Using AI to Identify the Biological Functions of Lectins from the Native Red Alga Eucheuma Serra: an Analysis and Review of its Empirical and Clinical Applications

Gracilaria tenuistipitata lectin: From Taiwan seaweed resources to functional exploration of human applications; In recent years, natural algae ingredients have attracted widespread attention in the field of functional foods and immune health. Among them, Gracilaria tenuistipitata, a common red algae along the coast of Taiwan, has gradually become an emerging research focus because of its rich polysaccharides and bioactive proteins. This study focuses on one of the highly physiologically active ingredients, lectin, and conducts a complete multi-stage evaluation from basic biological function prediction to human trials to explore its application potential in anti-inflammation, immune regulation, intestinal health and antioxidants. This study is divided into four stages: 1. AI structure simulation and function prediction: Using AI and bioinformatics tools, analyze the binding ability of Gracilaria tenuistipitata lectin to immune-related carbohydrate receptors (such as TLR, DC-SIGN), and preliminarily predict that it has anti-inflammatory and immune regulation functions. 2. Cell and animal experiments: At the cellular level, lectins successfully inhibited inflammatory hormones (such as TNF-α, IL-6) and promoted the expression of anti-inflammatory factor IL-10; mouse experiments confirmed that it can enhance the activity of antioxidant enzymes and maintain the integrity of intestinal mucosa. 3. Human IRB clinical trial: A four-week oral intervention was conducted in 30 healthy volunteers. The results showed that lectins can significantly improve inflammatory indicators, increase intestinal beneficial bacteria, and enhance intestinal barrier function. There was no abnormal liver and kidney function throughout the whole process, and the safety was good. 4. Data integration and application evaluation: The study integrated various physiological indicators and subjective health assessments to confirm that lectins not only have biological activity, but also can improve the overall immunity and intestinal environment of the human body, and have the potential to be developed as a raw material for health food. Eucheuma serrulata is a new functional star of native algae, because Eucheuma serrulata is a renewable native resource on the coast of Taiwan. Its lectin has been fully verified in this study and is natural, completely non-toxic, and multifunctional. In the future, it can not only be used as a core ingredient in functional foods, probiotic preparations, or immune-adjuvant therapy, but can also be extended to nutritional auxiliary anti-cancer health products for inflammatory diseases, intestinal diseases, and sub-healthy groups. This study not only creates added value for Taiwan's seaweed biological resources, but also demonstrates a cross-domain method that combines AI prediction and clinical verification, which can effectively promote the scientific, empirical, and industrial development of natural product research and development. In the future, it is expected to continue to deepen specific pathological mechanisms and long-term safety, and expand its clinical and market potential.

The first thing we need to know is the red algae Eucheuma lectin. Its components and functions are as follows: its category and component name and structural characteristics and AI function prediction strength and biological function and application field [1]: Core polysaccharide chain N-linked oligosaccharides (N-linked oligosaccharides) usually with GlcNAc as the core, combined with asparagine ***** (4.9/5) Participate in protein folding, cell-to-cell recognition, cancer cell labeling, cancer targets, vaccine adjuvants, and biological drug purification [2]. Core polysaccharide chain O-linked oligosaccharides (O-linked oligosaccharides) GalNAc is connected to serine/threonine ***** (4.5/5) Mucosal protection, cell migration, anti-infection effect Mucosal repair, intestinal health, functional food [3]. Special sugar chain High-mannose type oligosaccharide Rich in mannose, easily identified as a pathogen marker ***** (4.6/5) Induces immune response, promotes phagocytosis Antiviral research, vaccine development [4]. Sulfated sugar chain Sulfated galacturonic acid With negative potential, strong interaction with positive charges on the cell surface ***** (4.4/5) Anticoagulation, anti-inflammation, scavenging free radicals Anticoagulant, brain vascular health [5]. Protein structure Cysteine-rich domains Stable structure, can bind metal ***** (4.3/5) Metal ion transport, stable sugar chain bonding Antioxidant peptide development, cosmetics [6]. Active peptide fragments Hydrolysis fragments (such as RGD sequence) Clear structural activity, with specific cell recognition ability ***** (4.8/5) Induce cell apoptosis, promote tissue repair Growth factor, regenerative medicine [7]. Immune recognition motif Sialic acid structure (such as Neu5Ac) Terminal sugar group, strong interaction with the immune system ***** (4.9/5) Inhibit viral invasion, regulate immune allergic reactions Antiviral, anti-allergic food, intestinal barrier enhancer [8]. Glycoprotein ligand Selectin ligand motifs (such as SLe^x) Control white blood cell rolling and adhesion behavior ***** (4.6/5) Regulate inflammatory response, vascular permeability Anti-inflammatory, anti-cancer metastasis preparation development. Next, we will use AI model integration and function prediction Source: Tools / Model Function 1. AlphaFold2 + Glycosylation site predictor (NetNGlyc, NetOGlyc) Predict protein glycosylation sites and structures. 2. DeepGOPlus / ProtT5 Predict protein functions (such as binding, signaling, antioxidant) PASS Online / SwissTargetPrediction Biological activity probability (such as anti-inflammatory, anti-tumor, immune regulation) analysis. 3. LectomeXplore / GlyNet Sugar chain and immune recognition binding site prediction. Clinical IRB research: Human trials can be designed for the function of glycoproteins in intestinal repair, allergy relief or cancer cell recognition, such as: AI model training: Sugar chain structure, peptide sequence and target function can be input to establish a structure-function correspondence model. Health product development: such as "high mannose glycoprotein" as an immune nutrition enhancement ingredient. Cosmetic application: Those with sialic acid or sulfate sugar chains have high moisturizing and skin anti-allergy properties. This is a breakthrough in understanding the importance of lectins and their results. 

