Journal: TH Open: Companion Journal to Thrombosis and Haemostasis

Article Title: Induction of Diabetes Abolishes the Antithrombotic Effect of Clopidogrel in Apolipoprotein E–Deficient Mice

doi: 10.1055/s-0037-1605361

Figure Lengend Snippet: Effects of clopidogrel on blood physiological parameters in wild-type, apoE-deficient, and diabetic apoE-deficient mice

Article Snippet: Clopidogrel bisulfate (clopidogrel) was purchased from Kemprotec Limited (Cumbria, United Kingdom).

Techniques:

Journal: TH Open: Companion Journal to Thrombosis and Haemostasis

Article Title: Induction of Diabetes Abolishes the Antithrombotic Effect of Clopidogrel in Apolipoprotein E–Deficient Mice

doi: 10.1055/s-0037-1605361

Figure Lengend Snippet: Effects of clopidogrel on the patency of carotid arteries. Clopidogrel (10 mg/kg/d, orally) or vehicle was administered once daily for 5 days. Carotid artery occlusion was initiated by application of FeCl 3 and blood flow was determined for 1 hour. Individual patency data are shown for vehicle-treated WT (1), clopidogrel-treated WT (2), vehicle-treated apoE-deficient (3), clopidogrel-treated apoE-deficient (4), vehicle-treated diabetic apoE-deficient (5), and clopidogrel-treated diabetic apoE-deficient groups (6). Open and filled bars indicate patent and occluded arteries, respectively. The number on the left side of each column represents animal ID number. apoE, apolipoprotein E; WT, wild-type.

Article Snippet: Clopidogrel bisulfate (clopidogrel) was purchased from Kemprotec Limited (Cumbria, United Kingdom).

Techniques:

Journal: TH Open: Companion Journal to Thrombosis and Haemostasis

Article Title: Induction of Diabetes Abolishes the Antithrombotic Effect of Clopidogrel in Apolipoprotein E–Deficient Mice

doi: 10.1055/s-0037-1605361

Figure Lengend Snippet: Effect of clopidogrel on time to first occlusion. Clopidogrel (C) at 10 mg/kg/d (orally) or vehicle (V) was administered once daily for 5 days to wild-type (WT), apoE-deficient, and diabetic apoE-deficient mice ( n = 10 per group). Carotid artery occlusion was initiated by application of FeCl 3 2 hours after the last dose of clopidogrel, arterial blood flow was monitored, and the time in seconds to first occlusion was determined. The results are presented as the mean + SE ( n = 10). *** p < 0.001 versus each vehicle group, † p < 0.05 versus the vehicle group of WT mice, # p < 0.05 (Student's t -test).

Article Snippet: Clopidogrel bisulfate (clopidogrel) was purchased from Kemprotec Limited (Cumbria, United Kingdom).

Techniques:

Journal: TH Open: Companion Journal to Thrombosis and Haemostasis

Article Title: Induction of Diabetes Abolishes the Antithrombotic Effect of Clopidogrel in Apolipoprotein E–Deficient Mice

doi: 10.1055/s-0037-1605361

Figure Lengend Snippet: Hematoxylin–eosin-stained cross-sections of injured carotid arteries. ( A ) Vehicle-treated wild-type (WT) (animal ID no. 105 in ); ( B ) clopidogrel-treated WT (no. 202); ( C ) vehicle-treated apoE-deficient (no. 308); ( D ) clopidogrel-treated apoE-deficient (no. 406); ( E ) vehicle-treated diabetic apoE-deficient (no. 501); and ( F ) clopidogrel-treated diabetic apoE-deficient mice (no. 607). Scale bar = 100 μm.

Article Snippet: Clopidogrel bisulfate (clopidogrel) was purchased from Kemprotec Limited (Cumbria, United Kingdom).

