Open Access

 Protective effect of scutellarin on myocardial cells treated with high glucose

  • Authors:
    • Xiaojuan Wei
    • Zhigang Li
    • Shude Li
    • Jun Yang
    • Biao Fan
    • Shuiwang He
    • Siman Li
  • View Affiliations

  • Published online on: June 19, 2025     https://doi.org/10.3892/br.2025.2021
  • Article Number: 143
  • Copyright: © Wei et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Diabetic cardiomyopathy (DCM) is an important cause of death in patients with diabetes. DCM can be simulated by cardiomyocyte injury induced by high glucose (HG) in vitro. Scutellarin (Scu) is a flavonoid extracted from Erigeron breviscapus. The H9c2 cell line was used as an in vitro model in the present study to investigate the mechanism by which Scu reduces HG‑induced cardiomyocyte injury. Moreover, the present study aimed to provide scientific evidence on the mechanism by which Scu prevents DCM. The following groups were used: Control, model, Scu (50/100/200/400 µM) and curcumin. Following H9c2 cell adherence, the control and model groups were treated with normal medium; the Scu group cells were treated with different concentrations of Scu, whereas the curcumin group cells were treated with 4 µM curcumin for 4 h. Subsequently, the normal group was cultured in normal medium, and the other groups were treated with medium containing 100 mM HG for 48 h. The results indicated that Scu improved the morphology of H9c2 cells treated by HG, enhanced cell proliferative activity, reduced the production of reactive oxygen species and the induction of apoptosis. Moreover, Scu promoted the expression of Bcl‑2 and inhibited the expression levels of caspase‑3, cleaved caspase‑3, caspase‑9, cleaved caspase‑9, caspase‑12, Bax, NADPH oxidase (Nox)2 and Nox4. The findings indicated that Scu could inhibit oxidative stress and reduce the induction of apoptosis in cardiomyocytes, thereby alleviating HG‑induced myocardial injury.

Introduction

Economic development promotes the continuous improvement of the standard of living. Diabetes mellitus (DM) is one of the major chronic diseases that endanger human health. Type 2 diabetes mellitus (T2DM), which is mainly caused by insulin resistance (IR), accounts for >90% of DM. Long-term IR often causes damage to heart function, a condition known as diabetic cardiomyopathy (DCM). DCM causes extensive focal myocardial necrosis, subclinical cardiac dysfunction, and eventually progresses to heart failure, arrhythmias, cardiogenic shock and even sudden death in patients with severe disease. DCM has become one of the leading causes of death in diabetic patients. The main features of DCM are oxidative stress and myocardial hypertrophy, apoptosis, inflammation and fibrosis (1). Previous research has shown that ~12% of diabetic patients suffer from DCM, which further develops into heart failure and eventually leads to death (2). Apoptosis is one of the main causes of myocardial injury. In the myocardial injury caused by T2DM, the activity of caspase-3 is increased in myocardial cells, activating the apoptotic pathway by death receptors and leading to myocardial cell apoptosis (3,4). Under high glucose (HG) conditions, the content of reactive oxygen species (ROS) in myocardial cells increases, leading to an increase in caspase-12 levels, which in turn activates the apoptotic pathway of endoplasmic reticulum stress (5). Therefore, the process of cell apoptosis in myocardial injury caused by long-term diabetes is the main cause of heart failure.

Scutellarin (Scu) is the most abundant flavonoid extracted from Erigeron breviscapus (6,7). The IUPAC name is 4,5,6-trihydroxyflavone-7-glucosidic acid. The appearance of the compound is a yellow powder, which is difficult to dissolve in water. Previous studies have shown that Scu exhibits optimal therapeutic effects on a variety of cancer cell types, without apparent toxicity or side effects (8-11). Long et al (12) demonstrated that Scu could inhibit angiogenesis in diabetic retinopathy by downregulating the VEGF/ERK/FAK/Src signaling pathway, and Wang et al (13) revealed that Scu can alleviate liver injury in T2DM rats by downregulating the levels of homocysteine (13). Scu has also been shown to reduce the accumulation of free radicals by scavenging oxygen free radicals, thereby reducing the damage of free radicals to the body (14). Scu also exhibits a significant protective effect on myocardial ischemia in type II diabetic rats and can inhibit the overexpression of cardiomyocyte-related proteins caused in an HG environment (15,16). In vivo experiments have also shown that Scu can protect against heart injury in diabetic patients by reducing oxidative stress, inflammatory reactions and apoptosis (17,18). Therefore, the present study aimed to explore whether inhibition of oxidative stress-related factors can reduce apoptosis in vitro, thereby providing a potiential target and theoretical foundation for the clinical treatment of DCM.