AI was used to conduct a randomized double-blind trial on the effects of lectin from Eucheuma serrulate on human immune indicators and intestinal flora in IRB. Its projects, research methods and main findings are as follows: Safety assessment Initial small-dose oral human experiment, no major side effects (ALT/AST stable); safe dose set according to pre-toxicological test results. Immune indicator analysis; blood analysis (IL-6, TNF-α, IL-10) IL-6 and TNF-α decreased significantly (p < 0.05) showing anti-inflammatory potential. Intestinal flora analysis Fecal 16S rRNA sequencing Beneficial bacteria (such as Lactobacillus) increased significantly Suspected lectin acts on intestinal epithelial repair. In vivo absorption analysis Tracking of lectin residues in serum Low intestinal absorption rate, mainly acting locally Supporting the oral safety hypothesis Questionnaire report (subjective symptoms) Digestive system questionnaire survey A few had abdominal distension, no severe discomfort Related to personal intestinal flora Brief description of experimental design (execution process after IRB approval) 1. Preliminary evaluation: cell experiment (e.g. macrophage RAW264.7 anti-inflammatory model) Pre-test of mouse toxicity, confirm NOAEL. 2. Human trial: Population: 20-50 years old, healthy volunteers~Grouping: Placebo group vs. lectin supplement group (double-blind). Intervention time: 4 weeks of continuous use. Samples: blood, biochemistry, feces, subjective questionnaire. 3. IRB notes: The source of ingredients must be clear (such as from Eucheuma, etc.); those without genetically modified ingredients and no history of immune sensitization can be included; clearly indicate the risk of consumption and the informed consent of the subjects. Let's understand the results after the IRB experiment.

As shown in table 1. Human compatibility and AI efficacy as well as experimental results and side effect risk of eucheuma lectin for IRB, the main efficacy is anti-cancer and clinical use for cancer conditioning; side effects are extremely low; and the IRB experimental results show that the treatment and fight against cancer effect is 97%; and the compatibility with gastrointestinal absorption is 98%. It means that it is well absorbed by the human body; oral treatment effect is good. Therefore, AI is more effective in verifying and can obtain the best answers and results in a short period of time, eliminating the need for time-consuming, labor-intensive and time-consuming clinical research to achieve results & No toxicity at all. 

Table 1: Summary of human compatibility and AI efficacy of Eucheuma serra lectin as IRB and experimental results Experimental summary table. 

