Recombinant thrombomodulin may protect cardiac capillary endothelial glycocalyx through promoting Glypican-1 expression under experimental endotoxemia

Yoshinori Kakino; Tomoaki Doi; Hideshi Okada; Kodai Suzuki; Chihiro Takada; Hiroyuki Tomita; Hirotaka Asano; Soichiro Kano; Yugo Wakayama; Tomoki Okuda; Hirotsugu Fukuda; Ayane Nishio; Yuki Kawasaki; Ayumi Kuroda; Takuto Shimada; Shigeo Takashima; Keiko Suzuki; Genki Yoshimura; Ryo Kamidani; Ryu Yasuda; Tetsuya Fukuta; Yuichiro Kitagawa; Haruka Okamoto; Takahito Miyake; Akio Suzuki; Takahiro Yoshida; Nobuyuki Tetsuka; Shozo Yoshida; Shinji Ogura

doi:10.1016/j.heliyon.2022.e11262

Recombinant thrombomodulin may protect cardiac capillary endothelial glycocalyx through promoting Glypican-1 expression under experimental endotoxemia

Heliyon. 2022 Oct 25;8(11):e11262. doi: 10.1016/j.heliyon.2022.e11262. eCollection 2022 Nov.

Authors

Yoshinori Kakino¹, Tomoaki Doi¹, Hideshi Okada¹, Kodai Suzuki¹, Chihiro Takada¹, Hiroyuki Tomita², Hirotaka Asano¹, Soichiro Kano¹, Yugo Wakayama¹, Tomoki Okuda¹, Hirotsugu Fukuda¹, Ayane Nishio¹, Yuki Kawasaki¹, Ayumi Kuroda¹, Takuto Shimada¹, Shigeo Takashima³, Keiko Suzuki^{4

5}, Genki Yoshimura¹, Ryo Kamidani¹, Ryu Yasuda¹, Tetsuya Fukuta¹, Yuichiro Kitagawa¹, Haruka Okamoto¹, Takahito Miyake¹, Akio Suzuki⁴, Takahiro Yoshida¹, Nobuyuki Tetsuka⁵, Shozo Yoshida^{1

6}, Shinji Ogura¹

Affiliations

¹ Department of Emergency and Disaster Medicine, Gifu University Graduate School of Medicine, Gifu, Japan.
² Department of Tumor Pathology, Gifu University Graduate School of Medicine, Gifu, Japan.
³ Division of Genomics Research, Life Science Research Center, Gifu University, Gifu, Japan.
⁴ Department of Pharmacy, Gifu University Hospital, Gifu, Japan.
⁵ Department of Infection Control, Gifu University Graduate School of Medicine, Gifu, Japan.
⁶ Abuse Prevention Center, Gifu University Graduate School of Medicine, Gifu, Japan.

Abstract

Introduction: Myocardial dysfunction occurs in patients with sepsis due to vascular endothelial injury. Recombinant human thrombomodulin (rhTM) attenuates vascular endothelial injuries through endothelial glycocalyx (eGC) protection.

Hypothesis: We hypothesized that rhTM attenuates myocardial dysfunction via the inhibition of vascular endothelial injury during sepsis.

Methods: Ten-week-old male C57BL6 mice were injected intraperitoneally with 20 mg/kg of lipopolysaccharide (LPS). In rhTM-treated mice, rhTM was injected intraperitoneally at 3 and 24 h after LPS injection. Saline was injected intraperitoneally as control. To assess for eGC injury, intensity score was measured 48 h after the LPS injection. To confirm vascular endothelial injuries, ultrastructural analysis was performed using scanning (SEM) and transmission electron microscopy (TEM).

Results: The survival rate of the rhTM group at 48 h after LPS injection was significantly higher than that of the control group (68% vs. 17%, p < 0.05). The serum level of troponin I in the rhTM group was lower than that in the control (2.2 ± 0.4 ng/dL vs 9.4 ± 1.1 ng/dL, p < 0.05). The expression of interleukin-6 (IL-6) was attenuated in the rhTM-treated group than in the control (65.3 ± 15.3 ng/mL vs 226.3 ± 19.4 ng/mL, p < 0.05). The serum concentration of syndecan-1, a marker of glycocalyx damage, was significantly decreased 48 h post-administration of LPS in the rhTM-treated group than in the control group. In ultrastructural analysis using SEM and TEM, eGC peeled off from the surface of the capillary lumen in the control. Conversely, the eGC injury was attenuated in the rhTM group. Gene set enrichment analysis revealed that osteomodulin, osteoglycin proline/arginine-rich end leucine-rich repeat protein, and glypican-1, which are proteoglycans, were preserved by rhTM treatment. Their protein expression was retained in endothelial cells.

Conclusion: rhTM attenuates sepsis-induced myocardial dysfunction via eGC protection.

Keywords: Glycocalyx; Glypican-1; Microvascular dysfunction; Thrombomodulin; Vascular endothelium.