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

《中华肺部疾病杂志》[ISSN:1006-6977/CN:61-1281/TN]

卷:

期数:

2023年02期

页码:

156-163

栏目:

论著

出版日期:

2023-04-20

文章信息/Info

Title:

Effect of human amniotic mesenchymal stem cell regulating MAPK signaling pathway of aquaporin 1 in acute lung injury

作者:

李埝 赵建军 张建勇 赵睿桢

563000 遵义,遵义医科大学附属医院呼吸与危重症医学科呼吸二病区

Author(s):

Li nian;  Zhao Jianjun;  Zhang Jianyong;  Zhao Ruizhen.

he Second Ward, Department of Respiratory and Critical Care Medicine, Affiliated Hospital of Zunyi Medical University, Zunyi 563000, China

关键词:

人羊膜间充质干细胞;  脂多糖;  急性肺损伤;  水通道蛋白 1

Keywords:

Human amniotic mesenchymal stem cells;  Lipopolysaccharide;  Acute lung injury;  Aquaporin 1

分类号:

R563

DOI:

10.3877/cma.j.issn.1674-6902.2023.02.002

摘要:

目的 分析人羊膜间充质干细胞(human amniotic mesenchymal stromal cells, hAMSCs)移植后对急性肺损伤(acute lung injury, ALI)大鼠肺组织水通道蛋白 1(AQP1)和磷酸化p38丝裂原活化蛋白激酶(p-p38MAPK)表达的影响及对肺损伤的作用机制。方法 随机将84只SD雄性大鼠分为4组(n=21):生理盐水对照组(NS组)、LPS诱导ALI模型组(LPS 组)、hAMSCs对照组(NH组)及hAMSCs干预组(LH组)。LPS组和LH组大鼠通过舌下静脉注射LPS(4 mg/kg)造模; NS组和NH组大鼠通过舌下静脉注射250 μl生理盐水,2 h后NH组和LH组大鼠通过尾静脉注射250 μl经 DAPI 标记的 hAMSCs(2.5×10⁶个); NS组和LPS组大鼠通过尾静脉注射250 μl生理盐水。每组分别于6 h、12 h、72 h随机处死7只大鼠观察hAMSCs在每组中的定植情况、肺组织病理形态,湿干重比值(W/D),Elisa法检测BALF中TNF-α、IL-1β水平,IHC及WB法检测各组大鼠肺组织 AQP1、p38MAPK和 p-p38 MAPK 的表达。结果 ①NH和LH组大鼠肺泡周围有蓝色荧光标记的hAMSCs, LH组分布较多,NH组分布较少; ②NS组、NH 组对应时相点的大鼠肺组织病理评分、W/D比值、BALF中TNF-α和IL-1β水平、AQP1和p-p38 MAPK蛋白表达量相比较无显著性差异(P>0.05); ③与 NS 组、NH 组时相点相比较,LPS组、LH组对应时相点大鼠肺组织病理评分、W/D比值、BALF中TNF-α、IL-1β水平和p-p38 MAPK 蛋白表达量升高,AQP1表达量降低(P<0.05); ④LPS 6 h组与12 h组大鼠肺组织病理评分、W/D 比值、BALF中TNF-α 和IL-1β水平、AQP1和p-p38 MAPK蛋白表达量无明显差异(P>0.05); 与LPS 6 h组、12 h组相比较,LPS 72 h组大鼠肺组织病理评分、W/D比值、BALF中IL-1β和TNF-α水平、p-p38 MAPK蛋白表达量降低,AQP1表达量升高(P<0.05); LH 6 h组与12 h组大鼠肺组织病理评分、W/D比值、BALF中IL-1β和TNF-α水平、AQP1和p-p38 MAPK蛋白表达量无明显差异(P>0.05); 与LH 6 h组、12 h组相比较,LH 72 h组大鼠肺组织病理评分、W/D比值、BALF中IL-1β和TNF-α水平、p-p38 MAPK蛋白表达量降低,AQP1表达量升高(P<0.05); LH组与LPS组时相点相比,大鼠肺组织病理评分、W/D比值、BALF中TNF-α和IL-1β水平、p-p38 MAPK蛋白表达量降低,AQP1表达量升高(P<0.05); ⑤每组时相点大鼠肺组织p38 MAPK表达量相比较无明显差异(P>0.05)。结论 ①外源性hAMSCs能归巢并定植于受损伤肺组织; ②hAMSCs可抑制p38 MAPK激活,减少炎症因子TNF-α、IL-1β释放,减轻肺部炎症反应; ③hAMSCs静脉移植可上调肺组织AQP1表达,加强肺泡液体清除能力,减轻肺损伤。

