Effects of imperatorin derivatives on the activity and drug resistance protein of alveolar type Ⅱ epithelial cells in COPD

doi:10.11958/20240581

Abstract

Abstract:

Objective To explore effects of a derivative of imperatorin (IMP-1) on the activity and resistance proteins of alveolar type Ⅱ (ATⅡ) cells in chronic obstructive pulmonary disease (COPD). Methods AT Ⅱ cells were divided into the blank group (well growing AT Ⅱ cells cultured in conventional medium), the cigarette smoke extract (CSE) group (induced culture with 2% CSE added) and the IMP-1 group (intervention with 10 μ mol/L of IMP-1 on CSE induced AT Ⅱ cells). Proliferation rates of AT Ⅱ cells at 24, 48 and 72 hours were measured using MTT method. Apoptosis of ATⅡ cells was detected by Hoechst 33342 staining. The proportion of cells in each cycle was analyzed by flow cytometry. The number of ATⅡ cell clones was detected by soft AGAR clone formation assay. Multidrug resistance gene 1(MDR1) and multidrug resistance associated protein 1 (MRP1) were detected by Western blot assay. Results Compared with the blank group, the apoptosis rate and the proportion of G₀/G₁ phase of ATⅡ cell cycle were increased in the CSE group, and the proliferation rate and the proportion of S, G₂/M phase were decreased at 24 h, 48 h and 72 h (P<0.05). Compared with the CSE group, the cell proliferation rate, the proportion of S and G₂/M phases and the number of cell clones were increased in the IMP-1 group, and the proportion of G₀/G₁ phase in ATⅡ cell cycle and the protein levels of MDR1 and MRP1 were decreased in the IMP-1 group (P<0.05). Conclusion IMP-1 may downregulate the expression of MDR1 and MRP1 proteins induced by CSE in AT Ⅱ cells, inhibiting the progression of AT Ⅱ cells from S phase to G2 phase and from G2 phase to M phase, thereby inhibiting abnormal cell activation and increasing apoptosis.

Key words: pulmonary disease, chronic obstructive, imperatorin derivatives, alveolar type Ⅱ epithelial cells, cigarette smoke extract, multidrug resistance gene 1, multidrug resistance associated protein 1

CLC Number:

R563.9

Figures/Tables 7

References 20

[1]	侯万举, 曹洁, 王娟. 血清窖蛋白-1在慢性阻塞性肺疾病相关肺动脉高压患者中的表达及意义[J]. 天津医药, 2019, 47(2):155-158.
	HOU W J, CAO J, WANG J, et al. Expression and significance of serum cellarin-1 in patients with pulmonary hypertension associated with chronic obstructive pulmonary disease[J]. Tianjin Med J, 2019, 47(2):155-158. doi:10.11958/20181303.
[2]	MADDAH S A, BARZEGARI A. The effect of albuterol spray on hypoxia and bronchospasm in patients with chronic obstructive pulmonary disease (COPD) under general anesthesia:a bouble-blind randomized clinical trial[J]. Ethiopian J Health Sci, 2023, 33(3):491-498. doi:10.4314/ejhs.v33i3.12.
[3]	FIROOZI Z, SHAHI A, MOHAMMADISOLEIMANI E, et al. Circrna-associated cerna networks (circcenets) in chronic obstructive pulmonary disease (COPD)[J]. Life Sci, 2024,349:122715. doi:10.1016/j.lfs.2024.122715.
[4]	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. doi:10.3892/etm.2020.9608.
[5]	CLEMENTS D, MILLER S, BABAEI-JADIDI R, et al. Cross talk between LAM cells and fibroblasts may influence alveolar epithelial cell behavior in lymphangioleiomyomatosis[J]. Am J Physiol Lung Cell Mol Physiol, 2022, 322(2):L283-L293. doi:10.1152/ajplung.00351.2021.
[6]	李慧, 张吴霞, 熊烨. 欧前胡素调节MAPK/mTOR/p70S6K信号通路对急性髓系白血病细胞恶性生物学行为的影响[J]. 河北医药, 2023, 45(13):1941-1945.
	LI H, ZHANG W X, XUN Y. Influences of imperatorin on the malignant biological behaviors of acute myeloid leukemia cells by regulating the MAPK/mTOR/p70S6K signaling pathway[J]. Hebei Medical Journal, 2023, 45(13):1941-1945. doi:10.3969/j.issn.1002-7386.2023.13.004.
[7]	ZHENG X X, LIU J H, LIU M H, et al. Antidepressant effect and mechanism of imperatorin[J]. Molecules, 2022, 27(4):1179. doi:10.3390/molecules27041179.
[8]	NIKOLAOU V, MASSARO S, GARN W, et al. Fast decliner phenotype of chronic obstructive pulmonary disease (COPD):applying machine learning for predicting lung function loss[J]. BMJ Open Respir Res, 2021, 8(1):e000980. doi:10.1136/bmjresp-2021-000980.
[9]	KAHNERT K, JÖRRES RA, BEHR J, et al. The diagnosis and treatment of COPD and its comorbidities[J]. Dtsch Arztebl Int, 2023, 120(25):434-444. doi:10.3238/arztebl.m2023.027.
[10]	RABE K F, RENNARD S, MARTINEZ F J, et al. Targeting type 2 inflammation and epithelial alarmins in chronic obstructive pulmonary disease a biologics outlook[J]. Am J Respir Crit Care Med, 2023, 208(4):395-405. doi:10.1164/rccm.202303-0455CI.
[11]	于定荣, 翁小刚, 王本晓, 等. 延胡索、白芷炮制配伍对元胡止痛方中延胡索乙素、欧前胡素含量以及镇痛作用的影响[J]. 中草药, 2021, 52(8):2306-2312.
	YU D R, WENG X G, WANG B X, et al. Effect of processing and compatibility of Corydalis Rhizoma and Angelicae Dahuricae Radix on contents of tetrahydropalmatine and imperatorin in Yuanhu Zhitong Recipe and its analgesic effect[J]. Chinese Traditional and Herbal Drugs, 2021, 52(8):2306-2312. doi:10.7501/j.issn.0253-2670.2021.08.013.
[12]	王绍展, 朱青霞, 陈井霞, 等. 异欧前胡素通过NF-κB和JAK1/STAT1信号通路抑制巨噬细胞M1极化发挥抗炎镇痛作用[J]. 中南药学, 2023, 21(8):1985-1990.
	WANG S Z, ZHU Q X, CHENG J X, et al. Anti-inflammation and analgesic effect of isoimperatorin on the polarization of macrophage M1 via NF-κB and JAK1/STAT1 signaling pathway[J]. Central South Pharmacy, 2023, 21(8):1985-1990. doi:10.7539/j.issn.1672-2981.2023.08.002.
[13]	AMINI P, NODOOSHAN S J, ASHRAFIZADEH M, et al. Imperatorin attenuates the proliferation of MCF-7 cells in combination with radiotherapy or hyperthermia[J]. Curr Radiopharm, 2022, 15(3):236-241. doi:10.2174/1874471015666220318122202.
[14]	刘明娟, 韩瑞霞, 李铁, 等. 欧前胡素对人前列腺癌PC3细胞增殖凋亡及RhoA/ROCK1信号通路的影响[J]. 河北医学, 2021, 27(3):362-367.
	LIU M J, HAN R X, LI T. Effects of imperatorin on proliferation,apoptosis and RhoA/ROCK1 signaling pathway in human prostate cancer PC3 cells[J]. Hebei Medicine, 2021, 27(3):362-367.doi:10.3969/j.issn.1006-6233.2021.03.003.
[15]	王雅娴, 杨全军, 郭澄. 欧前胡素抗肿瘤作用机制及治疗的研究进展[J]. 中国医院药学杂志, 2022, 42(22):2425-2429.
	WANG Y X, YANG Q J, GUO C. Progress in antitumor research of imperatorin[J]. Chinese Journal of Hospital Pharmacy, 2022, 42(22):2425-2429. doi:10.13286/j.1001-5213.2022.22.20.
[16]	MAJA M, NENA M, JELENA R, et al. MDR1 gene polymorphisms and p-glycoprotein expression in respiratory diseases[J]. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub, 2015, 159(3):341-346. doi:10.5507/bp.2014.032.
[17]	TAGUCHI M, OYAMA R, KANESO M, et al. Hybrid QM/MM free-energy evaluation of drug-resistant mutational effect on the binding of an inhibitor indinavir to HIV-1 protease[J]. J Chem Inf Model, 2022, 62(5):1328-1344. doi:10.1021/acs.jcim.1c01193.
[18]	CHEN X Y, YANG Y, WANG J Q, et al. Overexpression of ABCC1 confers drug resistance to betulin[J]. Front Oncol, 2021, 11:640-656. doi:10.3389/fonc.2021.640656.
[19]	MÜHLFELD C, WREDE C, MOLNÁR V, et al. The plate body:3D ultrastructure of a facultative organelle of alveolar epithelial type Ⅱ cells involved in SP-A trafficking[J]. Histochem Cell Biol, 2021, 155(2):261-269. doi:10.1007/s00418-020-01912-7.
[20]	KRZYZANOWSKI D, KRUSZEWSKI M, GRZELAK A, et al. Differential action of silver nanoparticles on ABCB1(MDR1) and ABCC1(MRP1) activity in mammalian cell lines[J]. Materials(Basel), 2021, 14(12):3383. doi:10.3390/ma14123383.

