Translational Medicine in Uremic Vascular Calcification: Scavenging ROS Attenuates p-Cresyl Sulfate-Activated Caspase-1, NLRP3 Inflammasome and Eicosanoid Inflammation in Human Arterial Smooth Muscle Cells

Jia-Feng Chang; Hsiao-Ling Kuo; Shih-Hao Liu; Chih-Yu Hsieh; Chih-Ping Hsu; Kuo-Chin Hung; Ting-Ming Wang; Chang-Chin Wu; Kuo-Cheng Lu; Wei-Ning Lin; Chi-Feng Hung; Wen-Chin Ko

doi:10.3390/life12050769

Translational Medicine in Uremic Vascular Calcification: Scavenging ROS Attenuates p-Cresyl Sulfate-Activated Caspase-1, NLRP3 Inflammasome and Eicosanoid Inflammation in Human Arterial Smooth Muscle Cells

Life (Basel). 2022 May 23;12(5):769. doi: 10.3390/life12050769.

Authors

Jia-Feng Chang^{1

2

3

4

5

6}, Hsiao-Ling Kuo⁷, Shih-Hao Liu⁸, Chih-Yu Hsieh^{1

4}, Chih-Ping Hsu^{9

10}, Kuo-Chin Hung⁶, Ting-Ming Wang¹¹, Chang-Chin Wu¹², Kuo-Cheng Lu¹³, Wei-Ning Lin¹⁴, Chi-Feng Hung¹⁵, Wen-Chin Ko^{15

16}

Affiliations

¹ Division of Nephrology, Department of Internal Medicine, En Chu Kong Hospital, New Taipei City 237, Taiwan.
² Division of Nephrology, Department of Internal Medicine, Taipei Medical University-Shuang Ho Hospital, New Taipei City 235, Taiwan.
³ Department of Nursing, Yuanpei University of Medical Technology, Hsinchu 300, Taiwan.
⁴ Renal Care Joint Foundation, New Taipei City 220, Taiwan.
⁵ Department of Internal Medicine, Armfulcare Hospital, Taoyuan City 320, Taiwan.
⁶ Division of Nephrology, Department of Medicine, Min-Sheng General Hospital, Taoyuan City 330, Taiwan.
⁷ Division of Rheumatology, Department of Internal Medicine, En Chu Kong Hospital, New Taipei City 237, Taiwan.
⁸ Division of Pathology, Department of Internal Medicine, En Chu Kong Hospital, New Taipei City 237, Taiwan.
⁹ Department of Biotechnology and Pharmaceutical Technology, Yuanpei University of Medical Technology, Hsinchu 300, Taiwan.
¹⁰ Department of Medical Laboratory Science and Biotechnology, Yuanpei University of Medical Technology, Hsinchu 300, Taiwan.
¹¹ Department of Orthopaedic Surgery, National Taiwan University Hospital, Taipei 100, Taiwan.
¹² Department of Orthopaedic Surgery, En Chu Kong Hospital, New Taipei City 237, Taiwan.
¹³ Division of Nephrology, Department of Medicine, Taipei Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, New Taipei City 231, Taiwan.
¹⁴ Graduate Institution of Biomedical and Pharmaceutical Science, College of Medicine, Fu Jen Catholic University, New Taipei City 242, Taiwan.
¹⁵ School of Medicine, Fu Jen Catholic University, New Taipei City 24205, Taiwan.
¹⁶ Division of Cardiac Electrophysiology, Department of Cardiovascular Center, Cathay General Hospital, Taipei 106, Taiwan.

Abstract

We formerly proved that uremic vascular calcification (UVC) correlates tightly with oxidative elastic lamina (EL) injury and two cell fates (apoptosis and osteocytic conversion) in smooth muscle cells (SMC) of chronic kidney disease (CKD) patients and eliminating p-cresyl sulfate (PCS)-activated intracellular ROS ameliorates the MAPK signaling pathway in a human arterial SMC (HASMC) model. Nonetheless, whether ROS scavenger attenuates PCS-triggered inflammasome activation and eicosanoid inflammation in the UVC process remains unknown. Patients with lower extremity amputation were categorized into CKD and normal control group according to renal function. We used immunohistochemistry stain to analyze UVC in arterial specimens, including oxidative injury (8-hydroxy-2'-deoxyguanosine (8-OHdG) and internal EL disruption), cytosolic phospholipase A2 (cPLA2), cyclooxygenase 2 (COX2), interleukin-1 beta (IL-1β), caspase-1 and NLRP3. To simulate the patho-mechanism of human UVC, the therapeutic effects of ROS scavenger on PCS-triggered inflammatory pathways was explored in a HASMC model. We found CKD patients had higher circulating levels of PCS and an increase in medial arterial calcification than the control group. In CKD arteries, the severity of UVC corresponded with expressions of oxidative EL disruption and 8-OHdG. Furthermore, coupling expressions of cPLA2 and COX2 were accentuated in CKD arteries, indicative of eicosanoid inflammation. Notably, tissue expressions of IL-1β, caspase-1 and NLRP3 were enhanced in parallel with UVC severity, indicative of inflammasome activation. From bedside to bench, ROS scavenger attenuates PCS-activated expressions of cPLA2/COX2, pro-caspase-1 and NLRP3 in the HASMC model. UVC as an inevitable outcome is predictive of death in CKD patients. Nonetheless, UVC remain pharmacoresistant despite the evolution of treatment for mineral-parathyroid hormone-vitamin D axis. Beyond the mineral dysregulation, the stimulation of pro-oxidant PCS alone results in eicosanoid inflammation and inflammasome activation. Concerning the key role of Caspase-1 in pyroptosis, cell fates of HASMC in uremic milieu are not limited to apoptosis and osteogenesis. In view of this, reducing ROS and PCS may act as a therapeutic strategy for UVC-related cardiovascular events in CKD patients.

Keywords: COX2; Caspase-1; IL-1β; NLRP3; cPLA2; inflammasome; p-cresyl sulfate; pyroptosis; reactive oxygen species; uremic vascular calcification.

Grants and funding

This work was financially supported by Ministry of Science and Technology (MOST 109-2628-B-385-001), the En Chu Kong Hospital (ECKH_W11005), and the Cathay General Hospital (CGH-MR-A11012). This research was also funded by the Renal Care Joint Foundation.