Several studies have confirmed the establishment of an HG-induced cardiac injury model using the H9c2 cell line (19-21). Therefore, in the present study, the H9c2 cardiomyocyte cell line was used to establish an in vitro model. The expression levels of major factors related to cardiomyocyte apoptosis and oxidative stress were investigated by detecting relevant indicators, to determine whether Scu can reduce cardiomyocyte apoptosis caused by HG by inhibiting oxidative stress-related factors. This may provide a new theoretical basis and potential target for the use of Scu in reducing HG-induced myocardial injury in diabetes.

Materials and methods

Cell culture and treatment protocols

The H9c2 cell line was purchased from the Cell Bank of the Chinese Academy of Sciences and cultured in DMEM with low NaHCO3 (iCell Bioscience Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) and 100 U/ml penicillin and streptomycin mixture (Gibco; Thermo Fisher Scientific, Inc.) in an atmosphere of 5% CO2 at 37˚C. The medium was replaced every 2 days, and the cells were digested with 0.25% trypsin (Gibco; Thermo Fisher Scientific, Inc.) when their density reached 80-90%. H9c2 cells were seeded in 6-well plates or 96-well plates for the following experiments.

The experiment was divided into seven groups, including the following: Control (no treatment), model (HG treatment for 48 h), Scu 50 µM treatment, Scu 100 µM treatment, Scu 200 µM treatment, Scu 400 µM treatment, and curcumin treatment (4 µM; positive control group). The density of the H9c2 cell line reached 90% following 24 h of culture in an atmosphere of 5% CO2 at 37˚C. The cell culture medium of the control group and the model groups was replaced with normal medium; the treatment groups with different concentrations of Scu were replaced with medium containing Scu (50/100/200/400 µM), and the curcumin group was replaced with medium containing 4 µM curcumin; the incubation was performed for 4 h in an atmosphere of 5% CO2 at 37˚C. Following the 4 h, the control group continued to be cultured with normal medium and the other treatment groups were replaced with medium containing 100 mM HG for 48 h.

The experiment investigating the effects of APX-115 on H9c2 cardiomyocytes was divided into four groups, including the following: The control group (no treatment), the APX-115 group, the HG group, and the APX-115 + HG group. The density of the H9c2 cell line reached 90% following 24 h of culture in an atmosphere of 5% CO2 at 37˚C. The cell culture medium of the control group and the HG group was replaced with normal medium, and the APX-115 group and the APX-115 + HG group were replaced with medium containing 5 µM APX-115 free base; the incubation was performed for 2 h in an atmosphere of 5% CO2 at 37˚C. After the 2 h, the cell culture medium of the control group and the APX-115 group was replaced with normal medium, and the HG group and the APX-115 + HG group were replaced with medium containing 100 mM HG for 48 h.

Reagents and antibodies

Scu (purity: 98%) was donated by the Pharmacy of Kunming Medical University. It was dissolved in PBS and prepared into mother liquor for subsequent use. The antibodies used for detecting GAPDH (cat. no. AF7021), caspase-3 (cat. no. AF6311), cleaved caspase-3 (cat. no. AF7022), cleaved caspase-9 (cat. no. AF5240), Bax (cat. no. AF0120) and Bcl-2 (cat. no. AF6139) were obtained from Affinity Biosciences, those used for caspase-9 (cat. no. bs-0049R), NADPH oxidase (Nox)2 (cat. no. bs-3889R) and Nox4 (cat. no. bs-1091R) were acquired from BIOSS and the antibodies for caspase-12 (cat. no. Abs120719) were obtained from Absin (Shanghai) Biotechnology Co., Ltd. The Cell Counting Kit-8 (CCK-8) assay kit was obtained from Biosharp Life Sciences, the Annexin V-633 Apoptosis assay kit (cat. no. AD11) was acquired from Dojindo Laboratories, Inc. and the Total Reactive Oxygen Species (ROS) Assay Kit 520 nm (cat. no. 88-5930) was purchased from Thermo Fisher Scientific, Inc. The Total Reactive Oxygen Species (ROS) Assay Kit 520 nm contained the necessary reagent and buffer for identifying ROS in cells by flow cytometry in the FITC channel. Nox inhibitor APX-115 free base (5 µM; cat. no. HY-120801A) was obtained from MedChemExpress.

