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Article

Sarcodia suieae Acetyl-Xylogalactan Regulates Nile Tilapia (Oreochromis niloticus) Tissue Phagocytotic Activity and Serum Indices

by
Po-Kai Pan
1,†,
Tsung-Meng Wu
1,†,
Chiu-Ming Wen
2,
Yin-Yu Chen
3 and
Yu-Sheng Wu
1,*
1
Department of Aquaculture, National Pingtung University of Science and Technology, Pingtung 912301, Taiwan
2
Department of Life Sciences, National University of Kaohsiung, Kaohsiung 811726, Taiwan
3
Department of Aquaculture, National Taiwan Ocean University, Keelung 202301, Taiwan
*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this work.
J. Mar. Sci. Eng. 2022, 10(1), 18; https://doi.org/10.3390/jmse10010018
Submission received: 22 November 2021 / Revised: 19 December 2021 / Accepted: 21 December 2021 / Published: 24 December 2021
(This article belongs to the Special Issue Nutrition and Immunity for Sustainable Marine Aquaculture Development)
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Abstract

:
Sarcodia suieae acetyl-xylogalactan was reported to induce macrophage polarisation, and could positively regulate macrophage activation. In this study, we evaluated the effect of Sarcodia suieae acetyl-xylogalactan on the Nile tilapia. First, we assessed the influence of acetyl-xylogalactan on the survival, glucose uptake, and phagocytic activity of tilapia head kidney (THK) melanomacrophage, and observed increased proliferation of these cells in the MTT assay after 12 and 24 h of treatment. Glucose uptake increased in THK melanomacrophage treated with 20 and 30 μg acetyl-xylogalactan for 24 h. Their phagocytic activity was positively enhanced following exposure to acetyl-xylogalactan. Nile tilapia were fed with acetyl-xylogalactan for 4 weeks. At the end of the experiment, Nile tilapia were sacrificed, and the lipopolysaccharide-induced liver and head-kidney apoptosis was examined under reducing conditions in comparison with controls. The phagocytic activities of liver and head-kidney cells were enhanced after 4 weeks of feeding. Blood biochemical analysis revealed a reduction in glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) levels after 4 weeks of feeding. Combined with in vitro and in vivo experiments results, the extracted S. suieae acetyl-xylogalactan could directly induce THK melanomacrophage proliferation, glucose uptake, and phagocytic activity. Acetyl-xylogalactan was able to induce Nile tilapia liver and head-kidney resident macrophage activity, and reduced LPS-induced liver and head-kidney cell apoptosis. S. suieae acetyl-xylogalactan may modulate Nile tilapia macrophage activation by polarising them into M1 macrophages to improve the Nile tilapia nonspecific immune response.

1. Introduction

Polysaccharides from algae exhibit immunoregulatory properties [1]. Algae have been used for thousands of years. Marine macroalgae, and red and green algae contain high levels of proteins, polysaccharides, long-chain fatty acids, and other biocompounds [2,3]. Polysaccharides play an important role in the energy storage and the basal structure of marine algae [2,3]. To our knowledge, the underlying mechanisms of the functions of most immunoregulatory substances have been studied in mouse models but not in aquatic animal models.
Macrophages from animal tissue and organs that respond to chemokines are exposed to antigens as pathogens [4]. Dunaliella [5], Haematococcus [6], Scenedesmus [7], and Sarcodia [8] contain functional materials that have been studied in many animal models. These bioactive substances exert immunoregulatory functions in response to infections and immune diseases. Sea alga polysaccharides could bind to dectin-1 and activate macrophages, neutrophils, and dendritic cells [8]. Macrophages are primarily divided into two types on the basis of their functions and differentiation abilities, namely, classically and alternatively activated macrophages, which play a role in inflammation and wound healing, respectively [9].
In our previous study, the polysaccharide acetyl-xylogalactan extracted from sea alga Sarcodia suieae was analysed for its mono-/polysaccharide content, monosaccharide composition, acetyl content, and molecular weight by matrix-assisted laser desorption/ionisation-time of flight (MALDI-TOF) mass spectrometry and nuclear magnetic resonance (NMR) [10]. Acetyl-xylogalactan could directly induce tumour necrosis factor (TNF), interleukin (IL)-1, and mucosa-associated lymphoid tissue lymphoma translocation protein 1 (Malt1) production, and downregulate IL-6 and IL-17A expression, thereby regulating the inflammatory response via the nuclear factor kappa B (NF-κB) pathway [10].
In the present study, we examined the bioactivity of xylogalactan in the Nile tilapia. First, we used the tilapia head-kidney (THK) melanomacrophage cell line as the target in the presence or absence of acetyl-xylogalactan. THK macrophage survival, glucose uptake, and phagocytic activity were assessed in the in vitro study. According to a previous study, haematological parameters are valuable indicators of fish health status [11,12,13]. Moreover, haematological parameters are a valuable approach for monitoring the effects of changes in fish physiology [14,15]. We also investigated whether acetyl-xylogalactan functions in the physiological response of the Nile tilapia. In the in vivo experiment, Nile tilapia were fed with acetyl-xylogalactan for 4 weeks to detect lipopolysaccharide (LPS)-induced apoptosis, phagocytosis, and blood biochemical parameters.

