This protocol describes a retrospective cohort method to evaluate combined saxagliptin and liraglutide therapy on inflammation and adipokine profiles in obese type 2 diabetes patients with poor response to metformin.
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Method Article
This protocol describes a retrospective cohort method to evaluate combined saxagliptin and liraglutide therapy on inflammation and adipokine profiles in obese type 2 diabetes patients with poor response to metformin.
Metformin is the first-line therapy for obese patients with type 2 diabetes mellitus (T2DM); however, a substantial proportion of patients exhibit inadequate glycemic control despite optimal dosing. This protocol describes a retrospective cohort approach to evaluate the effects of saxagliptin combined with liraglutide on microinflammation and adipokine profiles in obese T2DM patients with poor response to metformin. A total of 116 eligible patients were identified from electronic medical records and assigned to receive either saxagliptin monotherapy or saxagliptin combined with liraglutide based on predefined clinical criteria. The protocol includes standardized procedures for patient selection, group allocation, metabolic assessment, and laboratory measurement of inflammatory markers, adipokines, and metabolic parameters.
Representative outcomes demonstrate that both treatment regimens are associated with improvements in glycemic control, lipid metabolism, and insulin resistance. The combination regimen shows greater reductions in inflammatory markers and more favorable modulation of adipokine profiles compared to monotherapy, while maintaining a comparable safety profile. These findings illustrate the application of this protocol for evaluating multi-target metabolic interventions in real-world clinical settings.
This method provides a reproducible framework for assessing combined pharmacological strategies in obese T2DM patients with suboptimal response to first-line therapy and may support further clinical and translational investigations.
Key words: Saxagliptin; Liraglutide; Obesity-related type 2 diabetes; inflammatory response; Microinflammatory response
T2DM is a global epidemic, with its incidence increasing year by year and posing a major public health threat. Epidemiological surveys indicate approximately 463 million diabetes patients worldwide, over 90% of whom have T2DM. Obesity is one of the most important risk factors for T2DM1. In China, rapid economic development and lifestyle changes have led to a sharp rise in obesity‑related T2DM, creating a “dual disease burden”. These patients often present not only with significant glucose metabolism disorders but also with lipid abnormalities, insulin resistance (IR), chronic low‑grade inflammation, and dysregulated adip....
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1. Ethical Approval
2. Patient Identification and Eligibility Screening
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A total of 116 obese T2DM patients with poor metformin response (HbA1c ≥ 7.5% despite metformin ≥ 1500 mg·day-1 for ≥ 12 weeks) were retrospectively included (Table 1). Patients were assigned to a control group (saxagliptin alone, n = 58) or a combination group (saxagliptin plus liraglutide, n = 58). Baseline demographic and clinical characteristics were comparable between the two groups (all p > 0.05). The following sections present representative results for metabolic, inflammato.......
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With the development of society and the change in dietary patterns, the clinical characteristics of diabetes patients have shown new trends. Among T2DM patients, the traditional “three more and one less” symptoms are no longer common. Data indicate that 44% of T2DM patients did not exhibit typical clinical symptoms, and overweight/obesity has a high prevalence in the T2DM population, often associated with insulin resistance, lipid metabolism disorders, and chronic hyperglycemia32. Metf.......
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Conflicts of Interest
The authors declare that they have no financial conflicts of interest.
This work was supported by Lu Hao Huangpu District Famous Doctor Studio of General Practice & Speciality (Grant No.2023QZ03).