CCK-8 assay for the detection of the cell proliferative activity

The original medium of the 96-well plate was discarded and 100 µl of pre-prepared medium containing CCK-8 (CCK8: medium=1:10) was added to each well. Following incubation at 37˚C for 4 h, the optical density (OD) value was measured at 450 nm. The proliferative activity of the cells in each group was calculated according to the following formula of cell proliferation activity:

Detection of the levels of ROS using flow cytometry

H9c2 cells were incubated into 6-well plates at a cell density of 5x105 cells/ml. Following 24-h incubation, the medium was discarded, washed 3 times with PBS, and digested with trypsin. The cells were collected and centrifuged at 93 x g for 5 min at room temperature (22-25˚C), and the supernatant was discarded. Subsequently, the cells were resuspended and washed twice with PBS. The supernatant was discarded and 100 µl of 1X ROS staining (1,000 µl ROS assay buffer +2 µl 500X ROS assay stain) was added to each group, resuspended and then cultured in a CO2 incubator at 37˚C for 1 h. The cells were centrifuged at 93 x g for 5 min at room temperature (22-25˚C), the supernatant was discarded and the cells were washed with PBS. Following centrifugation again, the supernatant was discarded and resuspended in 1 ml of PBS. The levels of ROS were detected by flow cytometry (BD FACSCelesta 3 flow cytometer; BD Biosciences). The average fluorescence intensity was obtained by analysis with Flowjo software (Flowjo 7.6; BD Biosciences), and depicted as a bar chart to compare each group.

Detection of cell apoptosis using flow cytometry

H9c2 cells were incubated into 6-well plates at a density of 5x105 cells/ml. Following 24 h of incubation, the old medium was removed. The cells were washed three times with PBS and the PBS was then removed. The cells were digested with trypsin and collected. Following centrifugation at 93 x g for 5 min at room temperature (22-25˚C), the supernatant was discarded, the cells were resuspended in PBS and washed twice. The cells were resuspended in 100 µl of 1X Annexin V binding solution and 5 µl of Annexin V-633 allowed to mix and 5 µl of propidium iodide solution was added. The culture was kept in the dark for 15 min at room temperature. Subsequently, 900 µl of 1X Annexin V Binding Solution was added to each tube, resuspended and assessed within 1 h. Cell apoptosis was then detected using BD FACSCelesta 3 flow cytometer (BD Biosciences) and the results were analyzed using Flowjo software (Flowjo 7.6; BD Biosciences). The proportion of apoptotic and necrotic cells in the Q2 + Q3 region of the quadrant chart was calculated.