2. Results

2.1. In Vitro Activation of THK Cells

2.1.1. Proliferation Effect of Acetyl-Xylogalactan on THK Cells

THK melanomacrophage were used as the experimental model and treated with different concentrations (0, 10, 20, and 30 μg/mL) of acetyl-xylogalactan. Their survival was analysed on the basis of OD values. Results showed that the activation of succinate dehydrogenase was similar between acetyl-xylogalactan-treated and untreated THK cells at 12 and 24 h (Figure 1A,B). At 12 h, the 20 and 30 μg treatment groups showed a significant induction of succinate dehydrogenase as compared to the control group (p < 0.01, Figure 1A). At 24 h, the level of succinate dehydrogenase was significantly higher in the 30 μg treatment group than that in the control group (p < 0.05, Figure 1B).
Thus, acetyl-xylogalactan induced THK cell proliferation, as evident from the result of the MTT assay.

2.1.2. Induction of Intracellular Glucose Uptake in THK Cells

The effect of acetyl-xylogalactan on the intracellular glucose uptake of THK cells was examined using a commercial kit. Results presented in Figure 2A,B show THK cells treated with 10, 20, and 30 μg acetyl-xylogalactan. At 12 h, there was no significant difference between the treatment and control groups (p > 0.05, Figure 2A). At 24 h, however, 20 and 30 μg acetyl-xylogalactan treatment significantly increased the intracellular uptake of glucose in THK cells (p < 0.05, Figure 2B). Therefore, acetyl-xylogalactan induced THK macrophage glucose uptake.

2.1.3. Effect of Acetyl-Xylogalactan on the Phagocytotic Activity of THK Cells

We measured the phagocytosis of E. coli by THK cells. Results presented in Figure 3A,B show that 10, 20, and 30 μg/mL acetyl-xylogalactan treatment significantly induced the phagocytosis ability of THK cells (p < 0.05) at 24 h (Figure 3B). However, acetyl-xylogalactan did not significantly enhance THK macrophage phagocytic activation at 12 h (p > 0.05).
Phagocytosis assessment is used as an important observation of macrophage activation parameters. During this experiment, acetyl-xylogalactan induced THK phagocytosis activity.

2.2. In Vivo Experiment with Nile Tilapia

2.2.1. Effect of Acetyl-Xylogalactan on LPS-Induced Apoptosis

LPS-induced apoptosis of cells is an important event in the regulation of inflammation.
The effect of acetyl-xylogalactan on LPS-induced liver apoptosis was examined. As shown in Figure 4A, treatment with acetyl-xylogalactan reduced liver cell apoptosis induced by LPS. After LPS treatment for 12 h, cells treated with 10% acetyl-xylogalactan showed a significant decrease in apoptosis (p < 0.05, Figure 4A, right panel). After 36 h of LPS treatment, the 10% and 15% treatment groups showed a significant reduction in apoptosis as compared to the control group (p < 0.05, Figure 4A, left panel).
Analysis of the head-kidney cells revealed no significant difference in LPS-induced apoptosis following treatment with acetyl-xylogalactan at 12 h (p > 0.05, Figure 4B, right panel). However, the rate of apoptosis significantly decreased in the group exposed to LPS for 36 h and fed with 5%, 10% and 15% acetyl-xylogalactan (p < 0.01, Figure 4B, left panel). Thus, acetyl-xylogalactan could reduce the LPS-induced apoptosis of cells, as evident from the Annexin V staining result.