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| Name | Company | Catalog Number | Comments |
|---|---|---|---|
| 32G×4 mm disposable needles | BD Medical | 328418 | For subcutaneous injection of liraglutide; 4 mm length reduces intramuscular injection risk |
| 5810R | Refrigerated centrifuge | Eppendorf | 5810R |
| Adiponectin ELISA kit | R&D Systems | DY1065 | For quantitative measurement of human adiponectin in serum; 96-well plate format |
| AU680 | Fully automated biochemical analyzer | Beckman Coulter | AU680 |
| Bluetooth heart rate monitor | Xiaomi | Mi Band 6 | For monitoring exercise heart rate; 1 Hz sampling frequency; syncs with research tablet |
| Chemerin ELISA kit | R&D Systems | DY2325 | For detection of human chemerin (RARRES2) in serum; sensitivity <50 pg/mL |
| Cobas e601 | Fully automated chemiluminescence immunoanalyzer | Roche Diagnostics | Cobas e601 |
| C-reactive protein (CRP) ELISA kit | Abcam | ab260078 | For high-sensitivity measurement of human CRP; range 0.1–100 ng/mL |
| Date-labeled medication boxes | MedCenter | M0314 | Daily pill organizer with 7 compartments; helps track medication adherence |
| Dietary Analysis and Meal Planning System | Dietary planning software | Nutritionist Pro | Version 2.0 or higher |
| EDTA anticoagulant tubes (purple cap) | BD Vacutainer | 367861 | For HbA1c and complete blood count; 5 mL draw volume; spray-coated EDTA |
| Electronic weight scale | Tanita | HD-351 | For body weight measurement; capacity 150 kg, accuracy 0.1 kg |
| Exercise log (paper) | In-house | N/A | Custom-designed form to record exercise type, duration, heart rate, and RPE score |
| Fluoride/oxalate anticoagulant tubes (gray cap) | BD Vacutainer | 367835 | For glucose measurement; inhibits glycolysis; 3 mL draw volume |
| Fully automated biochemical analyzer | Beckman Coulter | AU680 | For measurement of FPG, 2hPG, TC, TG, HDL-C, LDL-C; throughput up to 800 tests/h |
| Fully automated chemiluminescence immunoanalyzer | Roche Diagnostics | Cobas e601 | For fasting insulin (FINS) measurement; electrochemiluminescence method |
| G*Power | Heinrich-Heine-Universität Düsseldorf, Germany | RRID:SCR_013726 | Free statistical power analysis software. The version used in this study is 3.1.9.6. Official download and information page: https://www.psychologie.hhu.de/arbeitsgruppen/allgemeine-psychologie-und-arbeitspsychologie/gpower. Key reference: Faul, F., Erdfelder, E., Lang, A.-G., & Buchner, A. (2007). G*Power 3: A flexible statistical power analysis program for the social, behavioral, and biomedical sciences. Methods of Behavioral Research, 39(2), 175-191. |
| G11 | HbA1c analyzer (HPLC) | Tosoh Bioscience | G11 |
| HbA1c analyzer (HPLC) | Tosoh Bioscience | G11 | For glycated hemoglobin quantification; NGSP-certified; retention time 2.4 min |
| IL-6 ELISA kit | R&D Systems | DY206 | For human IL-6 detection in serum; sensitivity <0.7 pg/mL |
| Liraglutide pre-filled injection pen | Novo Nordisk | J20160005 | Pre-filled 3 mL (18 mg) pen for subcutaneous injection; delivers 0.6, 1.2, or 1.8 mg/dose |
| Medication log (paper) | In-house | N/A | Daily record for drug intake, adverse events, and injection site rotation |
| Mi Band 6 | Wearable heart rate monitor | Xiaomi | Mi Band 6 |
| Microplate reader (ELISA) | BioTek | Synergy H1 | For absorbance measurement at 450 nm (reference 570 nm); 96-well plate reader |
| Mobile application (dietary record) | Nutritionist Pro | Mobile App v2.1 | For patient self-recording of meals; syncs with nutritionist software |
| Nutritional assessment software | Nutritionist Pro | v7.0 | For calculating BMR, REE, and generating 7-day food exchange plans |
| Personal protective equipment (lab coat, gloves, goggles, mask) | Various | N/A | BSL-2 requirement; latex gloves, nitrile optional; safety goggles with anti-fog |
| Refrigerated centrifuge | Eppendorf | 5810R | Temperature range -9°C to 40°C; fixed-angle rotor F-34-6-38; used for blood sample centrifugation |
| Resistin ELISA kit | R&D Systems | DY1359 | For detection of human resistin (RETN) in serum; intra-assay CV <5% |
| SAA ELISA kit | Abcam | ab100634 | For human serum amyloid A1 (SAA1) measurement; range 0.15–20 ng/mL |
| Saxagliptin tablets (5 mg) | Jiangsu Aosaikang Pharmaceutical | H20193008 | Oral DPP-4 inhibitor; 5 mg once daily; store at 20-25°C |
| Serum separator tubes (yellow/red cap) | BD Vacutainer | 367988 | For serum separation; contains clot activator and gel barrier; 4 mL draw |
| Sharp container | BD | 305790 | Puncture-resistant container for safe disposal of needles and lancets |
| SPSS software | IBM | Version 25.0 | For statistical analysis: t-tests, ANOVA, chi-square; Windows 10 compatible |
| Stadiometer | Seca | 213 | For height measurement; range 60–200 cm; wall-mounted with movable headpiece |
| Standard soft tape measure | Seca | 201 | For waist and hip circumference measurement; accuracy 0.1 cm; non-stretchable |
| Synergy H1 | Microplate reader (ELISA) | BioTek | Synergy H1 |
| TNF-α ELISA kit | R&D Systems | DY210 | For human TNF-α detection; sensitivity <2 pg/mL; 96-well plate |
| Training injection pen (needle-free) | Novo Nordisk | 123456 (sample) | Device for patient training; no needle; replicates handling of real injection pen |
| Vacuum blood collection needle set (21G) | BD Vacutainer | 367281 | Multi-sample needle; 21G × 1.5 inch; for venipuncture with holder |
| Vaspin ELISA kit | R&D Systems | DY1206 | For human Vaspin (SERPINA12) measurement; assay range 0–20 ng/mL |
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