Abstract:

Objective To investigate the effect of the transplantation of human amniotic mesenchymal stem cell(hAMSCs)on the expressions of Aquaporin 1(AQP1)and phosphorylated p38 mitogen activated protein kinase(p-p38MAPK)in lung tissue of rats with acute lung injury(ALI), and to further explore whether it can alleviate lung injury and its possible mechanism. Methods 84 male SD rats were divided into 4 groups(n=21): normal saline control group(NS group), LPS induced ALI model group(LPS group), hAMSCs control group(NH group)and hAMSCs intervention group(LH group). The rats in LPS group and LH group were injected with LPS(4 mg/kg)through a sublingual vein to create models. NS group and NH group were injected with 250μl normal saline through a sublingual vein, and two hours later NH group and LH group were injected with 250μl DAP-labeled hAMSCs(2.5×10⁶)through a caudal vein. NS group and LPS group were injected with 250μl normal saline through a caudal vein. Seven rats in each group were killed at 6 h, 12 h and 72 h in order to observe the colonization of hAMSCs in the lung tissue of rats, the pathological morphology of lung tissue and the lung wet/dry weight(W/D)ratio. Furthermore, the levels of tumor necrosis factor-α(TNF-α)and interleukin- 1β(IL-1β)in bronchoalveolar lavage fluid(BALF)of rats in each group were detected by enzyme linked immunosorbent assay(ELISA). Immunohistochemistry(IHC)and western blot(WB)was used to detect the expressions of AQP1, p38 MAPK and p-p38 MAPK in the lung tissue of each group. Results ①Under the fluorescence microscope, it could be observed that there were blue fluorescent labeled hAMSCs around the alveoli of rats in NH and LH groups. Among them, the distribution of hAMSCs in LH group is more than that in NH group, indicating that hAMSCs can "homing" and colonize in the injured lung tissue. ②There was no significant difference in lung histopathological score, W/D ratio, TNF-α and IL-1 β levels in BALF, AQP1 and p-p38 MAPK protein expressions between NS group and NH group at each time point(P>0.05). ③Compared with the NS group and NH group at each time point, the LPS group and LH group showed an increase in lung histopathological score, W/D ratio, TNF-α and IL- 1 β levels in BALF, and p-p38 MAPK protein expression, and decrease in AQP1 expression at corresponding time points(P<0.05). ④Intra-group comparison of the LPS group revealed that there was no significant difference in lung histopathological score, W/D ratio, TNF-α and IL- 1 β levels in BALF, AQP1 and p-p38 MAPK protein expressions at the time points of 6 h and 12 h(P>0.05). Compared with LPS 6 h group and the LPS 12 h group, LPS 72 h group exhibited a decrease in lung histopathological score, W/D ratio, TNF-α and IL- 1 β levels in BALF, and p-p38 MAPK protein expression, and increase in AQP1 expression(P<0.05). Furthermore, there was no significant difference in lung histopathological score, W/D ratio, TNF-α and IL- 1 β levels in BALF, AQP1 and p-p38 MAPK protein expression between LH 6 h group and LH 12 h group(P>0.05). Compared with the 6 h and 12 h LH groups, LH 72 h group revealed a decrease in lung histopathological score, W/D ratio, TNF-α and IL- 1 β levels in BALF, and p-p38 MAPK protein expression, while the expression level of AQP1 was increased(P<0.05). Compared with the LPS group at each time point, lung histopathological score, W/D ratio, levels of TNF-α and IL-1β in BALF and expression of P-P38 MAPK protein were decreased in LH group, while AQP1 expression was increased(P<0.05). ⑤There were no significant difference in the expression of p38 MAPK in lung tissue at each time point among all groups(P>0.05). Conclusions ①There is a homing and colonization of exogenous hAMSCs in the injured lung tissue. ②hAMSCs can inhibit the activation of p38 MAPK, decrease the release of inflammatory factors TNF-α and IL- 1β, and alleviate pulmonary inflammatory response. ③Intravenous transplantation of hAMSCs can upregulate the expression of AQP1 in lung tissue to enhance alveolar fluid clearance, so as to alleviate lung injury.