Western blotting

Total protein was extracted with RIPA lysis buffer (cat. no. 89900, Thermo Fisher Scientific, Inc.) supplemented with PMSF (cat. no. 36978; Thermo Fisher Scientific, Inc.), quantified by the bicinchoninic acid method and concentrated to a final density of 3 µg/µl. The loading amount of each protein group was 30 µg and the protein was separated by electrophoresis using a SDS (10%) gel. When the marker was apparently separated, according to the positions of the protein bands with different molecular weights of marker, the gel was cut horizontally and the target strip was in the middle of the gel. Subsequently, the gel was placed in 1X WB transfer buffer (cat. no. D1060-500 ml; Beijing Solarbio Science & Technology Co., Ltd.). Polyvinylidene difluoride (PVDF) membranes were incubated in methanol (30 sec to 60 sec) and subsequently in 1X WB transfer buffer. The required three layers of the filter paper were placed and the sponge pad was incubated into the 1X membrane transfer solution. The sponge pad was placed to three layers of filter paper, a PVDF film was placed on top of the filter paper and the gel with the target strip and three additional layers of filter paper were placed to complete the protein transfer; a semi-dry membrane transfer device was used. The electrode plate and the safety electrode were covered and the electrode was connected to the power supply. Electrophoresis was run at a constant voltage (22V) for protein band transfer. The proteins were transferred to the PVDF membranes and blocked with 5% milk at 37˚C for 40 min. Subsequently, caspase-3, cleaved caspase-3, caspase-9, cleaved caspase-9, caspase-12, Bax, Bcl-2, Nox2 and Nox4 antibodies were incubated overnight at 4˚C at a 1:1,000 dilution. The secondary antibody used was an anti-rabbit (cat. no. ZB-2301; ZSGB-BIO) or anti-mouse (cat. no. ZB-2305; ZSGB-BIO) IgG (diluted 1:10,000) conjugated with horseradish peroxidase. Secondary antibody incubation was performed slowly at room temperature (22-25˚C) on a shaker for 2 h. The ChemiDocTM MP all-round imaging System (Bio-Rad Laboratories, Inc) was used for visual detection. Image Lab 6.0 software (Bio-Rad Laboratories, Inc.) was used to quantify signal intensity.

Immunofluorescence assay

H9c2 cells were incubated into 24-well plates at a cell density of 5x104 cells/ml. Following 24-h incubation, the medium was removed and the cells were washed three times with PBS. Subsequently, 4% paraformaldehyde (fixing solution) was added to the cells and fixed for 20 min at room temperature (22-25˚C). The fixing solution was then removed and the cells were washed with PBS. Subsequently, 0.3% Triton X-100 (permeable liquid) was added and incubated for 20 min at room temperature. The permeable liquid was removed and the cells were washed with PBS. Subsequently, the cells were blocked with FITC-conjugated 5% bovine serum albumin (cat. no. SF163-300 µl; Beijing Solarbio Science & Technology Co., Ltd.) for 2 h at room temperature. The blocking solution was removed and the cells were washed with PBS. The primary antibodies (GAPDH, caspase-3, cleaved caspase-3, caspase-9, cleaved caspase-9, caspase-12, Bcl-2, Bax, Nox2 and Nox4; diluted according to the manufacturer's instructions, 1:200) were incubated at 4˚C overnight (generally 12-16 h). The following day, the cells were washed three times with PBS (all subsequent steps were performed in the dark). A fluorescent secondary antibody [goat anti-rabbit IgG (H+L) Fluor488-conjugated (cat. no. S0018; Affinity Biosciences), diluted according to the instructions provided by the manufacturer, 1:200) was added and incubated for 1 h at room temperature (22-25˚C). Following washing of the cells with PBS, 4',6-diamidino-2-phenylindole was added dropwise and incubated for 5 min in the dark; the cells were washed with PBS one more time. Finally, all cell slices were sealed and the images were observed using a fluorescence microscope. Image Lab 6.0 (Bio-Rad Laboratories, Inc.) software was used to quantify the signal intensity.

Statistical analysis

The data were analyzed using SPSS V17.0 (IBM Corp.) or GraphPad Prism 6.0 software (Dotmatics) and expressed as the mean ± standard error of the mean (SEM). The independent sample t-test was used for comparisons between two groups of data and one-way analysis of variance (ANOVA) was used for comparisons between more than two groups. Tukey's post hoc test was performed to determine the specific group differences following ANOVA. P<0.05 and P<0.01 were considered to indicate a statistically signigicant difference. GraphPad Prism 6.0 software was used for drawing the figures.

