Page 23 - ebook

Ginsenoside Rg1 protects polymorphonuclear
neutrophils from suppression of immune function by
mitochondrial damage-associated molecular patterns

Mina Boo , Hyo In Kim , Woo Yong Park , Gahee Song , Ja Yeon Park , Se Jin Jung , Minji Choi ,
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Jae-Young Um , Hyun Jeong Kwak , Jinbong Park ³,⁴*
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³,⁴
1 Department of Life Science, College of Natural Sciences, Kyonggi University, Suwon 16227, Korea
2 Department of Surgery, Beth Israel Deaconess Medical Center / Harvard Medical School, Boston 02215, USA
3 Department of Science in Korean Medicine, Graduate School, Kyung Hee University, Seoul 02447, Korea
4 Department of Pharmacology and Basic Research Laboratory for Comorbidity Research, College of Korean Medicine, Kyung Hee University,
Seoul 02447, Korea
Introduction Exposure to ND6 100 nM induced a notable decrease
in PMN respiratory burst in response to fMLF or LTB4.
Trauma is the leading cause of death in individuals However, Rg1 pre-treatment of 30 min revoked this
under 45 years old [1]. Trauma increases ND6-mediated respiratory burst suppression (Fig 1).
susceptibility to secondary infection and these
nosocomial infections are a common cause of
morbidity and mortality of trauma patients [2].
However, the biologic events linking injury to
suppression of anti-microbial immunity are not fully
understood. The injury-released danger associated
molecular patterns (DAMPs) modify polymorpho-
nuclear neutrophil (PMN) functions [3]. Ginsenoside
Rg1 (Rg1) is a saponin found in Panax ginseng, a
well-known anti-inflammatory herb [4]. We exposed Fig 1. fMLF-induced PMN respiratory burst.
healthy PMN with ND6, a mitochondrial DAMP, to Extracellular ROS is closely linked to NETs. Thus we
mimic a post-traumatic environment. We then studied examined the effect of Rg1 on NETosis. As seen in
the effects of Rg1 on the main PMN functions Fig 2, NETosis was suppressed by ND6, while Rg1
involved in microbial immunity. pre-treatment significantly revoked such suppression.
Materials and Methods Next, we assessed PMN chemotaxis after incubation
with ND6. As in Fig 3, ND6 significantly suppresses
PMN preparation: Fresh human PMN for functional PMN migration towards LTB4. However, pre-
assays were isolated from freshly withdrawn treatment with Rg1 reversed the ND6-induced
peripheral blood of healthy volunteers using 1-Step chemotaxis suppression (P < 0.01).
Polymorph (AN221725) gradient.
Respiratory burst assay: Reactive oxygen species
(ROS) production was measured by luminol-
dependent chemiluminescence in a 96-well plate
luminometer (Berthold) as previously described [5].
Neutrophil extracellular trap (NET) formation
assay: Phorbol 12-myristate 13-acetate (PMA)-
induced NETosis was assayed using the elastase
technique per manufacturer’s instructions (Item No.
601010, Cayman Chemical). Fig 2. PMA-induced PMN Fig 3. LTB4-induced PMN
Chemotaxis assay: PMN chemotaxis was studied in NETosis. chemotaxis.
3.0 μm-pore-transwells as described previously [5]. Discussion
Leukotriene B4 (LTB4) was used as the
chemoattractant. Our study demonstrates that Rg1 can enhance the
Statistical Analysis: Data were analyzed by analysis anti-microbial functions of PMN when subjected to
of variance (ANOVA) followed by Tukey’s post hoc mtDAMPs. The current study suggests a potential
test. P values <0.05 were considered statistically therapy for the critical unmet need of trauma patients.
significant. References
Results 1. WHO statistics (2020).
2. Sperry et al. N Engl J Med. 2018;379(4):315-26.
PMN migrate down chemical gradients towards areas 3. Itagaki et al. Crit Care Med. 2020;48(2):e123-e32.
of injury or infection by chemotaxis [6]. They can then 4. Lee and Lau. Molecules. 2011 Mar 30;16(4):2802-16.
form NETs that trap bacteria and use respiratory burst 5. Kim et al. J Trauma Acute Care Surg. Online ahead of print.
(ROS) to kill them [7,8]. We examined the change in 6. de Oliveira et al. Nat Rev Immunol. 2016;16(6):378-91.
these 3 events in PMN after ND6 exposure, and then 7. Jorgensen et al. Nat Rev Immunol. 2017;17(3):151-64.
evaluated the effect of Rg1. 8. Piacenza et al. J Exp Med. 2019;216(3):501-16.