参考文献/References:

1 马李杰, 李王平, 金发光. 急性肺损伤/急性呼吸窘迫综合征发病机制的研究进展[J/CD]. 中华肺部疾病杂志(电子版), 2013, 6(1): 65-68.
2 施 卉, 任成山. 急性肺损伤/急性呼吸窘迫综合征基础及临床研究进展[J/CD]. 中华肺部疾病杂志(电子版), 2013, 6(4): 350-355.
3 Liu C, Xiao K, Xie L. Advances in the use of exosomes for the treatment of ALI/ARDS[J]. Front Immunol, 2022, 13: 971189.
4 Derwall M, Martin L, Rossaint R. The acute respiratory distress syndrome: pathophysiology, current clinical practice, and emerging therapies[J]. Expert Rev Respir Med, 2018, 12(12): 1021-1029.
5 Long ME, Mallampalli RK, Horowitz JC. Pathogenesis of pneumonia and acute lung injury[J]. Clin Sci(Lond), 2022, 136(10): 747-769.
6 何 静, 郭 玲, 赵 庆, 等. 间充质干细胞治疗急性肺损伤的研究进展[J]. 中国临床医生杂志, 2022, 50(6): 651-654.
7 Anan HH, Zidan RA, Shaheen MA, et al. Therapeutic efficacy of bone marrow derived mesenchymal stromal cells versus losartan on adriamycin-induced renal cortical injury in adult albino rats[J]. Cytotherapy, 2016, 18(8): 970-984.
8 Su W, Cao R, Zhang XY, et al. Aquaporins in the kidney: physiology and pathophysiology[J]. Am J Physiol Renal Physiol, 2020, 318(1): F193-F203.
9 Tsushima K, King LS, Aggarwal NR, et al. Acute lung injury review[J]. Intern Med, 2009, 48(9): 621-630.
10 Liu L, Xie C. Effects of downregulation of aquaporin1 by peptidoglycan and lipopolysaccharide via MAPK pathways in MeT-5A cells[J]. Lung, 2011, 189(4): 331-340.
11 Li J, Xu M, Fan Q, et al. Tanshinone ⅡA ameliorates seawater exposure-induced lung injury by inhibiting aquaporins(AQP)1 and AQP5 expression in lung[J]. Respir Physiol Neurobiol, 2011, 176(1-2): 39-49.
12 Cui P, Xin H, Yao Y, et al. Human amnion-derived mesenchymal stem cells alleviate lung injury induced by white smoke inhalation in rats[J]. Stem Cell Res Ther, 2018, 9(1): 101.
13 Matute-Bello G, Downey G, Moore BB, et al. An official American Thoracic Society workshop report: features and measurements of experimental acute lung injury in animals[J]. Am J Respir Cell Mol Biol, 2011, 44(5): 725-738.
14 Palakshappa JA, Krall JTW, Belfield LT, et al. Long-term outcomes in acute respiratory distress syndrome: Epidemiology, mechanisms, and patient evaluation[J]. Crit Care Clin, 2021, 37(4): 895-911.
15 Xunian Z, Kalluri R. Biology and therapeutic potential of mesenchymal stem cell-derived exosomes[J]. Cancer Sci, 2020, 111(9): 3100-3110.
16 Lian J, Lin J, Zakaria N, et al. Acute lung injury: Disease modelling and the therapeutic potential of stem cells[J]. Adv Exp Med Biol, 2020, 1298: 149-166.