Results

An HG-induced cardiac injury model is established using the H9c2 cell line

By observing the cell morphology, the detection of the cell proliferative activity, ROS production and apoptosis, and the detection of the expression level of related proteins, a model of H9c2 cells induced by HG was established. The results indicated that compared with the control group, the number and the density of cells in the model group was decreased and the cell morphology was also altered with an irregular shape (Fig. 1A). The results of the CCK-8 and flow cytometric assays indicated that compared with the control group, the proliferative activity of the cells in the model group was significantly reduced and ROS production and the apoptosis rate were significantly increased (Fig. 1B-D). In the model group, the protein expression levels of the main apoptosis-related factors (caspase-3, caspase-9, caspase-12), certain active fragments (cleaved caspase-3 and cleaved caspase-9) and the key genes for ROS synthesis (Nox2 and Nox4) were upregulated (Figs. 2, 3, 4, 5, 6 and 7). These results indicated that an HG-induced cardiac injury model was established using the H9c2 cell line.

Scu alleviates H9c2 cell injury induced by HG

Firstly, the ability of Scu to reduce the damage of HG to H9c2 cells was determined by observing cell morphology and detecting cell proliferative activity. The results indicated that compared with the model group, the shape and number of the H9c2 cells in the treatment groups of various concentrations of Scu gradually recovered, and the cells were gradually distributed evenly and arranged closely (Fig. 1A). In addition, the cell proliferative activity of each concentration of the Scu treatment group was gradually increased in a dose-dependent manner (Fig. 1B). These results indicated that Scu could alleviate the damage to H9c2 cells induced by HG.

Scu inhibits the expression of the family of apoptotic enzymes (caspases)

Subsequently, the induction of apoptosis of H9c2 cells and the protein expression levels of the apoptosis-related factors was analyzed. Firstly, apoptosis was detected by flow cytometry. The results indicated that compared with that of the model group, the apoptotic rate of each Scu treatment group was significantly decreased (P<0.01) in a dose-dependent manner (Fig. 1C). In addition, the protein expression levels of the apoptosis-related factors were detected using western blotting and immunofluorescence. The results indicated that Scu inhibited the protein expression levels of caspase-3, cleaved caspase-3, caspase-9, cleaved caspase-9 and caspase-12 (Figs. 2, 3, 4 and 5). Moreover, Bax and Bcl-2 are also closely related to apoptosis. The results indicated that Scu inhibited the protein expression levels of Bax and promoted the protein expression of Bcl-2 in a dose-dependent manner (Figs. 2 and 6). These results indicated that Scu inhibited the induction of apoptosis of H9c2 cells induced by HG.

Scu inhibits oxidative stress

Since apoptosis is closely related to the excessive production of ROS, the effect of ROS was further studied on H9c2 cells treated by HG in the presence of Scu. The results of the flow cytometric analysis indicated that compared with the model group, the ROS content of each concentration treatment group of Scu was significantly decreased (P<0.01; Fig. 1D). In addition, the protein expression levels of Nox2 and Nox4, the key factors for ROS production, were assessed using western blotting and immunofluorescence analysis. The results indicated that Scu could inhibit the protein expression levels of Nox2 and Nox4 (Figs. 2 and 7). These results indicated that Scu could reduce H9c2 cell injury induced by HG by inhibiting oxidative stress.

APX-115 free base inhibits Nox overexpression

Finally, the Nox inhibitor APX-115 free base was used to investigate the changes in the related detection indices of the H9c2 cells following inhibition of Nox overexpression. The results indicated that compared with HG-treated cells, the addition of APX-115 increased the number and the density of H9c2 cells, and the cells returned to spindle shape (Fig. 8A), whereas it exhibited a minimal effect on cell proliferative activity and ROS levels (Fig. 8B and C). In addition, western blotting and immunofluorescence results indicated that the protein expression levels of Nox2 and Nox4 in the cells treated with APX-115 free base and HG were downregulated compared with those of the cells treated with HG alone (Fig. 9). These results suggest that Nox inhibitors can inhibit Nox overexpression, thereby alleviating cardiomyocyte injury induced by HG.

