Published online Nov 27, 2026. doi: 10.4240/wjgs.125359
Revised: August 24, 2026
Accepted: October 26, 2026
Published online: November 27, 2026
Processing time: 130 Days and 19 Hours
Intraperitoneal adhesions are fibrotic scars that form between abdominal organs after surgery and a major cause of postsurgical morbidity. Despite their clinical significance, the precise molecular mechanisms underlying their formation remain largely unclear, and standardized therapeutic strategies have yet to be establi
To investigate the role of thrombospondin-1 (Thbs-1) in peritoneal adhesion for
A standardized peritoneal adhesion model was developed. Thbs-1-knockdown mice and HMrSV5 cell lines were established. Twenty-five mice were randomized to five treatment groups: Sham, negative control, sh-Thbs-1, normal saline, and Leu-Ser-Lys-Leu (LSKL). Adhesion severity was assessed 7 days post-surgery by Nair’s score and through immunohistochemistry, multi-immunofluorescence, and oxidative stress assays.
Thbs-1 was found to be highly expressed during adhesion formation. Both sh-Thbs-1 and LSKL treatments sig
Thbs-1 deficiency reduces adhesion formation by inhibiting inflammation and ROS production while promoting mesothelial repair. Accordingly, Thbs-1 represents a promising therapeutic target for postsurgical adhesions.
Core Tip: This study identifies thrombospondin-1 (Thbs-1) as a critical driver of postsurgical intraperitoneal adhesions. We demonstrate that Thbs-1 deficiency, achieved through genetic knockdown or pharmacological inhibition with Leu-Ser-Lys-Leu, significantly attenuates adhesion formation. The underlying mechanism involves the reduction of inflammation and oxidative stress alongside the promotion of peritoneal mesothelial cell repair. By uncovering this Thbs-1-mediated pathway, our findings offer a novel therapeutic strategy to mitigate postsurgical morbidity, highlighting Thbs-1 as a promising target for clinical intervention in abdominal surgery.
- Citation: Tong HC, Ye CC, Zhang ZX, Lin WH, Qu C, Gao YY, Wang JH, Wu YH, Liu YT. Silencing of thrombospondin-1 prevents abdominal adhesion formation by reducing inflammation and oxidative stress. World J Gastrointest Surg 2026; 18(11): 125359
- URL: https://www.wjgnet.com/1948-9366/full/v18/i11/125359.htm
- DOI: https://dx.doi.org/10.4240/wjgs.125359
Intraperitoneal adhesions, a prevalent long-term complication of abdominal surgery, arise in 50%-90% of patients after abdominal or pelvic procedures. A certain proportion of these patients require reoperation, which can result in serious complications such as intestinal obstruction, chronic pain, infertility, and mortality[1,2]. Moreover, adhesions impose a significant socioeconomic burden through prolonged hospitalization and subsequent surgical interventions. Despite various preventive strategies, including anti-inflammatory agents, anticoagulants, fibrinolytic drugs, and barrier materials, no effective clinically approved medications have been developed to date[3]. The limited efficacy of current strategies is primarily due to the challenge of concurrently targeting inflammation, oxidative stress, and impaired fibri
Mesothelial cells (MCs) play a vital function in adhesion prevention by secreting protective molecules[4]. Surgical trauma induces MC damage, reactive oxygen species (ROS) accumulation, and immune cell recruitment[5,6], triggering fibrin deposition and impaired fibrinolysis, which are hallmarks of adhesion pathogenesis[7]. This imbalance is characterized by reduced tissue plasminogen activator activity and elevated plasminogen activator inhibitor-1 expression. Con
Thrombospondin-1 (Thbs-1), a protein involved in tissue repair and remodeling[8], is linked to fibrin clot formation, inflammation, and fibrosis[9]. Thbs-1 modulates inflammatory cytokines[10] and activates transforming growth factor beta (TGF-β), a master regulator of fibrosis[11]. Beyond these roles, Thbs-1 can directly induce ROS production via receptors such as cluster of differentiation 47 (CD47), further amplifying inflammation and peritoneal damage. Given that MCs are the cells most affected by surgical trauma, investigating mesothelial-derived Thbs-1 and its role in oxidative stress and TGF-β activation is crucial for understanding early adhesion initiation. In the present study, Leu-Ser-Lys-Leu (LSKL), a specific inhibitor of Thbs-1-mediated TGF-β activation, was employed to explore these pathways. Although Thbs-1 is implicated in various fibrotic disorders, its specific role in postoperative abdominal adhesions remains poorly understood.