2.2.2. Effect of Acetyl-Xylogalactan on Phagocytotic Activity

Phagocytosis is an important process in the regulation of immune activation. The effect of acetyl-xylogalactan on liver-resident macrophages was examined. Treatment with acetyl-xylogalactan induced the phagocytotic activity of liver-resident macrophages (Figure 5A). Feeding test animals with 15% acetyl-xylogalactan led to a significant increase in the phagocytic activity of macrophages (p < 0.05).
We evaluated the effect on head-kidney macrophages and found that 10% acetyl-xylogalactan treatment significantly affected the phagocytotic activity of head kidney macrophages (p < 0.05, Figure 5). Taken together, acetyl-xylogalactan treatment at various concentrations for 4 weeks induced the phagocytic activity of liver and head-kidney macrophages.

2.2.3. Blood Biochemical Analysis

Haematologic and blood biochemical parameters reflect alterations in physiological responses. During 4 weeks of acetyl-xylogalactan feeding, the effects on total cholesterol (T-Cho), blood urea nitrogen (BUN), total bilirubin (T-Bil), glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT), total protein (T-pro), albumin (Alb), calcium (Ca), and uric acid (UA) were analysed.
Feeding with 5%, 10%, and 15% acetyl-xylogalactan increased glucose levels at week 4 (p < 0.01) as compared to the control treatment (Table 1, column presentation). Total cholesterol level was significantly higher in the 10% feeding group than that in the control group (p < 0.05), but significantly decreased in the 15% treatment group (p < 0.05) at week 4 (Table 1). A comparison between the first and fourth week analyses revealed that feeding with 5% and 15% acetyl-xylogalactan led to a significant decrease in total cholesterol level (p < 0.05), as shown in Table 1 (row presentation). GOT and GPT analyses showed that feeding with 10% and 15% acetyl-xylogalactan significantly reduced these parameters (p < 0.05, Table 1, column presentation). A comparison between the first- and fourth-week analyses revealed a significant decrease in GOT and GPT levels after 10% and 15% acetyl-xylogalactan treatment (p < 0.01, Table 1, row presentation). Total protein level decreased in the group fed with 15% acetyl-xylogalactan for 4 weeks as compared to that in the control group (p < 0.05, Table 1, column presentation). Intracellular calcium was significantly increased in the control, and 5% and 15% feeding groups on the 4th week observation as compared to the 1st week (p < 0.05, Table 1, row presentation).
No significant differences were observed in the BUN, total bilirubin, albumin, and UA levels between different groups (p > 0.05).

