LPAR1基因对脂多糖刺激下Ⅱ型肺泡上皮细胞中组织因子和纤溶酶原激活物抑制剂1表达的影响

doi:10.3969/j.issn.1006-5725.2023.18.007

Abstract

Abstract:

Objective To investigate the dynamic expression characteristics of lysophosphatidic acid receptor 1 （LPAR1）， a differentially expressed gene， in type Ⅱ alveolar epithelial cells （AEC Ⅱ） stimulated by lipopolysaccharide （LPS）， and the role of the gene in the regulation of tissue factor （TF） and plasminogen activator inhibitor-1 （PAI-1）. Methods RNA-seq sequencing was performed on RLE-6TN cells after injured by LPS， and then using bioinformatics analysis， identify the differentially expressed gene LPAR1 in the NF-κB signaling pathway. LPAR1 gene was dynamically expressed in RLE-6TN cells at 6， 12， 24， 48 and 72 h after LPS stimulation. On this basis， the expression of LPAR1 gene was knocked down by lentiviral transfection to observe the effect of the gene on the expression of TF and PAI-1. Results The expression of LPAR1 in RLE-6TN cells started to increase after 6 h of stimulation， peaked at 24 h， and then gradually decreased. LPAR1 gene levels were successfully knocked down through lentiviral transfection. After 24 hours of LPS stimulation， the expression levels of TF and PAI-1 mRNA and protein in the cells increased significantly compared with normal controls， with statistical significance （P < 0.05）， and then after knocking down LPAR1 gene， the expression levels of TF and PAI-1 decreased significantly （P < 0.05） and approached normal control values（P > 0.05）. Conclusion The LPAR1 gene has a regulatory effect on TF and PAI-1 in LPS-stimulated RLE-6TN cells. The gene is expected to be a new target for correcting fibrin deposition in the alveoli of ARDS.

Key words: lysophosphatidic acid receptor 1, type Ⅱ alveolar epithelial cells, acute respiratory distress syndrome, tissue factor, plasminogen activator inhibitor-1

CLC Number:

R441.8

Ying LIU,Xianjun CHEN,Chuan XIAO,Jia YUAN,Qing LI,Lu LI,Jinfeng YANG,Feng. SHEN. Regulatory effect of LPAR1 gene on tissue factor and plasminogen activator inhibitor-1 in LPSstimulated type II alveolar epithelial cells[J]. The Journal of Practical Medicine, 2023, 39(18): 2323-2329.