The present study evaluated Thbs-1 expression in adhesion tissues and assessed the impact of Thbs-1 silencing or inhibition on adhesion formation. The results of in vitro and in vivo experiments demonstrate that Thbs-1 is a potential therapeutic target for preventing postoperative abdominal adhesions, offering new perspectives on the pathogenesis and prevention of this condition.
Male C57BL/6 mice (6 weeks old, 20-25 g) were obtained from Beijing Vital River Laboratory Animal Technology Co., Ltd (Beijing, China). Female mice were excluded to minimize variability associated with estrous cycles, pregnancy, and lactation. The animal protocol was designed to minimize pain or discomfort to the animals. The animals were acclimatized to laboratory conditions (23 °C, 12 hours/12 hours light/darkness, 50% humidity, ad libitum access to food and water) for 7 days before the experiment. All procedures were approved by the Ethics Committee of Xi’an Jiaotong University, No. XJTUAE2024-2003 and were conducted in accordance with the Guide for the Care and Use of Laboratory Animals.
Prior to surgery, mice were fasted overnight. Twenty animals were randomly assigned to four treatment groups. Anesthesia was induced with isoflurane. Following disinfection, a 1-1.5-cm midline abdominal incision was made. The ileocecal abrasion procedure involved scraping the anterior surface of the cecum 20-30 times with gauze to induce mild serosal hemorrhage, with a similar abrasion applied to the parietal peritoneum. The incision was closed with 4-0 absorbable sutures. At 1 day, 3 days, 5 days, and 7 days post-surgery, mice were eutha
Lentiviral vectors were obtained from Shanghai Genechem Co., Ltd (Shanghai, China). For in vivo transduction, nine mice were randomly assigned to three groups to receive intraperitoneal injections of either a negative control (NC) lentivirus or one of two Thbs-1-knockdown lentiviral sequences (1 × 109 TU/mL, 50 μL per mouse). The viruses were engineered to carry a firefly luciferase reporter to enable in vivo imaging. Three days post-injection, bioluminescence imaging was performed. Mice received an intraperitoneal injection of D-luciferin sodium (150 mg/kg), and fluorescence signals were acquired 10 minutes later. The Thbs-1-knockdown lentivirus that demonstrated successful transduction was selected for subsequent experiments.
For in vivo experiments, 25 mice were randomly allocated to five groups: Sham operation, NC, Thbs-1 knockdown [short hairpin (sh)-Thbs-1], normal saline (NS), and LSKL. Mice in the sham group underwent laparotomy only, while the remaining four groups were subjected to the same modeling procedure described above. Lentiviral delivery was performed prior to modeling: The sh-Thbs-1 group received Thbs-1-knockdown lentivirus, and the NC group received negative control lentivirus, following the same protocol as described previously. For pharmacological intervention, mice in the LSKL group were administered intraperitoneally at a dose of 30 mg/kg immediately before abdominal wall closure and every other day after surgery. Control mice received an equal volume of NS intraperitoneally at the corresponding time points. This dose and route were adopted based on previously published protocols demonstrating the systemic anti-fibrotic efficacy of LSKL in murine models[12]. No abnormal behavioral changes, weight loss, or signs of toxicity were observed in LSKL-treated mice throughout the experimental period. Seven days after modeling, all mice were euth
For assessment of abdominal adhesions, a 0.2-cm incision was made in the left lower abdominal wall, followed by an inverted U-shaped incision extending from the xiphoid process to the right lower quadrant. The abdominal wall was carefully retracted to avoid damaging any adhesions and thus possibly interfering with adhesion evaluation. Adhesion severity was assessed independently by two researchers using Nair’s adhesion scoring system[13], as follows: (1) Grade 0: No adhesion; (2) Grade I: A single adhesion zone between the viscera or between the viscera and abdominal wall; (3) Grade II: Two adhesion zones between the viscera or between the viscera and abdominal wall; (4) Grade III: More than two adhesion zones, with no direct adhesion of the viscera to the abdominal wall; and (5) Grade IV: Direct adhesion of the viscera to the abdominal wall, regardless of the number of adhesion zones.