3. Discussion

In this study, the functional effects of S. suieae acetyl-xylogalactan were investigated using THK cell and Nile tilapia experiments. THK cells were treated with various concentrations of S. suieae acetyl-xylogalactan, and the cellular response was analysed in terms of survival, phagocytic activity, and glucose uptake. In addition, Nile tilapia were fed with different concentrations of acetyl-xylogalactan, and the phagocytic ability of liver and head-kidney macrophages was assessed. Liver and kidney macrophages were sampled at the end of the treatment and treated with LPS to analyse the apoptotic response by examining changes in peripheral-blood parameters. Our findings revealed that S. suieae acetyl-xylogalactan positively enhanced the phagocytic activity and reduced the LPS-induced apoptosis of these cells. Biochemical analyses demonstrated the protective effects of S. suieae acetyl-xylogalactan on Nile tilapia, as evident from changes in the levels of GOT, GPT, and other haematologic factors.
Studies have revealed the signalling mechanisms involved in inflammation regulation. S. suieae acetyl-xylogalactan activates intracellular NF-κB signalling, and promotes the production of cytokines such as IL-1, IL-6, IL-8, and TNF. These effects lead to the activation of the M1 macrophage phenotype [8]. M1 macrophages play a major role in proinflammatory cytokine production and function against infection [16,17]. A previous study showed that Lycium barbarum polysaccharide could promote inflammatory cytokine production by activating p38 mitogen-activated protein kinase (MAPK) phosphorylation and reducing c-Junction N-terminal kinase (JNK) and extracellular signal-regulated kinase 1/2 (ERK1/2) MAPK phosphorylation [18,19]. Here, S. suieae acetyl-xylogalactan induced macrophage activation in Nile tilapia, consistent with observations reported in mouse models.
M1 macrophages preferentially metabolise glucose as an energy substrate, which plays a role in the activation of inflammatory responses by classically activated M1 macrophages [20]. The proinflammatory immune response of M1 macrophages enhances the expression of glucose transporter isoform 1 (Glut1), glucose-6-phosphate dehydrogenase, and hexokinase, which increase the macrophage glucose uptake response [21]. Glucose metabolism is a complicated intracellular phenomenon involving several different regulators. S. suieae acetyl-xylogalactan stimulation may activate NF-κB in Nile tilapia, thereby promoting Glut1 expression and inducing glucose uptake.
Evidence suggests that LPS enhances C/EBP homologous protein (CHOP), an endoplasmic reticulum (ER) stress-induced transcription factor that regulates the apoptosis response [22]. A previous study in an LPS-stimulated BALB/c mouse model investigated the effect of pretreatment with Astragalus polysaccharides (APS) on intestinal inflammation [23]. APS reduced the production of inflammatory factors and chemokines, and inhibited the LPS-induced activation of MAPK and NF-κB inflammatory pathways. Investigation of the polyphenol-rich compound extracted from Ecklonia cava showed a reduction in LPS-induced inflammation in a zebrafish model and demonstrated the protective effect against LPS-induced toxicity on zebrafish embryos [24]. Acorus tatarinowii polysaccharide was also shown to exert protective functions and prevent LPS-induced neurotoxicity by inhibiting Toll-like receptor 4 (TLR4)-mediated MyD88/NF-κB and phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signalling pathways [25]. In our study, feeding Nile tilapia with various concentrations of S. suieae acetyl-xylogalactan for 4 weeks lead to the amelioration of LPS-induced liver and head-kidney cell apoptosis. In particular, the head kidney is the most important immune organ in the Nile tilapia. Excessive inflammatory response or apoptosis of the head kidney may affect the immune response during infection. S. suieae acetyl-xylogalactan may protect against bacterial or endotoxin-induced injury.
GOT and GPT are the most important parameters for assessing fish health status [26,27]. In this study, Nile tilapia fed with various concentrations of S. suieae acetyl-xylogalactan had lower levels of GOT and GPT than those of the control group. This observation is indicative of the decreased stress level after acetyl-xylogalactan feeding. In addition, BUN level reveals the status of the kidney and liver [28]. Increased BUN level is indicative of acute kidney injury and associated with increased oxidative stress and free radical-induced damage [29,30]. In our study, feeding Nile tilapia with acetyl-xylogalactan for 4 weeks did not increase BUN levels as compared to other groups. Total bilirubin level reflects the metabolic process of bile components. Hepatocytes intake indirect bilirubin, combine it with Y and Z proteins [31], and then transport it to the ER. In the ER, indirect bilirubin binds to glucuronic acid to form bilirubin [32]. Total bilirubin level is used to evaluate liver function, as it is released into the blood during liver damage. As shown in our present study, acetyl-xylogalactan did not induce any increase in total bilirubin level. Analyses of GOT, GPT, BUN, and total bilirubin showed that acetyl-xylogalactan did not injure the liver. However, total protein, albumin, calcium, and UA levels were not significantly changed after acetyl-xylogalactan treatment.
This research illustrated that the extracted S. suieae acetyl-xylogalactan could directly induce THK melanomacrophage proliferation, glucose uptake, and phagocytic activity. Acetyl-xylogalactan was able to induce the Nile tilapia liver and head-kidney resident macrophage activity, and reduced LPS-induced liver and head-kidney cell apoptosis. S. suieae acetyl-xylogalactan potentially modulates Nile tilapia macrophage activation by polarising them into M1 macrophages.