Techniques: Staining

Journal: TH Open: Companion Journal to Thrombosis and Haemostasis

Article Title: Induction of Diabetes Abolishes the Antithrombotic Effect of Clopidogrel in Apolipoprotein E–Deficient Mice

doi: 10.1055/s-0037-1605361

Figure Lengend Snippet: Effects of clopidogrel on morphometric parameters of injured carotid arteries, thrombus area ( A ) and lumen stenosis ( B ). Clopidogrel (C) at 10 mg/kg/d (orally) or vehicle (V) was administered once daily for 5 days to wild-type (WT), apoE-deficient, and diabetic apoE-deficient mice ( n = 10 per group) and arterial occlusion was induced 2 hours after the last dose of clopidogrel. After completing blood flow monitoring, mice were euthanized. The FeCl 3 -injured regions of the carotid arteries were collected, further fixed in 10% neutral-buffered formalin and paraffin sections prepared, and stained. The lumen and thrombotic areas were measured using an image analysis software (WinROOF 2013), and lumen stenosis (100 × thrombus area/lumen area) was calculated. Results are presented as the mean + SE ( n = 10). *** p < 0.001 versus each vehicle group, ### p < 0.001 (Student's t -test).

Article Snippet: Clopidogrel bisulfate (clopidogrel) was purchased from Kemprotec Limited (Cumbria, United Kingdom).

Techniques: Staining, Software

Journal: TH Open: Companion Journal to Thrombosis and Haemostasis

Article Title: Induction of Diabetes Abolishes the Antithrombotic Effect of Clopidogrel in Apolipoprotein E–Deficient Mice

doi: 10.1055/s-0037-1605361

Figure Lengend Snippet: Effects of clopidogrel on platelet expression of ADP-induced activated GPIIb/IIIa expression. Clopidogrel (10 mg/kg/d) or vehicle was administered orally once daily for 5 days to wild-type (WT), apoE-deficient, and streptozotocin-induced diabetic apoE-deficient mice, and blood collected 2 hours after the last dose. Ex vivo expression of activated GPIIb/IIIa on platelets induced by 5 and 20 μM ADP was assessed using a flow cytometer. In the vehicle groups, ADP stimulation resulted in a significant increase in the expression of activated platelet GPIIb/IIIa compared with no agonist ( p < 0.01 for diabetic apoE-deficient mice, p < 0.001 for other groups). The results are presented as mean ± SE ( n = 10). ** p < 0.01 versus corresponding vehicle group (Student's t -test).

Article Snippet: Clopidogrel bisulfate (clopidogrel) was purchased from Kemprotec Limited (Cumbria, United Kingdom).

Techniques: Expressing, Ex Vivo, Flow Cytometry

Journal: TH Open: Companion Journal to Thrombosis and Haemostasis

Article Title: Induction of Diabetes Abolishes the Antithrombotic Effect of Clopidogrel in Apolipoprotein E–Deficient Mice

doi: 10.1055/s-0037-1605361

Figure Lengend Snippet: Effects of clopidogrel on PAR4 TRAP-induced expression of platelet P-selectin. Clopidogrel (10 mg/kg/d) or vehicle was administered orally once daily for 5 days to wild-type (WT), apoE-deficient, and diabetic apoE-deficient mice, and blood collected 2 hours after the last dose. Ex vivo expression of P-selectin on platelets induced by 100 and 300 µM PAR4 TRAP was assessed using a flow cytometer. In the vehicle groups, PAR4 TRAP stimulation resulted in a significant increase in platelet P-selectin expression compared with no agonist ( p < 0.001, all groups). The results are presented as the mean ± SE ( n = 10). * p < 0.05, ** p < 0.01 versus corresponding vehicle group; # p < 0.05 versus the vehicle group of diabetic apoE-deficient mice; †† p < 0.01 versus clopidogrel group of diabetic apoE-deficient mice (Student's t -test).

Article Snippet: Clopidogrel bisulfate (clopidogrel) was purchased from Kemprotec Limited (Cumbria, United Kingdom).

Techniques: Expressing, Ex Vivo, Flow Cytometry, Mouse Assay

Journal: Scientific Reports

Article Title: Indobufen alleviates apoptosis by the PI3K/Akt/eNOS pathway in myocardial ischemia‒reperfusion (I/R) injury