Figures/Tables 6

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References 26

1	MEYER N J， GATTINONI L， CALFEE C S. Acute respiratory distress syndrome［J］. Lancet， 2021， 398（10300）： 622-637.
2	KAKU S， NGUYEN C D， HTET N N，et al. Acute Respiratory Distress Syndrome： Etiology， Pathogenesis， and Summary on Management［Z］. J Intensive Care Med， 2020， 35（8）： 723-737.
3	MCCLINTOCK D， ZHUO H， WICKERSHAM N， et al. Biomarkers of inflammation， coagulation and fibrinolysis predict mortality in acute lung injury［J］. Crit Care， 2008， 12（2）： R41.
4	BOS L D J， WARE L B. Acute respiratory distress syndrome： causes， pathophysiology， and phenotypes［J］. Lancet， 2022. .
5	KOYAMA K， KATAYAMA S， TONAI K， et al. Biomarker profiles of coagulopathy and alveolar epithelial injury in acute respiratory distress syndrome with idiopathic/immune-related disease or common direct risk factors［J］. Crit Care， 2019， 23（1）： 283.
6	LIVINGSTONE S A， WILDI K S， DALTON H J， et al. Coagulation Dysfunction in Acute Respiratory Distress Syndrome and Its Potential Impact in Inflammatory Subphenotypes.［J］. Front Med （Lausanne）， 2021， 8： 723217.
7	BASTARACHE J A， WANG L， GEISER T， et al. The alveolar epithelium can initiate the extrinsic coagulation cascade through expression of tissue factor［J］. Thorax， 2007， 62（7）： 608-616.
8	BASTARACHE J A， FREMONT R D， KROPSKI J A， et al. Procoagulant alveolar microparticles in the lungs of patients with acute respiratory distress syndrome［J］. Am J Physiol Lung Cell Mol Physiol， 2009， 297（6）： L1035-1041.
9	刘雨睛，刘博，姚玲，等. 不同液体培养对脂多糖刺激后人Ⅱ型肺泡上皮细胞A549产生凝血及纤溶因子的影响［J］. 中国中西医结合急救杂志， 2016， 23（3）：294-298.
10	YANG G， LI X， LI Q， et al. Andrographolide Suppresses Expressions of Coagulation and Fibrinolytic Inhibition-Related Factors in LPS-Induced Alveolar Epithelial Cell Type II via NF-κB Signal Pathway In Vitro［J］. Intensive Care Res， 2022， 2（3-4）： 61-70.
11	李想，杨贵霞，沈锋，等. 黄连素剂量依赖性抑制脂多糖刺激下大鼠Ⅱ型肺泡上皮细胞促凝和纤溶抑制因子的表达［J］. 中华危重病急救医学， 2021， 33（1）：53-58.
12	郑兴昊，李清，沈锋，等. 大黄素对脂多糖刺激下肺泡Ⅱ型上皮细胞表达促凝及纤溶抑制因子的调节作用［J］. 实用医学杂志， 2022， 38（24）：3032-3036.
13	XIAO C, LI Q, XIAO J, et al. MiR-9 targeting RUNX1 improves LPS-induced alveolar hypercoagulation and fibrinolysis inhibition through NF-κB inactivation in ARDS［J］. Int Immunopharmacol, 2023, 120: 110318.
14	LIU B， WANG Y， WU Y， et al. IKKβ regulates the expression of coagulation and fibrinolysis factors through the NF-κB canonical pathway in LPS-stimulated alveolar epithelial cells type II［J］. Exp Ther Med， 2019， 18（4）： 2859-2866.
15	YANG H， QIAN H， LIU B， et al. Triptolide dose-dependently improves LPS-induced alveolar hypercoagulation and fibrinolysis inhibition through NF-κB inactivation in ARDS mice［J］. Biomed Pharmacother， 2021， 139： 111569.
16	CHENG Y， LIU B， QIAN H， et al. BAY11-7082 inhibits the expression of tissue factor and plasminogen activator inhibitor-1 in type-II alveolar epithelial cells following TNF-α stimulation via the NF-κB pathway［J］.Exp Ther Med， 2021， 21（2）： 177.
17	LIU B， WU Y， WANG Y， et al. NF-κB p65 Knock-down inhibits TF， PAI-1 and promotes activated protein C production in lipopolysaccharide-stimulated alveolar epithelial cells type II［J］. Exp Lung Res， 2018， 44（4-5）： 241-251.
18	LEE C H， SAPKOTA A， GAIRE B P， et al. NLRP3 Inflammasome Activation Is Involved in LPA（1）-Mediated Brain Injury after Transient Focal Cerebral Ischemia［J］. Int J Mol Sci， 2020， 21（22）：8595.
19	LEE J H， SARKER M K， CHOI H， et al. Lysophosphatidic acid receptor 1 inhibitor， AM095， attenuates diabetic nephropathy in mice by downregulation of TLR4/NF-κB signaling and NADPH oxidase［J］. Biochim Biophys Acta Mol Basis Dis， 2019， 1865（6）： 1332-1340.
20	GAIRE B P， SAPKOTA A， SONG M R， et al. Lysophosphatidic acid receptor 1 （LPA（1）） plays critical roles in microglial activation and brain damage after transient focal cerebral ischemia［J］. J Neuroinflammation， 2019， 16（1）： 170.
21	JIN Y， KIM E N， LIM J H， et al. Role of Aberrantly Activated Lysophosphatidic Acid Receptor 1 Signaling Mediated Inflammation in Renal Aging［J］. Cells， 2021， 10（10）：2580.
22	ZHAO J， HE D， SU Y， et al. Lysophosphatidic acid receptor 1 modulates lipopolysaccharide-induced inflammation in alveolar epithelial cells and murine lungs［J］. Am J Physiol Lung Cell Mol Physiol， 2011， 301（4）： L547-556.
23	OE Y， TAKAHASHI N. Tissue Factor， Thrombosis， and Chronic Kidney Disease［J］. Biomedicines， 2022， 10（11）：2737.
24	SUBRAMANIAM S， KOTHARI H， BOSMANN M. Tissue factor in COVID-19-associated coagulopathy［J］. Thromb Res， 2022， 220： 35-47.
25	GROVER S P， MACKMAN N. Tissue Factor： An Essential Mediator of Hemostasis and Trigger of Thrombosis［J］. Arterioscler Thromb Vasc Biol， 2018， 38（4）： 709-725.
26	BATIHA G E S， AL-KURAISHY H M， AL-MAIAHY T J， etal. Plasminogen activator inhibitor 1 and gestational diabetes： the causal relationship［J］. Diabetol Metab Syndr， 2022， 14（1）： 127.

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Regulatory effect of LPAR1 gene on tissue factor and plasminogen activator inhibitor-1 in LPSstimulated type II alveolar epithelial cells

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