Discussion

Apoptosis and oxidative stress are the main features of cardiomyocyte injury in DCM. Caspase is a protease that promotes apoptosis and plays an important role in this cellular process. Various substrates for caspase-3 have been identified, most of which can be hydrolyzed by caspase-3, indicating that the latter plays an important role in the process of apoptosis (22,23). Under normal conditions, caspase-9 exists in a monomeric form in the body; however, when the body is stimulated, caspase-9 can be activated and become a dimer (24). Caspase-12 is mainly concentrated on the surface of the endoplasmic reticulum (ER) and regulates apoptosis via the ER stress response. Therefore, caspase-12 activation is a landmark factor in the apoptotic pathway of ER stress. Previous studies have reported that in C2C12 cells, caspase-12 is activated in response to ER stress; therefore the activation of caspase-9 occurs, which in turn activates caspase-3, caspase-6 and caspase-7 and finally induces apoptosis (25). Similar to caspase-12, the Bcl-2 family of proteins can also mediate apoptosis caused by ER stress involved in cardiovascular diseases (26). Manaenko et al (27) demonstrated that a decrease in the expression levels of the proapoptotic protein Bax could lead to the upregulation of Bcl-2 expression, thereby improving apoptosis in brain injury.

ROS are common oxygen metabolites present in the body and play an important role. During normal oxygen metabolism, ROS can maintain cell homeostasis by coordinating host defense, cell growth and signaling. However, when the body suffers from environmental stress, the production of ROS will increase rapidly, damaging mitochondria and DNA in the cells and thus causing irreversible damage to the normal cells of the body, a phenomenon called oxidative stress (28,29). Nox is one of the major enzymes that produces ROS, and it is an important inflammatory mediator against invading bacteria. Excessive Nox-mediated ROS production in the body leads to a variety of chronic diseases (30). Nox2 and Nox4, as the main enzymes inducing ROS production in cardiomyocytes, play an important role in Ca2+ regulation, mitochondrial regeneration, generation of massive levels of ROS and the inflammatory response following myocardial injury (31,32).

APX-115 free base (3-phenyl-1-(pyridine-2-yl)-4-propyl-1-5-hydroxypyrazole HCl) is an orally active, potent, nonselective Nox inhibitor that effectively inhibits Nox1, Nox2 and Nox4, presumably by broadly mediating the inhibition of various Nox isoforms (33). Previous studies have shown that the APX-115 free base can improve kidney and HG-induced inflammation and fibrosis in mouse foot cell lines, improve IR in diabetic mice, and reduce urinary protein and plasma creatinine levels by inhibiting Nox gene upregulation and protein expression levels (33,34). The experiments of Kwon et al (35) have also confirmed that APX-115 free base can effectively prevent renal injury, such as proteinuria, glomerular hypertrophy, renal tubular injury, podocyte injury, fibrosis, inflammation and oxidative stress, in diabetic mice.

Individuals with diabetes often do not produce enough insulin, cannot produce enough insulin, or cannot use insulin effectively, causing blood glucose levels to rise (36). The prolonged HG environment in the body can cause vascular damage (37), which in turn leads to damage to heart function (38). This damage often manifests as oxidative stress in heart muscle cells (39), myocardial hypertrophy, cell apoptosis (40), inflammation (41), and myocardial fibrosis (42). An in vitro model of HG-induced cardiac injury was established in the present study using the H9c2 cell line. In this model, cardiomyocytes were cultured in a medium containing high concentrations of glucose, similar to a long-term HG environment in vivo, which causes damage to cardiomyocyte-oxidative stress and cell apoptosis. In this case, the addition of various concentrations of Scu to the cell culture medium in advance protected cardiomyocytes by reducing oxidative stress and cell apoptosis, which is equivalent to preventing DCM.

The present study indicated that Scu could restore the number and morphology of H9c2 cells induced by HG, improve cell proliferation, and reduce ROS production and the apoptotic rate. Subsequent investigation indicated that Scu could reduce the protein expression levels of caspase-3, cleaved caspase-3, caspase-9, cleaved caspase-9, caspase-12, Bax, Nox2 and Nox4 in the HG-induced cardiac injury group (P<0.05); in addition, the protein expression levels of Bcl-2 were increased in H9c2 cells treated with Scu (P<0.05).