4. Materials and Methods

4.1. S. suieae Acetyl-Xylogalactan and Feed Preparation

The preparation method of S. suieae acetyl-xylogalactan reported in our previous study [27] was used. Briefly, dried S. suieae was extracted in deionisation and distilled water at 60 °C for 6 h. The polysaccharide was then extracted by ethanol and ice-dried in a refrigerator at 4 °C. The experimental 10, 20, and 30 μg/mL dosage and treatment time followed the previous study [10].
This research was divided into an in vitro and an in vivo experiment. First, S. suieae acetyl-xylogalactan was treated with the THK cell to observe THK cell proliferation, glucose uptake, and phagocytotic activity. Then, feeding the Nile tilapia with various concentrations of the S. suieae acetyl-xylogalactan for 4 weeks detected LPS-induced apoptosis, tissue (liver and head kidney) phagocytotic activity, and serum indices. The research design is shown in Figure 6.

4.2. In Vitro Experiments

4.2.1. THK Melanomacrophage Cell Line

The THK melanomacrophage cell line was obtained from Prof. Chiu-Ming Wen (Department of Life Sciences, National University of Kaohsiung). Cells were maintained in 90% L-15 medium supplemented with 10% foetal bovine serum and 1% antibiotics (antibiotic antimycotic solution 100× liquid w/10,000 U penicillin, 10 mg streptomycin, and 25 µg amphotericin B per millilitre in 0.9% normal saline) and incubated at 27 °C without CO2. The THK cell was cultured in a 175 T flask contained and incubated at 27 °C. Waiting for 4–5 days of culture, the cultured THK cell was removed by the cell scraper, and cell numbers were adjusted as 1 × 106 cells by cell-counting machine (Scepter ™, Millipore, Merck KGaA, Darmstadt, Germany).

4.2.2. Proliferation Effect of S. suieae Acetyl-Xylogalactan on THK Cells

To examine the toxicity of S. suieae acetyl-xylogalactan on THK cells, the experimental groups (1 × 106 cells) were treated with or without acetyl-xylogalactan powder at 10, 20, and 30 μg/mL concentrations for 12 and 24 h. After incubation, cells were treated with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay (M2128, Sigma, Merck KGaA, Darmstadt, Germany) reagent. THK cells were plated in 96-well plates, giving a final volume of 200 μL of culture medium. Cells were adjusted to 106 cells per well and incubated at 27 °C for 24 h. At the end of incubation, the cultured medium was removed, and 10 μL (5 mg/mL) of MTT (SIGMA) was loaded with the medium and incubated at 27 °C for 4 h. Thereafter, 200 μL of dimethylsulfoxide (DMSO) was added to dissolve the formazan, and absorbance at 570 nm was measured using a microplate reader.

4.2.3. Glucose Uptake of THK Cells Treated with S. suieae Acetyl-Xylogalactan

To examine whether S. suieae acetyl-xylogalactan affected cellular glucose uptake, the experimental THK (1 × 106 cells) were treated with or without 10, 20, and 30 μg/mL S. suieae acetyl-xylogalactan for 12 and 24 h. After incubation, cells were subjected to Glucose Uptake Colorimetric Assay (K676-100, BioVision Inc., Milpitas, CA, USA), and absorbance was measured at 412 nm wavelength using a microplate reader.

4.2.4. Effect of S. suieae Acetyl-Xylogalactan on the Phagocytosis of THK Cells

The phagocytotic activity of THK cells was determined by pHrodo Green BioParticles® Conjugates for Phagocytosis (P35366, Thermo Fisher Scientific Inc., Waltham, MA, USA). Briefly, 1 × 106 THK cells were cultured in the medium at 27 °C without 5% CO2 for 24 h, and then treated with different concentrations (10, 20, and 30 μg/mL) of S. suieae acetyl-xylogalactan for 12 and 24 h. The culture medium was removed after treatment, and cells were incubated with bioparticles for 2 h. Phagocytosis activity was detected at Ex/Em = 509/533 using a fluorescence microplate reader.