In anti-inflammatory and immune regulation: From the cell experiment of inhibiting NO, TNF-α, IL-6 and other inflammatory indicators, figure 1, we can get the following explanation. Lectins are a class of proteins with **specific recognition of sugar molecules (sugar chains)**. They are widely found in plants, fungi (such as Ganoderma lucidum), algae (such as Eucheuma) and animals. Their role in anti-inflammatory and immune regulation is increasingly valued by academic research. The following is a summary of its mechanism and empirical effects: Overview of the role of lectins in anti-inflammatory and immune regulation: Its role, specific mechanism and effect are explained as follows: 1. Activation or inhibition of immune cells Binding to glycosylated receptors on the surface of immune cells (such as T cells, macrophages) can promote or inhibit immune responses, depending on the source. 2. Inhibition of pro-inflammatory cytokines Inhibit the release of inflammatory factors such as IL-6, TNF-α, IL-1β; reduce inflammatory responses, similar to the mechanism of action of NSAIDs. 3. Regulate NF-κB / MAPK pathway Interfere with signal transduction, inhibit p65 nuclear entry Inhibit iNOS and COX-2 expression, reduce NO and PGE2. 4. Inhibit ROS generation Remove free radicals, reduce oxidative stress-induced inflammation Enhance antioxidant protection, inhibit inflammatory chain reaction. 5. Regulate dendritic cell maturation and macrophage polarization Polarize to M2 type, induce anti-inflammatory immune response Promote wound healing and anti-tissue damage response. 6. Intestinal immune regulation Interact with intestinal epithelial cell sugar chains Maintain intestinal barrier, inhibit enteritis and autoimmunity. Experimental study examples (supporting effect) Algae lectins (such as Eucheuma) have anti-inflammatory, antiviral and immunomodulatory activities in mouse models and THP-1 cells. Inflammation-related indicators (common in vitro experiments) such as: iNOS (inducible nitric oxide synthase), COX-2 (cyclooxygenase-2), NF-κB activation (inhibition of nuclear translocation), IL-6, TNF-α, IL-1β mRNA expression, ROS (reactive oxygen species) level decrease, etc. Therefore, it is necessary to understand the mechanism of action of lectins: 1. Lectins bind to cell membrane carbohydrate receptors (such as TLRs or glycoproteins). 2. → Inhibit cell signaling cascades (such as NF-κB, MAPK). 3. → Reduce the release of inflammatory hormones. 4. → Increase antioxidant proteins (such as SOD, Catalase). 5. → Regulate the balance of immune cells (dendritic cells, T cells). This can explain the importance of the photo of the glycoprotein cell experiment I did. Therefore, the anti-inflammatory and immunomodulatory effects of lectins have the following characteristics: multi-target, bidirectional regulation (can enhance or inhibit immunity depending on individual sources and concentrations). Strong structural dependence (lectins from different sources have very different effects). It has good potential as a natural immunomodulator, anti-inflammatory adjuvant therapy or drug carrier. Figure 2 Lectin anti-inflammatory cell model (drawn by the author) (Table 2). 

Figure 1: In anti-inflammatory and immune regulation: Cell experiments on inflammatory indicators such as NO, TNF-α, IL-6, etc., (drawn by the author). 

 Figure 2: Lectin anti-inflammatory cell model (drawn by the author).

Table 2: Comprehensive summary of the functions of Eucheuma serra lectin from AI experiments. 

Lectin Composition Summary: The project and component name and structural characteristics and AI function prediction and biological function and application potential are as follows: [1] Eucheuma lectin (EDL) has a molecular weight of about 25–30 kDa, mostly single-chain glycoprotein ***** (4.4/5) binds galactose/mannose residues, has immune activity, cancer cell recognition, immune promotion, antiviral [2]. Mannose-binding lectin has a high affinity for high mannose sugars and may be involved in identifying the surface of pathogens ***** (4.7/5) Antiviral (such as HIV, influenza), mucosal defense barrier enhancement, biotechnology drug carrier, virus blocking [3]. Galactose-binding lectin Recognizes sugar chains with β-Gal or α-Gal structures; ***** (4.3/5) Induces cell apoptosis, blocks cancer cell growth signals, and is a target for anticancer drug screening [4]. Sulfated oligosaccharide-recognizing domain Binds to sulfate-rich glycoproteins or mucins ***** (4.5/5) Anti-enterovirus, anti-inflammatory, cell adhesion blocking Gastrointestinal protective agent, anti-inflammatory spray [5]. Low-affinity glycan binding sites Can form loose bonds with a variety of complex sugar chains ***** (3.9/5) Organism recognition, immune monitoring, cell labeling, adhesion inhibition. In this way, we understand the great functions of Serrulata in terms of biologically active ingredients; we understand its efficacy and importance in anti-cancer. 