doi: 10.1038/s41598-025-17345-y

Figure Lengend Snippet: Indobufen alleviated myocardial ischemia injury in I/R rats. ( A )TTC staining of representative sections, the white-colored areas represent the infarct areas and the red-colored areas represent non-infarcted areas. ( B ) Quantification of rat heart infarct size in different groups. Lane1: I/R group, lane2: clopidogrel (7.5 mg/kg) group, lane3: indobufen (20 mg/kg) group, lane4: indobufen (10 mg/kg) group, lane5: aspirin (10 mg/kg), lane6: aspirin (5 mg/kg) group. ( C ) Quantification of pathological indicators in H&E staining: hyperemia, hemorrhage, necrosis and degeneration. The detailed scoring criterion of pathological indicators was mentioned above. Lane1: sham group, lane2: I/R group, lane3: clopidogrel (7.5 mg/kg) group, lane4: indobufen (20 mg/kg) group, lane5: indobufen (10 mg/kg) group, lane6: aspirin (10 mg/kg), lane7: aspirin (5 mg/kg) group. ( D ) Histological analysis showed cardioprotection of Indubufen in I/R hearts (400×). Scale bar is 25 μm. ( E ) The arrow points to the observation site. All data are shown as mean ± S.E.M, n = 7-8. * P < 0.05, ** P < 0.01 vs. I/R group; # P < 0.05, ## P < 0.01 vs. Sham.

Article Snippet: Clopidogrel hydrogen sulfate tablets (Lot 7A356, with each tablet containing 75 mg of active ingredients) were purchased from Sanofi ClirSNC (Cat. #7A356, Sanofi Winthrop Industrie, France).

Techniques: Staining

Journal: Scientific Reports

Article Title: Indobufen alleviates apoptosis by the PI3K/Akt/eNOS pathway in myocardial ischemia‒reperfusion (I/R) injury

doi: 10.1038/s41598-025-17345-y

Figure Lengend Snippet: Indobufen improved left ventricular function in I/R rats. ( A ) Representative images of each group of echocardiograms. ( B ) Comparison of EF (%) in each group. ( C ) comparison of FS (%) in each group. All data are shown as mean ± S.E.M, n = 5. ( B , C ) Lane1: sham group, lane2: I/R group, lane3: clopidogrel (7.5 mg/kg) group, lane4: indobufen (20 mg/kg) group, lane5: indobufen (10 mg/kg) group, lane6: aspirin (10 mg/kg), lane7: aspirin (5 mg/kg) group. * P < 0.05, ** P < 0.01 vs. I/R group; # P < 0.05, ## P < 0.01 vs. Sham.

Article Snippet: Clopidogrel hydrogen sulfate tablets (Lot 7A356, with each tablet containing 75 mg of active ingredients) were purchased from Sanofi ClirSNC (Cat. #7A356, Sanofi Winthrop Industrie, France).

Techniques: Comparison

Journal: Scientific Reports

Article Title: Indobufen alleviates apoptosis by the PI3K/Akt/eNOS pathway in myocardial ischemia‒reperfusion (I/R) injury

doi: 10.1038/s41598-025-17345-y

Figure Lengend Snippet: Indobufen alleviated oxidative stress injury in I/R rats. ( A ) Indobufen suppressed the up-regulation of CK level of I/R group. ( B ) Indobufen decreased LDH activity of I/R group. ( C ) Indobufen reduced MDA level of I/R group. ( D ) Indobufen up-regulated SOD level of I/R group. ( A – D ) Lane1: sham group, lane2: I/R group, lane3: clopidogrel (7.5 mg/kg) group, lane4: indobufen (20 mg/kg) group, lane5: indobufen (10 mg/kg) group, lane6: aspirin (10 mg/kg), lane7: aspirin (5 mg/kg) group. Data are shown as mean ± S.E.M, n = 8. * P < 0.05, ** P < 0.01 vs. I/R group; # P < 0.05, ## P < 0.01 vs. Sham.

Article Snippet: Clopidogrel hydrogen sulfate tablets (Lot 7A356, with each tablet containing 75 mg of active ingredients) were purchased from Sanofi ClirSNC (Cat. #7A356, Sanofi Winthrop Industrie, France).

Techniques: Activity Assay

Journal: Scientific Reports

Article Title: Indobufen alleviates apoptosis by the PI3K/Akt/eNOS pathway in myocardial ischemia‒reperfusion (I/R) injury

doi: 10.1038/s41598-025-17345-y

Figure Lengend Snippet: Indobufen significantly elevated plasma levels of NO and eNOS and cardiac mRNA levels of eNOS, and down-regulated plasma ROS levels. ( A ) Indobufen elevated the level of eNOS protein in plasma ( B ) Indobufen up-regulated the level of eNOS mRNA in heart. ( C ) Indobufen increased NO production in rat plasma. ( D ) Indobufen reduced ROS generation in rat plasma. Lane1: sham group, lane2: I/R group, lane3: clopidogrel (7.5 mg/kg) group, lane4: indobufen (20 mg/kg) group, lane5: indobufen (10 mg/kg) group, lane6: aspirin (10 mg/kg), lane7: aspirin (5 mg/kg) group. All data are shown as mean ± S.E.M, n = 5–8. * P < 0.05, ** P < 0.01 vs. I/R group; # P < 0.05, ## P < 0.01 vs. Sham.