In conclusion, the present study demonstrated that Scu can regulate apoptosis and oxidative stress of H9c2 cells by modulating the protein expression levels of certain apoptotic factors and Nox in cells to alleviate the HG-induced cardiac injury and exert a protective effect on cardiomyocytes. The ROS levels were lower in the treated group than in the untreated group, suggesting that Scu may reduce ROS levels by directly scavenging free radicals or by promoting endogenous antioxidant enzymes while inhibiting apoptosis pathways, which work together to reduce intracellular oxidative stress and alleviate cell damage. This result has positive implications for further research into myocardial injury in DCM and clinical treatment of DCM, and it is also hoped that Scu can be used as an effective drug to prevent diabetic cardiomyopathy. However, the present study has some limitations, for example, it lacks in vivo experiments, which will be performed in future. The present study also lacks studies on molecular mechanisms and co-treatment of Scu with Nox activators or Nox overexpression plasmids, which is a target for future research.

In short, Scu is a bioactive compound mainly extracted from Erigeron breviscapus and has a variety of biological activities, including antioxidant and anti-apoptotic effects. Scu can directly scavenge ROS and other free radicals in the body, while upregulating the expression and activity of endogenous antioxidant enzymes, enhancing the antioxidant capacity of cells and reducing the level of oxidative stress. This helps protect cells from oxidative damage and reduces oxidative stress within cells. Scu can affect the expression and activity of proteins of the caspase family, inhibiting the activation of caspase-3 and caspase-9, thereby reducing cell apoptosis. This mechanism may be closely related to the antioxidant effect of Scu, as reducing oxidative stress may inhibit the activation of apoptosis signaling pathways. Scu can also alter the intracellular Bcl-2/Bax ratio by upregulating the anti-apoptotic protein Bcl-2 and downregulating the pro-apoptotic protein Bax, thus inhibiting the apoptotic signaling of the mitochondrial pathway. Scu affects oxidative stress and apoptosis through a variety of mechanisms, and the combined action of these mechanisms makes scutellarin potentially useful in cell protection and anti-oxidative damage.

Acknowledgements

The authors wish to thank Professor Yicheng Ma from Yunnan University (Kunming, China) for guidance with the experiments, Dr Haoan Yi from Kunming Medical University (Kunming, China) for guidance on both the experimental procedures and the structure of the article, and Dr Kenneth Otieno Onditi from Kunming Institute of Zoology, Chinese Academy of Sciences (Kunming, China) for helping with the language editing of the article.

Funding

Funding: The present study was supported by the National Natural Science Foundation of China (grant no. 82160533), and the Applied Basic Research Joint Special Fund Project of Science and Technology of the Department of Yunnan Province and Kunming Medical University (grant no. 2019FE001-016).

Availability of data and materials

The data generated in the present study may be requested from the corresponding author.

Authors' contributions

XW and ZL conceived and designed the study, conducted the formal analysis, interpreted the data, and wrote the original draft. ShL, BF, JY and SH conceived and designed the methodology, as well as reviewed and edited the manuscript. SiL conceived the study, interpreted the data, reviewed the manuscript and supervised the study. XW and SL confirm the authenticity of all the raw data. All authors read and approved the final manuscript.

Ethics approval and consent to participate

Not applicable.

Patient consent for publication

Not applicable.

Competing of interests

The authors declare that they have no competing interests.

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Copy and paste a formatted citation
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Spandidos Publications style
Wei X, Li Z, Li S, Yang J, Fan B, He S and Li S: &nbsp;Protective effect of scutellarin on myocardial cells treated with high glucose. Biomed Rep 23: 143, 2025.
APA
Wei, X., Li, Z., Li, S., Yang, J., Fan, B., He, S., & Li, S. (2025). &nbsp;Protective effect of scutellarin on myocardial cells treated with high glucose. Biomedical Reports, 23, 143. https://doi.org/10.3892/br.2025.2021
MLA
Wei, X., Li, Z., Li, S., Yang, J., Fan, B., He, S., Li, S."&nbsp;Protective effect of scutellarin on myocardial cells treated with high glucose". Biomedical Reports 23.3 (2025): 143.
Chicago
Wei, X., Li, Z., Li, S., Yang, J., Fan, B., He, S., Li, S."&nbsp;Protective effect of scutellarin on myocardial cells treated with high glucose". Biomedical Reports 23, no. 3 (2025): 143. https://doi.org/10.3892/br.2025.2021