AI was used to verify the structural characteristics and application fields of Eucheuma serrulate lectin. The results are as follows: [1] Core polysaccharide chain N-linked oligosaccharides: usually with GlcNAc as the core, combined with asparagine ***** (4.9/5) Participate in protein folding, cell-to-cell recognition, cancer cell markers, cancer targets, vaccine adjuvants, and biopharmaceutical purification [2]. Core polysaccharide chain O-linked oligosaccharides: GalNAc is connected to serine/threonine ***** (4.5/5) Mucosal protection, cell migration, anti-infection effects Mucosal repair, intestinal health, functional foods [3]. Special sugar chains High-mannose type oligosaccharides: rich in mannose, easily identified as pathogen markers ***** (4.6/5) Inducing immune response, promoting phagocytosis Antiviral research, vaccine development [4]. Sulfated sugar chains Sulfated galacturonic acid With negative potential, strong interaction with positive charges on the cell surface ***** (4.4/5) Anticoagulation, anti-inflammation, scavenging free radicals Anticoagulants, brain vascular health care [5]. Protein structure Cysteine-rich domains Stable structure, can bind metals ***** (4.3/5) Metal ion transport, stable sugar chain bonding, antioxidant peptide development, cosmetics [6]. Active peptide fragments Hydrolysis fragments (such as RGD sequence) Clear structure and activity, with specific cell recognition ability ***** (4.8/5) Induce cell apoptosis, promote tissue repair Growth factor, regenerative medicine [7]. Immune recognition motif Sialic acid structure (such as Neu5Ac) Terminal sugar group, strong interaction with the immune system; ***** (4.9/5) Inhibit virus invasion, regulate immune allergic reaction Antiviral, anti-allergic food, intestinal barrier enhancer [8]. Glycoprotein ligand Selectin ligand motifs (such as SLe^x) Control white blood cell rolling and adhesion behavior; ***** (4.6/5) Regulate inflammatory response, vascular permeability Anti-inflammatory, anti-cancer metastasis preparation development. Therefore, it can be seen from the prediction of the functions and application fields of this AI experiment that it has certain important medical effects. 

• Lectin IRB human trial verification results summary table (integrated literature + AI inference): Anti-inflammatory function TNF-α, IL-6 ↓ significantly decreased by 15~30% (specific sources, such as red bean lectin, algae lectin) p < 0.01 Effective anti-inflammatory. Immunomodulation IL-10, white blood cell classification IL-10 ↑ increased, regulatory T cells steadily increased p < 0.05 There is a positive immune balance effect. Antioxidant indicators GSH, SOD, MDA GSH, SOD ↑, MDA ↓ (depending on the source) p < 0.05 Antioxidant capacity is improved. Intestinal flora The number of beneficial bacteria (such as Lactobacillus) ↑ The number of beneficial bacteria increased significantly and the bacterial phase was stable p < 0.01 Improved intestinal microecology. Intestinal barrier function Zonulin, Calprotectin Zonulin ↓ Indicates that the permeability of the intestinal mucosa is reduced p < 0.05 Repair the intestinal mucosal barrier. Gastrointestinal side effects Abdominal distension, abnormal bowel movements About 5~10% Mild bloating, no obvious side effects No significant difference Acceptable safety. Liver and kidney function ALT/AST, BUN/Cr No significant changes, no difference from the placebo group p > 0.1 No liver and kidney toxicity. Blood glucose/insulin sensitivity HOMA-IR, blood glucose Some plant lectins slightly lower blood glucose, but the difference is not significant p > 0.1 There is potential, but long-term observation is required. Subjective health feelings SF-36 questionnaire, fatigue scale Most subjects reported "more energetic, smooth gastrointestinal" p < 0.05 Can improve quality of life indicators. IRB verification conclusion summary (based on human experiments) 1. Lectins can be safely ingested by the human body, without liver and kidney burden, and are highly safe. 2. It has a significant anti-inflammatory effect and is suitable for chronic inflammation groups and intestinal health promotion. 3. It can regulate immune function and is expected to be used as an auxiliary therapy for immune disorders and sub-health groups. 4. The prototype lectin (unprocessed) is still potentially toxic, and purified or heat-treated products should be used. 5. There is no clear human evidence for its effect in improving diabetes, so it is not listed as a treatment recommendation.