Adhesion tissues were fixed in 10% formalin, embedded in paraffin, and sectioned at 4 μm for subsequent analyses. For hematoxylin-eosin (H&E) staining, adhesion tissue sections were dewaxed in xylene and stained with hematoxylin and eosin. Images were acquired using a Leica slide scanning microscope (Leica, Wetzlar, Germany). Inflammatory infiltration was assessed by laboratory professionals using a semiquantitative histopathological scoring system adapted from Hooker et al[14], as follows: 0: No inflammatory cell infiltration; 1: Presence of macrophages, lymphocytes, or plasma cells; 2: Presence of giant cells, plasma cells, eosinophils, and neutrophils; and 3: Inflammatory cell infiltration with microabscess formation. For immunohistochemistry (IHC) analysis, adhesion tissue sections were dewaxed, rehydrated, and subjected to antigen retrieval. Endogenous peroxidase activity was blocked with 3% hydrogen peroxide, followed by blocking with normal serum. The sections were then incubated with primary antibody (anti-Thbs-1, #18304-1-AP, 1:2000, Proteintech, Chicago, IL, United States), visualized using 3,3′-diaminobenzidine (DAB), and counterstained with hematoxylin. Whole-slide images were captured using a microscopic imaging system (3DHISTECH, Shandong Province, China). The percentage of Thbs-1-positive area was quantified using Fiji software by applying a fixed threshold to DAB-stained sections across five random high-power fields per slide.
For Sirius red staining, frozen sections of adhesion tissues were equilibrated to room temperature from -20 °C, fixed with tissue fixative for 15 minutes, and rinsed under running water. Sections were then stained with Sirius red solution (Wuhan Servicebio Biotechnology Co., Ltd., Wuhan, Hubei Province, China) for 8 minutes, dehydrated in absolute ethanol (Guoyao Group Chemical Co., Ltd., China), and cleared in xylene (Guoyao Group Chemical Co., Ltd., China) for 5 minutes.
For collagen thickness measurement, stained sections of adhesion tissue were examined under a light microscope, where collagen fibers appeared red against a yellow background. For each tissue, at least five sections were randomly selected, and five high-power fields were captured per section. The average collagen fiber thickness across all fields was calculated to represent the fiber thickness for each tissue sample.
The malondialdehyde (MDA) assay was performed using a kit from Beijing Solebro Reagent Co. For analysis, fresh tissue specimens were mechanically homogenized on ice. The homogenate was centrifuged at 8000 × g for 10 minutes at 4 °C, and the supernatant was collected and mixed with the MDA working solution. The mixture was incubated at 100 °C in a water bath for 60 minutes, then cooled in an ice-cold water bath, and centrifuged at room temperature for 10 minutes. An aliquot (200 μL) of the resulting supernatant was transferred to a 96-well plate, and absorbance values at 532 nm and 600 nm were measured using a microplate reader. The normalized MDA content was calculated according to the manu
Superoxide dismutase (SOD) activity was determined spectrophotometrically by measuring the inhibition of cytochrome c reduction at 550 nm and 25 °C. Superoxide anions (O2-) were generated using the xanthine/xanthine oxidase system. SOD competes for superoxide, thereby decreasing the rate of cytochrome c reduction. One unit of SOD activity was defined as the amount of enzyme required to inhibit the rate of cytochrome c reduction by 50%.
