[J].Innate Immun,2021,27(4):294-312.

    • [27] Liu GL,Zhang JW,Chen HL,et al.Effects and mechanisms of alveolar type II epithelial cell apoptosis in severe pancreatitis-induced acute lung injury[J].Exp Ther Med,2014,7(3):565-572.

    • [28] Sun ZW,Li L,Qu JL,et al.Proteomic analysis of therapeutic effects of Qingyi pellet on rodent severe acute pancreatitis-associated lung injury[J].Biomed Pharmacother,2019,118:109300.

    • [29] Xu QS,Wang MF,Guo HY,et al.Emodin alleviates severe acute pancreatitis-associated acute lung injury by inhibiting the cold-inducible RNA-binding protein(CIRP)-mediated activation of the NLRP3/IL-1β/CXCL1 signaling[J].Front Pharmacol,2021,12:655372.

    • [30] Xu CM,Zhang JY,Liu J,et al.Proteomic analysis reveals the protective effects of emodin on severe acute pancreatitis induced lung injury by inhibiting neutrophil proteases activity[J].J Proteomics,2020,220:103760.

    • [31] Flozak AS,Lam AP,Russell S,et al.Beta-catenin/T-cell factor signaling is activated during lung injury and promotes the survival and migration of alveolar epithelial cells[J].J Biol Chem,2010,285(5):3157-3167.

    • [32] Yang LW,Liu G,Li XM,et al.Small GTPase RAB6 deficiency promotes alveolar progenitor cell renewal and attenuates PM2.5-induced lung injury and fibrosis[J].Cell Death Dis,2020,11(10):827.

    • [33] Guo YJ,Mishra A,Howland E,et al.Platelet-derived Wnt antagonist Dickkopf-1 is implicated in ICAM-1/VCAM-1-mediated neutrophilic acute lung inflammation[J].Blood,2015,126(19):2220-2229.

    • [34] Guo Y,Mishra A,Weng T,et al.Wnt3a mitigates acute lung injury by reducing P2X7 receptor-mediated alveolar epithelial type I cell death[J].Cell Death Dis,2014,5(6):e1286.


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