4.3. In Vivo Experiment

The diet was prepared according to the commercial tilapia feed (SHYE YIH FEEDING Co., Ltd., Qieding, Kaohsiung, Taiwan). Dried acetyl-xylogalactan was dissolved in sterile D.D. water and sprayed onto the commercial feed. The food was gently mixed at room temperature (25 °C) and baked at 60 °C for complete drying.
Experimental feed containing 0%, 5%, 10%, or 15% acetyl-xylogalactan was used to feed Nile tilapia twice daily; the daily feed intake of Nile tilapia was equal to 5% of their weight at 8:00 a.m. and 16:00 p.m. Siphon cleaning of the tank bottom was conducted every 2 days. The feeding experiment was performed for 4 weeks. At the end of the experiment, tilapias were sacrificed, and their liver, head kidney, and peripheral blood were collected.
Tilapia (Oreochomis niloticus) fish were obtained from the Sheng-Diao Aquatic Technology, Pingtung, Taiwan. After transferring to the laboratory, tilapias were cultured until the body weight of each tilapia was approximately 30 g. This experiment used 5 individuals per group and experiment was replicated for 3 times. In total, 60 fish (Nile tilapia) were used in this research.

4.3.1. LPS-Induced Liver and Head Kidney Cell Apoptosis

To examine whether S. suieae acetyl-xylogalactan protected the Nile tilapia against LPS-induced apoptosis, the liver and head kidney were sampled from the groups treated without (control) or with (5%, 10%, and 15%) acetyl-xylogalactan. Three fish were selected from each experimental tank.
The liver and head-kidney samples were ground using a cell strainer in L-15 medium. The target liver and head-kidney cells were collected by 70 μm pore size cell strainer, and their density was adjusted to 1 × 106 cells/well. Prepared liver and head-kidney cells were treated with or without 100 μg/mL LPS for 12 and 36 h at 27 °C without 5% CO2. At the end of the treatment period, Annexin V-FITC Apoptosis Detection kit (AVK250, Strong Biotech Corporation, Nankang, Taipei, Taiwan) was used to measure the degree of apoptosis at excitation (Ex) and emission (Em) wavelengths of 488 and 530 nm, respectively.

4.3.2. Phagocytosis Activity of Liver and Head-Kidney Cells

To examine whether S. suieae acetyl-xylogalactan protected the Nile tilapia against LPS-induced apoptosis, liver and head-kidney samples were obtained following treatment without (control) or with (5%, 10%, and 15%). After 4 weeks of feeding, three fish from each group were sacrificed, and their liver and head-kidney samples were ground using a cell strainer in L-15 medium. In brief, the liver and head kidney were centrifuged at 4000× rpm for 30 min with 30% and 50% Percoll [33]. The layer between 30% and 50% Percoll was collected after centrifugation. Resident liver and head-kidney macrophages were used in this study. The target cell density was adjusted to 1 × 106 cells/well, and cells were incubated for 24 h. The culture medium was removed, and cells were treated with bioparticles for 2 h. Phagocytosis activity was detected at Ex/Em = 509/533 using a fluorescence microplate reader.

4.3.3. Blood Biochemical Analysis

Three fish were obtained from each experimental tank at the end of the treatment. Blood samples (1 mL) were collected from the caudal vessels using heparinised syringes, and placed in sterile tubes for biochemical analysis. Any haemolysed, clotted, or insufficient samples were discarded. Plasma samples were analysed for glucose, total cholesterol, blood urea nitrogen (BUN), total bilirubin, aspartate aminotransferase, alanine aminotransferase, total protein, albumin, calcium, and uric acid (UA) using an automated blood biochemical analyser (SPOTCHEM EZ SP-4430, ARKRAY Inc., Kamigyō-ku, Kyoto Japan).

4.4. Statistical Analysis

Data displayed differences between the treatment and control groups. We used the t-test, Scheffe’s test, and one-way analysis of variance (ANOVA) to analyse statistical significance between the treatment and control groups. Statistical significance was set at p < 0.05. Results are presented as mean ± standard deviation (SD; * p < 0.05, ** p < 0.001).

5. Conclusions

In summary, the extracted S. suieae acetyl-xylogalactan could directly induce THK melanomacrophage proliferation, glucose uptake, and phagocytic activity. In the feeding experiment, S. suieae acetyl-xylogalactan induced Nile tilapia liver and head-kidney resident macrophage activity, and reduced LPS-induced liver and head-kidney cell apoptosis. Thus, S. suieae acetyl-xylogalactan potentially modulates Nile tilapia macrophage activation by polarising them into M1 macrophages. Analysis of haematological indices showed that feeding Nile tilapia with S. suieae acetyl-xylogalactan had no negative effect.