Catalase (CAT) activity was analyzed by monitoring the decrease in absorbance at 240 nm and 25 °C in a reaction mixture containing 10 mmol/L H2O2 in phosphate buffer (pH = 7.0). The rate of absorbance decline is proportional to CAT activity.
Glutathione peroxidase (GPx) activity was measured according to a coupled reaction with glutathione reductase. The assay quantifies the rate of oxidation of reduced glutathione by monitoring the decrease in NADPH concentration upon addition of glutathione reductase to the reaction mixture.
The human peritoneal mesothelial cell line HMrSV5 was obtained from the Shanghai Institute of Cell Biology, Chinese Academy of Sciences. Cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM, Gibco, Thermo Fisher Scientific, Waltham, MA, United States) supplemented with 10% (v/v) fetal bovine serum (FBS, HyClone, GE Healthcare Life Science, Pittsburgh, PA, United States) and 100 U/mL penicillin-streptomycin, at 37 °C in a 5% CO2 humidified incubator. To establish stable Thbs-1-knockdown HMrSV5 cell lines, lentiviral vectors (Shanghai GenePharma Co., Ltd., Shanghai, China) were transduced into HMrSV5 cells. After 72 hours, stably transduced cells were selected with 5 μg/mL puromycin (Sigma-Aldrich, Shanghai, China).
Supernatants from HMrSV5 cells were collected and centrifuged at 4 °C and 3000 rpm for 15 minutes. The clarified supernatant was then carefully transferred, and the concentration of Thbs-1 protein was analyzed using an enzyme-linked immunosorbent assay (ELISA) kit (MultiSciences, Hangzhou, Zhejiang Province, China) according to the manufacturer’s instructions. Each treatment was performed in triplicate. Absorbance at 450 nm was measured using a microplate reader within 10 minutes after reaction completion, after zeroing with a blank control, and Thbs-1 protein concentrations were quantified accordingly.
Total RNA was extracted from cells using TRIzol reagent (Invitrogen, CA, United States). Complementary DNA (cDNA) was synthesized using the Thermo Scientific RevertAid First Strand cDNA Synthesis Kit (Thermo Fisher Scientific, CA, United States). Quantitative real-time polymerase chain reaction (PCR) was performed using iQ™ Supermix (Bio-Rad, CA, United States). Relative mRNA expression levels were calculated using the 2-ΔΔCt method. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene expression was measured as the control. The primer sequences used in this study were: Thbs-1 (F: 5’-AGACTCCGCATCGCAAAGG-3’ and R: 5’-TCACCACGTTGTTGTCAAGGG-3’), GAPDH (F: 5’-TGCACCACCAACTGCTTAGC-3’ and R: 5’-GGCATGGACTGTGGTCATGAG-3’).
Cell viability was assessed using the Cell Counting Kit-8 (CCK-8, 7 Sea Biotech, Shanghai, China). HMrSV5 cells were seeded in 96-well plates at 5000 cells per well and cultured for 3 days with or without TGF-β. Then the CCK-8 assay was performed according to the manufacturer’s protocol.
HMrSV5 cells were seeded in 6-well plates and cultured to confluence. A scratch wound was created across the cell monolayer using a 200-μL pipette tip. After 48 hours in culture at 37 °C with 5% CO2, images were captured using a photomicroscope (Leica, Wetzlar, Germany). The cell migration ratio was quantified using Scion Image software.
