Cigarette smoke extract combined with lipopolysaccharide reduces OCTN1/2 expression in human alveolar epithelial cells in vitro and rat lung in vivo under inflammatory conditions
Introduction
Over the last few decades, SLC22 transporters have been shown to play important roles in the absorption and distribution of drugs [1], [2]. The SLC22 family includes organic anion transporters (OATs), organic cation transporters (OCTs), and organic cation transporters novel (OCTNs) comprising over 400 genes [3], [4]. The OCTNs consist of OCTN1, OCTN2 and OCTN3, but OCTN3 is only found in mice and zebrafish [5]. Therefore, the OCTN subfamily is limited to OCTN1 and OCTN2, which can be used to explore the physiological and pharmacological effects. Related studies have shown that the expression of OAT1/3 is downregulated in the kidney tissue of mice with acute kidney injury, while the opposite is true with treatment [6]. The mRNA expression of OCT1/3 is downregulated in liver fibrosis mice, while the spontaneous reversal produces the opposite outcome [7]. The mRNA expression of OCT1, OCT2, OCT3 and OCTN1 is decreased in the kidney tissue of virus-infected rats [8]. Evidence is now emerging that decreased expression of OCTN2 leads to lack of carnitine, which increases the risk of inflammatory bowel disease, while variation in OCTN1 expression is related to rheumatoid arthritis and Crohn’s disease [9]. However, the change in OCTN1/2 expression under inflammatory conditions induced by CSE combined with LPS has not been explored in the lungs.
OCTN1/2 are highly expressed at the apical side of alveolar epithelial cells and in bronchial epithelial cells and transport organic cation, such as ergothioneine and carnitine [10]. OCTN1/2 are involved in the transport of drugs for the treatment of chronic pulmonary bronchitis, chronic obstructive pulmonary disease (COPD) and other pulmonary inflammatory diseases, such as IB, tiotropium bromide [11], [12], prednisolone [13]. Evidence has showed that the administration of L-carnitine can improve lung function in COPD patients [14], IB has been reported to be a common substrate of OCTN1/2 and is often used in the treatment of COPD as an anticholinergic drug [15]. The effects of drugs for COPD/inflammation on OCTN1/2 need to be further explored.
In this research, CSE combined with LPS was used to establish inflammation models in vitro and in vivo [16]. To examine the expression of OCTN1/2 in alveolar epithelial cells of the inflammation model and lung tissue in the rat COPD model induced by CSE combined with LPS, we analysed the effects of inflammation on OCTN1/2 expression. Through study of the effect of intervention with the COPD therapeutic drugs IB and DEX on OCTN1/2, we explored the potential impact of this effect on clinical medication and further explored the possible molecular mechanism.
Section snippets
Chemical reagents
Ipratropium Bromide (Item No. MB7513, Lot No. J1214AS) and dexamethasone (Item No. MB1434-S, Lot No. M0606AS) were purchased from Dalian Meilun Biotechnology Co., Ltd. LPS (Cat. No. L2880 E. coli 055:B5) and PDTC (Item No. P8765) were provided by Sigma (Biosciences, USA). Polyclonal rabbit anti-OCTN1 antibody (Item No. DF9724) was purchased from Affinity (Biosciences, USA). Polyclonal rabbit anti-OCTN2 antibody (Item No. bs-8149R) was supplied by Beijing Biosynthesis Biotechnology Co., Ltd.
H&E staining and immunohistochemistry
Using SD rats to establish a COPD model through CSE and LPS induction is a highly traditional and mature method. Histopathology analysis indicated that in the CSE-LPS model group, bronchiolar goblet cell numbers were increased, tracheal wall injury and remodeling alternately caused bronchoconstriction and airway obstruction, and alveolar rupture formed a large space. Compared with that in the control group, the alveolar diameter of the lung tissue in the CSE-LPS model group was significantly
Discussion
COPD is a serious public health problem that is characterized by persistent airflow limitation and a complex chronic inflammatory response [20], [21] caused by noxious gases or particles [22], [23]. Treatment of COPD has always been a difficult problem in the medical community [24], and drug transport and absorption are important factors for COPD treatment. The organic cation transporter OCTN1/2 paly an important role in drug transport [25]. In this study, we evaluated changes of in OCTN1/2 in
Conclusion
In conclusion, our study found that CSE combined with LPS could downregulate OCTN1/2 expression under inflammatory conditions and that the downregulation of OCTN1/2 expression might occur partially via the NF-κB signaling pathway. IB or DEX can upregulate OCTN1/2 expression under inflammatory conditions. These new findings provide new possibilities for curing COPD by increasing OCTN1/2 expression and regulating OCTNs-related signaling factors. These findings also have important clinical
Funding
This work was supported by the National Natural Science Foundation of China (grant number 81803868); Natural Science Foundation of Anhui Province (grant number 1308085QH153); Natural Science Foundation of Anhui Province (grant Number 1708085QH196); and Province Natural Science Foundation of Universities of Anhui Province (KJ2014A109).
Declaration of Competing Interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
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