Author Contributions

P.-K.P. and T.-M.W.: data curation and formal analysis; Y.-Y.C. and C.-M.W.: sample analysis and data curation; Y.-S.W.: data curation, formal analysis and article writing. All authors have read and agreed to the published version of the manuscript.

Funding

Council of Agriculture, Executive Yuan, Taiwan, ROC (no. 109-12.3.1-a4).

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Acknowledgments

Thanks to the National Pingtung University of Science and Technology and National Taiwan Ocean University for supporting the project.

Conflicts of Interest

The authors declare no conflict of interest.

Ethical Declarations

Experiment fish care and handling procedures in the present study were approved by the Laboratory Animal Center, National Pingtung University of Science and Technology.

Abbreviations

Total cholesterol (T-Cho), blood urea nitrogen (BUN), total bilirubin (T-Bil), glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT), total protein (T-pro), albumin (Alb), calcium (Ca), and uric acid (UA)).

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Jmse 10 00018 g001a 550Jmse 10 00018 g001b 550
Figure 1. Proliferation effect of acetyl-xylogalactan on THK macrophages. (A) THK cells treated with acetyl-xylogalactan for 12 h; (B) THK cells treated with acetyl-xylogalactan for 24 h. * p < 0.05; ** p < 0.01. All treatment groups were compared to the control group.
Figure 1. Proliferation effect of acetyl-xylogalactan on THK macrophages. (A) THK cells treated with acetyl-xylogalactan for 12 h; (B) THK cells treated with acetyl-xylogalactan for 24 h. * p < 0.05; ** p < 0.01. All treatment groups were compared to the control group.
Jmse 10 00018 g001aJmse 10 00018 g001b
Jmse 10 00018 g002a 550Jmse 10 00018 g002b 550
Figure 2. Glucose uptake effect of acetyl-xylogalactan on THK macrophages. (A) THK cells treated with acetyl-xylogalactan for 12 h; (B) THK cells treated with acetyl-xylogalactan for 24 h. * p < 0.05. All treatment groups were compared to the control group.
Figure 2. Glucose uptake effect of acetyl-xylogalactan on THK macrophages. (A) THK cells treated with acetyl-xylogalactan for 12 h; (B) THK cells treated with acetyl-xylogalactan for 24 h. * p < 0.05. All treatment groups were compared to the control group.
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Jmse 10 00018 g003a 550Jmse 10 00018 g003b 550
Figure 3. Effects of acetyl-xylogalactan on phagocytotic activity of THK macrophages. (A) THK cells treated with acetyl-xylogalactan for 12 h; (B) THK cells treated with acetyl-xylogalactan for 24 h. * p < 0.05. All treatment groups were compared to the control group.
Figure 3. Effects of acetyl-xylogalactan on phagocytotic activity of THK macrophages. (A) THK cells treated with acetyl-xylogalactan for 12 h; (B) THK cells treated with acetyl-xylogalactan for 24 h. * p < 0.05. All treatment groups were compared to the control group.
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Figure 4. Effect of acetyl-xylogalactan on LPS-induced apoptosis. (A) Liver of Nile tilapia fed with acetyl-xylogalactan for 4 weeks was treated with LPS. (B) Head kidney of Nile tilapia fed with acetyl-xylogalactan for 4 weeks was treated with LPS. * p < 0.05; ** p < 0.01. All treatment groups were compared to the control group.
Figure 4. Effect of acetyl-xylogalactan on LPS-induced apoptosis. (A) Liver of Nile tilapia fed with acetyl-xylogalactan for 4 weeks was treated with LPS. (B) Head kidney of Nile tilapia fed with acetyl-xylogalactan for 4 weeks was treated with LPS. * p < 0.05; ** p < 0.01. All treatment groups were compared to the control group.