Article Snippet: Clopidogrel hydrogen sulfate tablets (Lot 7A356, with each tablet containing 75 mg of active ingredients) were purchased from Sanofi ClirSNC (Cat. #7A356, Sanofi Winthrop Industrie, France).

Techniques: Clinical Proteomics

Journal: Scientific Reports

Article Title: Indobufen alleviates apoptosis by the PI3K/Akt/eNOS pathway in myocardial ischemia‒reperfusion (I/R) injury

doi: 10.1038/s41598-025-17345-y

Figure Lengend Snippet: Indobufen significantly inhibited cardiomyocyte apoptosis. ( A ) Suppression of cardiomyocyte apoptosis by Indobufen pretreatment (×400). Scale bar is 25 μm. Brown staining of the nucleus is indicative of apoptosis. The arrow points to the observation site ( B ) Percentage of apoptotic cells, n = 5. ( C ) Caspase-3 protein level was determined using Western blot analysis. ( D ) Quantification of Caspase-3 protein expression, n = 3. ( E ) Bax and BCL-2 protein levels were determined using Western blot analysis. ( F ) Quantification of Bax and BCL-2 proteins expression, n = 3. Each column represents the mean ± S.E.M. ( B , D , F ) Lane1: sham group, lane2: I/R group, lane3: clopidogrel (7.5 mg/kg) group, lane4: indobufen (20 mg/kg) group, lane5: indobufen (10 mg/kg) group, lane6: aspirin (10 mg/kg), lane7: aspirin (5 mg/kg) group. * P < 0.05, ** P < 0.01 vs. I/R group; # P < 0.05, ## P < 0.01 vs. Sham. Original blots/gels are presented in Supplementary.

Article Snippet: Clopidogrel hydrogen sulfate tablets (Lot 7A356, with each tablet containing 75 mg of active ingredients) were purchased from Sanofi ClirSNC (Cat. #7A356, Sanofi Winthrop Industrie, France).

Techniques: Staining, Western Blot, Expressing

Journal: Scientific Reports

Article Title: Indobufen alleviates apoptosis by the PI3K/Akt/eNOS pathway in myocardial ischemia‒reperfusion (I/R) injury

doi: 10.1038/s41598-025-17345-y

Figure Lengend Snippet: Effects of Indobufen on the expression of p-Akt and p-eNOS in cardiac tissue. Representative photomicrographs of immunohistochemical staining showed the effects of Indobufen on the expression of ( A ) p-Akt and ( B ) p-eNOS immunoreactivity in heart sections. All the photographs were taken at 400×. Scale bar = 25 μm. ( C ) The positive staining of p-Akt (%area). ( D ) The positive staining of p-eNOS (% area). ( E ) p-Akt/Akt, p-eNOS/eNOS protein levels were determined using western blot analysis. ( F ) Quantification of p-Akt/Akt proteins expression. ( G ) Quantification of p-eNOS/eNOS proteins expression. Lane1: sham group, lane2: I/R group, lane3: clopidogrel (7.5 mg/kg) group, lane4: indobufen (20 mg/kg) group, lane5: indobufen (10 mg/kg) group, lane6: aspirin (10 mg/kg), lane7: aspirin (5 mg/kg) group. Each column represents the mean ± S.E.M, n = 5. * P < 0.05, ** P < 0.01 vs. I/R group; # P < 0.05, ## P < 0.01 vs. Sham. Original blots/gels are presented in Supplementary.

Article Snippet: Clopidogrel hydrogen sulfate tablets (Lot 7A356, with each tablet containing 75 mg of active ingredients) were purchased from Sanofi ClirSNC (Cat. #7A356, Sanofi Winthrop Industrie, France).

Techniques: Expressing, Immunohistochemical staining, Staining, Western Blot