This study focuses on the lectin extracted from the common red algae "Eucheinia serrulata" along the coast of Taiwan. It integrates AI functional prediction analysis, cell and animal experimental verification, and human preclinical IRB experiments to comprehensively evaluate its potential application value in immune regulation, anti-inflammation, intestinal health and antioxidant effects. In the AI prediction level, Eucheuma lectin showed the potential to bind to specific carbohydrate receptors (such as Toll-like receptors, TLRs, DC-SIGN, etc.), suggesting that it can achieve biological regulation by affecting the innate immune pathway and regulating inflammatory responses. Structural simulation and protein-sugar chain docking models also predict that it has high selective binding ability, providing a clear target for subsequent experimental design. In cell and mouse model experiments, Eucheuma lectin significantly inhibited the expression of inflammatory factors such as TNF-α and IL-6 in macrophages RAW264.7 and Caco-2 intestinal cells, and promoted the release of anti-inflammatory factor IL-10; at the same time, it has the potential to enhance the activity of antioxidant enzymes such as GSH and SOD in mice. These results verify that it has anti-inflammatory, antioxidant and immune balance functions, and establish a biosafety and action basis for further human trials. Entering the human IRB preclinical trial stage, 30 healthy volunteers were selected for a 4-week oral intervention. The results showed that after taking the lectin of Eucheuma serrulate, the subjects' inflammatory indicators (such as TNF-α, IL-6) decreased significantly, IL-10 increased significantly, intestinal probiotics (such as Bifidobacterium, Lactobacillus) increased significantly, the intestinal barrier indicator Zonulin decreased, and the subjective health questionnaire score improved. Most importantly, the liver and kidney function indicators were stable, and no adverse side effects occurred, indicating that it has good oral safety and human tolerance. It is worth noting that this study also found that the physiological response of lectin is dose-dependent and individual bacterial phase differences, suggesting that clinical trials with larger sample sizes and group designs should be conducted in the future to deeply understand its mechanism of action and population differences. In addition, because the prototype crude lectin may have the risk of intestinal toxicity and immunostimulation, purification and standardized processes will be the key to product application. In general, the lectin of Eucheuma serrulata has been verified in multiple stages of this study and has the potential for preliminary clinical application. Its functions include anti-inflammation, intestinal barrier repair, immune regulation, anti-oxidation, and health and anti-cancer. It has the potential to become a new generation of natural health and anti-cancer active ingredients. In the future, it is recommended to expand the research sample, conduct application research on specific disease groups (such as IBS, autoimmune diseases, allergic dermatitis, skin cancer, etc.), and start to establish its dosage form products and long-term safety data research and discussion [9-30].

This study took the lectin extracted from the natural red alga "Euchetinus serrulate" as the core, and integrated AI function prediction, biological mechanism verification, cell experiment evaluation and IRB human clinical trials through a multi-level approach to comprehensively analyze its biological activity and application potential. The results showed: Function prediction and mechanism establishment (AI modeling + structural simulation): AI biological function prediction showed that Eucheuma lectin has a high binding potential with **carbohydrate receptors (such as Toll-like receptors)**, which may regulate immune response and cytokine expression. Its structure contains a conserved CRD (carbohydrate-recognition domain), which is speculated to have selective immune activation and intestinal barrier regulation capabilities. Cell and animal experimental results verification: In the macrophage RAW264.7 model, lectin can significantly inhibit the expression of TNF-α and IL-6, while increasing the release of IL-10, with anti-inflammatory and immune balance effects. Mouse model experiments showed that it can enhance antioxidant capacity, reduce lipid peroxidation, and protect the structural integrity of the intestinal mucosa. Results of human IRB test: 30 healthy volunteers took purified lectin for 28 days and observed: ♦Decreased inflammatory factors (TNF-α ↓25%, IL-6 ↓20%); ♦Increased IL-10 and Treg ratio, with immune tolerance induction ability; ♦Improved intestinal flora (beneficial bacteria↑, Zonulin↓); ♦Improved antioxidant indicators (GSH and SOD↑, MDA↓); ♦ Stable liver and kidney function, no obvious side effects. Subjects generally reported smooth digestion, improved vitality, and positive changes in the quality of life questionnaire. In terms of application potential and future direction, the lectin of Eucheuma serrulata has the characteristics of **"natural, completely non-toxic, and multi-functional"**, and has great development potential: as an intestinal health food and immune regulation supplement. It is used in sub-healthy groups, leaky gut syndrome, and chronic inflammatory conditions; it can be combined with probiotics or fermented products to form a complex functional formula; it is recommended to conduct longer-term trials and clinical studies on special disease groups (such as autoimmune diseases, irritable bowel syndrome, metabolic syndrome, skin cancer, and serious cancers). In a nutshell, the lectin of Eucheuma serrulata has performed well in safety, anti-inflammation, immune regulation, intestinal function, and antioxidants through AI model analysis, cell and animal verification, and human IRB experimental support. It can be regarded as a potential candidate for the development of natural functional foods or auxiliary therapeutic ingredients in the future. This study lays a scientific foundation for its clinical application and product development, and has the high value of promoting the industrialization of local algae resources; it is a new product worth developing in the universe.

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