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Figure 5. Effect of acetyl-xylogalactan on phagocytotic activities of Nile tilapia liver and head-kidney cells. (A) Phagocytotic activity of liver cells of Nile tilapia fed with acetyl-xylogalactan for 4 weeks. (B) Phagocytotic activity of head-kidney cells of Nile tilapia fed with acetyl-xylogalactan for 4 weeks. * p < 0.05. All treatment groups were compared to the control group.
Figure 5. Effect of acetyl-xylogalactan on phagocytotic activities of Nile tilapia liver and head-kidney cells. (A) Phagocytotic activity of liver cells of Nile tilapia fed with acetyl-xylogalactan for 4 weeks. (B) Phagocytotic activity of head-kidney cells of Nile tilapia fed with acetyl-xylogalactan for 4 weeks. * p < 0.05. All treatment groups were compared to the control group.
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Figure 6. Experimental design graph. (A) In vitro design; (B) in vivo experiment design.
Figure 6. Experimental design graph. (A) In vitro design; (B) in vivo experiment design.
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Table
Table 1. Serum indices of Nile tilapia fed with acetyl-xylogalactan for 4 weeks.
Table 1. Serum indices of Nile tilapia fed with acetyl-xylogalactan for 4 weeks.
1st Week4th Weekp Value
T-ChoControl125 ± 4.35128.33 ± 2.3040.306
5%133.66 ± 2.30 *127.66 ± 1.520.019
10%142.33 ± 1.52 *138.66 ± 2.51 *0.097
15%139.33 ± 2.08 *116.33 ± 1.52 *0.0001
BUNControl<5<5-
5%<5<5-
10%<5<5-
15%<5<5-
T-BilControl<0.2<0.2-
5%<0.2<0.2-
10%<0.2<0.2-
15%<0.2<0.2-
GOTControl114.33 ± 5.1361.66 ± 2.08<0.0001
5%109 ± 7.5456.33 ± 1.52 *<0.0001
10%112.33 ± 6.5027.66 ± 1.52 **<0.0001
15%143.33 ± 3.21 *24.66 ± 0.57 **<0.0001
GPTControl61.33 ± 2.0844 ± 4.350.0034
5%51.66 ± 2.0826.66 ± 0.570.0016
10%67.66 ± 0.5711.66 ± 0.57 **<0.0001
15%61 ± 1.7318 ± 1.73 *<0.0001
T-ProControl2.4 ± 02.6 ± 0.050.007
5%2.6 ± 0.12.7 ± 0.10.205
10%3.3 ± 0.6 *2.6 ± 0.10.129
15%2.5 ± 0.052.4 ± 0*0.007
AlbControl<0.1<0.1-
5%<0.1<0.1-
10%<0.1<0.1-
15%<0.1<0.1-
CaControl17.1 ± 0.518.56 ± 0.550.026
5%15.4 ± 0.8819.46 ± 0.50.023
10%17.4 ± 0.9818.63 ± 0.550.1313
15%17.53 ± 0.4118.76 ± 0.610.0446
UAControl<1<1-
5%<1<1-
10%<1<1-
15%<1<1-
Note: * p < 0.05; ** p < 0.01 (column presentation). Each treatment group was compared to the control group at the same week. p value (row presentation) indicates the comparison between the first and fourth week for each group.
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Pan, P.-K.; Wu, T.-M.; Wen, C.-M.; Chen, Y.-Y.; Wu, Y.-S. Sarcodia suieae Acetyl-Xylogalactan Regulates Nile Tilapia (Oreochromis niloticus) Tissue Phagocytotic Activity and Serum Indices. J. Mar. Sci. Eng. 2022, 10, 18. https://doi.org/10.3390/jmse10010018

AMA Style

Pan P-K, Wu T-M, Wen C-M, Chen Y-Y, Wu Y-S. Sarcodia suieae Acetyl-Xylogalactan Regulates Nile Tilapia (Oreochromis niloticus) Tissue Phagocytotic Activity and Serum Indices. Journal of Marine Science and Engineering. 2022; 10(1):18. https://doi.org/10.3390/jmse10010018

Chicago/Turabian Style

Pan, Po-Kai, Tsung-Meng Wu, Chiu-Ming Wen, Yin-Yu Chen, and Yu-Sheng Wu. 2022. "Sarcodia suieae Acetyl-Xylogalactan Regulates Nile Tilapia (Oreochromis niloticus) Tissue Phagocytotic Activity and Serum Indices" Journal of Marine Science and Engineering 10, no. 1: 18. https://doi.org/10.3390/jmse10010018

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