17 伍冬冬, 潘频华, 覃庆武. 急性肺损伤/急性呼吸窘迫综合征发病机制研究进展[J]. 中华结核和呼吸杂志, 2015, 38(7): 524-527.
18 Vassiliou AG, Manitsopoulos N, Kardara M, et al. Differential expression of aquaporins in experimental models of acute lung injury[J]. In Vivo, 2017, 31(5): 885-894.
19 沈慧萍, 麻海英, 洒玉萍, 等. 水通道蛋白1、5与肺水肿相关性研究进展[J]. 中国医学创新, 2021, 18(10): 181-184.
20 Wang T, Liu C, Pan LH, et al. Inhibition of p38 MAPK mitigates lung ischemia reperfusion injury by reducing blood-air barrier hyperpermeability[J]. Front Pharmacol, 2020, 11: 569251.
21 Liu L, Du L, Chen Y, et al. Down-regulation of Aquaporin1(AQP1) by peptidoglycan via p38 MAPK pathways in primary rat pleural mesothelial cells[J]. Exp Lung Res, 2014, 40(4): 145-153.
22 郭树明, 脱承德, 孟祥云. 宣白承气汤联合复合乳酸菌治疗脓毒症相关性ARDS的疗效分析[J]. 时珍国医国药, 2021, 32(1): 135-137.
23 Domscheit H, Hegeman MA, Carvalho N, et al. Molecular dynamics of lipopolysaccharide-induced lung injury in rodents[J]. Front Physiol, 2020, 11: 36.
24 Brooks D, Barr LC, Wiscombe S, et al. Human lipopolysaccharide models provide mechanistic and therapeutic insights into systemic and pulmonary inflammation[J]. Eur Respir J, 2020, 56(1): 1901298.
25 Nova Z, Skovierova H, Calkovska A. Alveolar-capillary membrane-related pulmonary cells as a target in endotoxin-induced acute lung injury[J]. Int J Mol Sci, 2019, 20(4): 831.
26 宣国平, 张 琳, 钟明媚. 脂多糖致大鼠急性肺损伤模型取材时间选择[J]. 中华实用诊断与治疗杂志, 2015, 29(2): 136-138.
27 王显峰, 白萨日娜. 脂多糖致小鼠急性肺损伤模型给药剂量和时间的筛选[J]. 中国兽医杂志, 2021, 57(1): 79-82+134.
28 Liang W, Guo L, Liu T, et al. MEF2C alleviates acute lung injury in cecal ligation and puncture(CLP)-induced sepsis rats by up-regulating AQP1[J]. Allergol Immunopathol(Madr), 2021, 49(5): 117-124.
29 Xu J, Huang B, Wang Y, et al. Emodin ameliorates acute lung injury induced by severe acute pancreatitis through the up-regulated expressions of AQP1 and AQP5 in lung[J]. Clin Exp Pharmacol Physiol, 2016, 43(11): 1071-1079.
30 Sun Y, Xia Y, Liu X, et al. Dexmedetomidine alleviates LPSinduced acute lung injury via regulation of the p38/HO1 pathway[J]. Mol Med Rep, 2020, 22(3): 2442-2450.
31 Wang WB, Li JT, Hui Y, et al. Combination of pseudoephedrine and emodin ameliorates LPS-induced acute lung injury by regulating macrophage M1/M2 polarization through the VIP/cAMP/PKA pathway[J]. Chin Med, 2022, 17(1): 19.
32 李榕生, 朱秀连, 邓春江, 等. 人羊膜间充质干细胞对松木屑烟雾诱导大鼠急性肺损伤的保护作用[J]. 华南国防医学杂志, 2022, 36(10): 765-768+777.

备注/Memo

备注/Memo:

基金项目: 黔科合支撑[2020]4Y162号
通信作者: 赵建军, Email: zhaojianjun1974@163.com

常用功能

更新日期/Last Update: 2023-04-20