For in vivo analysis of ROS production, frozen adhesion tissues and normal peritoneum were sectioned at 4 μm, air-dried at room temperature, and encircled with a hydrophobic barrier pen. After washing, an autofluorescence quencher was applied. ROS staining solution (Sigma-Aldrich, Shanghai, China) was added to the encircled areas and incubated at 37 °C for 30 minutes in darkness. Nuclei were counterstained with DAPI (Wuhan Servicebio Technology Co., Ltd., Wuhan, Hubei Province, China) for 10 minutes at room temperature in the dark. Sections were then mounted with an anti-fluorescence quenching mounting medium and imaged using a fluorescence microscope (Olympus, Japan). For each tissue sample, five high-power fields were randomly selected per section, and fluorescence intensity in each field was analyzed using Fiji software. The average fluorescence intensity across all fields was calculated to represent the ROS level for each tissue sample.
For in vitro analysis of ROS production, HMrSV5 cells were seeded in 6-well plates and treated with or without H2O2 (500 μmol/L). Following treatment, the HMrSV5 cells were incubated with the ROS dye (Beyotime Biotechnology, China) for 20 minutes at 37 °C. ROS levels were then assessed by fluorescence microscopy and quantified by flow cytometry (BD FACS Celesta™). The data were analyzed using Fiji and FlowJo software.
Statistical analyses were performed using SPSS 18.0 (SPSS Inc., Chicago, IL, United States) and GraphPad Prism software (GraphPad, San Diego, CA, United States). Data are presented as n (%), or mean ± SD. Normality was assessed using the Shapiro-Wilk test. Normally distributed data were analyzed using Student’s t-test (two-tailed) or one-way analysis of variance, whereas non-normally distributed data were analyzed using the Kruskal-Wallis test, with Dunn’s post-hoc test applied for multiple comparisons. Categorical and ordinal data, including Nair’s adhesion scores and inflammatory scores, were appropriately analyzed using non-parametric tests given their ranked, non-continuous nature. Qualitative data were analyzed using the χ² test or Fisher’s exact test. Two-tailed P values < 0.05 were considered statistically sig
Mice were euthanized at 1 day, 3 days, 5 days, and 7 days after peritoneal injury and examined for adhesion formation (Figure 1). The results showed that adhesions were visible 3 days after the induction of peritoneal injury and appeared as soft attachments between organ surfaces (Figure 1A). Dense adhesion bands formed between the cecum and the abdominal wall over time, resulting in significant increases in Nair’s scores (Figure 1D). Inflammation is the initiating factor in the formation of abdominal adhesion; thus, we assessed the inflammatory status of the adhesion tissue. H&E staining of the adherent tissues confirmed the infiltration of inflammatory cells in the adherent tissues and the increase in the inflammatory scores over time (Figure 1B and E).
As a synergistic molecule of TGF-β1, Thbs-1 is involved in a number of biological processes such as inflammation, stress, and fibrosis. A previous global microarray analysis showed that Thbs-1 is highly expressed in MCs after artificial induction of mesothelial-mesenchymal transition using TGF-β1[15]. However, the role of Thbs-1 in postoperative abdominal adhesion formation and its mechanisms have not been elucidated[5]. In the present study, we examined the expression of Thbs-1 in postoperative abdominal adhesion tissues by IHC and found that Thbs-1 expression was significantly higher on days 3-7 compared with day 1 postoperatively (Figure 1C and F).