组别	增殖率			凋亡率
组别	24 h	48 h	72 h	凋亡率
空白组	100.00±0.00	100.00±0.00	100.00±0.00	9.33±1.02
CSE组	35.45±1.08^a	41.46±1.21^a	50.33±1.35^a	85.57±2.34^a
IMP-1组	53.89±1.66^ab	69.76±1.84^ab	82.14±2.05^ab	40.26±1.59^ab
F	5 050.439^＊＊	3 195.432^＊＊	1 893.672^＊＊	2 925.154^＊＊

组别	增殖率			凋亡率
组别	24 h	48 h	72 h	凋亡率
空白组	100.00±0.00	100.00±0.00	100.00±0.00	9.33±1.02
CSE组	35.45±1.08^a	41.46±1.21^a	50.33±1.35^a	85.57±2.34^a
IMP-1组	53.89±1.66^ab	69.76±1.84^ab	82.14±2.05^ab	40.26±1.59^ab
F	5 050.439^＊＊	3 195.432^＊＊	1 893.672^＊＊	2 925.154^＊＊

组别	G₀/G₁/%	S/%	G₂/M/%	克隆数目/个
空白组	8.31±0.33	46.10±5.56	45.59±5.49	148.33±3.36
CSE组	53.85±5.85^a	22.62±0.83^a	23.53±0.78^a	78.57±1.15^a
IMP-1组	21.78±2.49^ab	39.56±3.10^ab	38.66±3.65^ab	115.13±2.61^ab
F	242.836^＊＊	64.096^＊＊	52.028^＊＊	48.120^＊＊

组别	G₀/G₁/%	S/%	G₂/M/%	克隆数目/个
空白组	8.31±0.33	46.10±5.56	45.59±5.49	148.33±3.36
CSE组	53.85±5.85^a	22.62±0.83^a	23.53±0.78^a	78.57±1.15^a
IMP-1组	21.78±2.49^ab	39.56±3.10^ab	38.66±3.65^ab	115.13±2.61^ab
F	242.836^＊＊	64.096^＊＊	52.028^＊＊	48.120^＊＊

组别	MDR1	MRP1
空白组	1.02±0.04	1.10±0.02
CSE组	2.35±0.15^ab	2.51±0.13^a
IMP-1组	1.60±0.09^ab	1.57±0.07^ab
F	229.663^＊＊	450.337^＊＊