To validate whether Thbs-1 expression is a reason for mesothelial cell injury in the process of adhesion formation, we created adhesions in wild-type mice that had been locally treated with lentivirus-mediated Thbs-1 knockdown during adhesion formation. Next, in vivo small animal imaging was performed to select stable Thbs-1-knockdown mice. The fluorescence intensities in the sh-Thbs-1#2 transfected mice were significantly higher than those in the NC group, and thus, these mice were selected for subsequent use (Figure 2A). Then we examined Thbs-1 expression in the adhesion tissues of mice on day 7 after induction of peritoneal injury. Consistent with the former results, IHC staining and quantification confirmed that the relative expression of Thbs-1 was down-regulated in the sh-Thbs-1 group (Figure 2B). We then assessed the effect of LSKL, a functional inhibitor of Thbs-1. IHC analysis showed a trend of decreased Thbs-1 protein expression in the LSKL-treated group, although the change was not statistically significant (Figure 2B). This result is consistent with the known mechanism of LSKL as a functional inhibitor that primarily blocks the biological activity of the Thbs-1 protein rather than suppressing its expression. Crucially, and consistent with the effects of Thbs-1 knockdown, LSKL treatment significantly attenuated postoperative abdominal adhesion formation, as reflected by the markedly lower Nair’s scores in comparison with those of the NS group (Figure 2C). This result demonstrates that functional inhibition of Thbs-1 is sufficient to mitigate adhesion.
Inflammation and stress are key steps in the formation of abdominal adhesions, and therefore, we further explored the effects of Thbs-1 inhibition on inflammation and oxidative stress (Figure 3). H&E staining of the adherent tissues showed that adhesive bands in mice with Thbs-1 knockdown or in mice treated with LSKL were sparse relative to those in controls, and inflammatory scores in these mice were reduced (Figure 3A and C). Next, we conducted multi-immunofluorescence staining to validate the presence of oxidative stress in the adhesion tissues (Figure 3B). The results showed that the intracellular red fluorescence intensity was significantly reduced in the sh-Thbs-1 group or after exposure to LSKL, suggesting a decrease in ROS content. Afterwards, we examined changes in key enzymes during oxidative stress in adhesion tissues. These results showed that the MDA levels in the sh-Thbs-1 and LSKL-treated groups were significantly lower than those in the NC and NS groups, and the expression levels of SOD and GPx were significantly higher than those in the NC and NS groups (Figure 3D-F).
Peritoneal surface repair differs from traditional wound healing. A restored peritoneal surface after surgical trauma involves interactions of surrounding cells, with regeneration of the mesothelial surface and without scar formation. The extent of peritoneal mesothelial cell damage was demonstrated using the peritoneal mesothelial cell marker CK19 labeling, which showed that the sh-Thbs-1 and LSKL-treated groups had lower rates of peritoneal mesothelial cell integrity than the control groups (Figure 4A). Collagen fiber formation disrupts peritoneal mesothelial cell repair, and thus, we used Sirius red staining to assess collagen fiber formation in adhesion tissue. The results confirmed that the collagen fiber thicknesses were lower in the sh-Thbs-1 and LSKL-treated groups than in the NC and NS groups (Figure 4B and C).
Thbs-1 is a molecule secreted extracellularly to mediate cell adhesion and as such plays an important role in the healing of injured tissue. To determine whether Thbs-1 deficiency is a protective factor in HMrSV5 cells, we constructed a HMrSV5 cell line with stable knockdown of Thbs-1. ELISA experiments showed that the concentration of Thbs-1 protein in the supernatant was significantly reduced after Thbs-1 knockdown (Figure 5A), and the relative expression of Thbs-1 mRNA in the cell pellet was also significantly reduced (Figure 5B). The rapid proliferation and migratory adhesion of peritoneal MCs are critical factors in the formation of the initial ‘fibrin bridge’ and the progression of abdominal adhesions. Specifically, TGF-β1-induced rapid proliferation of peritoneal MCs was significantly accelerated after induction with TGF-β1, and this effect was reversed by knockdown of Thbs-1 (Figure 5C). To determine whether sh-Thbs-1 modulates the migratory adhesion between damaged surfaces, a cell scratch assay was performed. The results showed that HMrSV5 cells on both sides of the wound showed significant adhesion after 24 hours, whereas sh-Thbs-1 treatment significantly slowed down this migratory adhesion across the wound (Figure 5D and E). Subsequently, to determine whether Thbs-1 knockdown protects MCs from H2O2-induced oxidative stress, we measured the ROS level in NC and sh-Thbs-1 cells. In this assay, the intracellular green fluorescence intensity was significantly enhanced under H2O2 stimulation, whereas in the sh-Thbs-1 group, this increase in ROS was significantly attenuated (Figure 5F and G).
Current clinical approaches for alleviating abdominal adhesion include physical, medical, and surgical interventions. However, these strategies are often associated with limited efficacy, prolonged treatment duration, recurrence, and risks of serious complications[16]. Thus, there is a pressing need to develop more effective methods for the prevention and treatment of abdominal adhesions. In this study, we investigated the potential involvement of Thbs-1 in the pathogenesis of abdominal tissue adhesion and conducted experiments to elucidate the underlying mechanisms. Thbs-1 is an adhesive glycoprotein originally identified in platelets. Previous studies have shown that Thbs-1 promotes the migration and proliferation of vascular endothelial cells and hypertrophic scar fibroblasts, and shows significantly upregulated exp
Inflammation serves as the initiating event in abdominal adhesion formation[23,24]. Thbs-1, which is secreted in response to inflammatory stimuli, has been implicated in tissue injury, inflammatory diseases, and fibrosis[25,26]. Ana
Under normal conditions, the mesothelium reestablishes an intact and functional nonadhesive layer within a few days after injury. However, peritoneal MCs are lost following injury, and the cells responsible for tissue repair are derived from adjacent normal peritoneal cells or other stem cells. In an inflammatory environment, these reparative cells may undergo necrosis or differentiate into fibroblasts[34,35]. Therefore, accelerating the repair process may help prevent adhesion formation. In the present study, mice in the sh-Thbs-1 group exhibited reduced collagen fiber thickness on the peritoneal surface after surgical trauma, as well as a lower rate of peritoneal mesothelial cell integrity. In vitro, the wound healing area (migration rate) of sh-Thbs-1 MCs was significantly decreased compared with that of control cells. Together, these findings suggest that Thbs-1 deficiency promotes peritoneal mesothelial cell repair and strongly support that tar
Thbs-1 exerts complex, context-dependent effects on inflammation and tissue repair by modulating cell adhesion, activating latent TGF-β, and interacting with CD47. Our findings are consistent with those of Liu et al[35], who reported that Thbs-1 deficiency suppresses vascular inflammation by inhibiting monocyte motility. However, Tai et al[36] demon
Several limitations of the present study should be noted. First, the translational relevance of these mouse model findings to human patients requires clinical validation. Future work will prospectively collect archived human adhesion specimens for Thbs-1 immunohistochemistry and pathway validation. Second, intraperitoneal lentivirus delivery lacks cell-type specificity, and the contributions of other Thbs-1-expressing cell types, such as macrophages, fibroblasts, and endothelial cells, could not be distinguished. Future studies using cell-specific conditional models are warranted. Finally, whether the observed short-term efficacy of Thbs-1 inhibition is sustained over extended periods remains to be rigorously validated. Additionally, while the inhibitory effect of LSKL supports the possible involvement of the Thbs-1-TGF-β axis in abdominal adhesion formation, the present study did not directly measure active TGF-β levels in adhesion tissues or assess the phosphorylation status of downstream Smad2/3 across the different experimental groups. The proposed role of Thbs-1-mediated TGF-β signaling should consequently be interpreted with caution. Future studies incorporating direct quantitative measurements of active TGF-β and phosphorylated Smad2/3, together with pathway-specific rescue or inhi
In this study, we observed marked upregulation of Thbs-1 in adherent abdominal tissues. Thbs-1 deficiency effectively mitigated adhesion formation in vivo by attenuating inflammation and ROS accumulation while also promoting peri
This study was completed in the Shaanxi Provincial Key Laboratory of Infection and Immune Diseases; the authors would like to thank all laboratory staff for their help with the experiments.
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