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Protective Effects of Kaempferol against Myocardial Ischemia/Reperfusion Injury in Isolated Rat Heart via Antioxidant Activity and Inhibition of Glycogen Synthase Kinase-3β.

Zhou M, Ren H, Han J, Wang W, Zheng Q, Wang D - Oxid Med Cell Longev (2015)

Bottom Line: Moreover, total glycogen synthase kinase-3β (GSK-3β), phospho-GSK-3β (P-GSK-3β), precaspase-3, cleaved caspase-3, and cytoplasm cytochrome C were assayed using Western blot analysis.Pretreatment with kaempferol significantly improved the recovery of LVDP and ±dp/dt max, as well as increased the levels of SOD and P-GSK-3β and GSH/GSSG ratio.However, the pretreatment reduced myocardial infarct size and TUNEL-positive cell rate, as well as decreased the levels of cleaved caspase-3, cytoplasm cytochrome C, CK, LDH, MDA, and TNF-α.

View Article: PubMed Central - PubMed

Affiliation: Shandong Provincial Qianfoshan Hospital, Jinan 250014, China ; Weifang Medical University, Weifang 261031, China.

ABSTRACT

Objective: This study aimed to evaluate the protective effect of kaempferol against myocardial ischemia/reperfusion (I/R) injury in rats.

Method: Left ventricular developed pressure (LVDP) and its maximum up/down rate (±dp/dt max) were recorded as myocardial function. Infarct size was detected with 2,3,5-triphenyltetrazolium chloride staining. Cardiomyocyte apoptosis was determined using terminal deoxynucleotidyl nick-end labeling (TUNEL). The levels of creatine kinase (CK), lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione/glutathione disulfide (GSH/GSSG) ratio, and tumor necrosis factor-alpha (TNF-α) were determined using enzyme linked immunosorbent assay (ELISA). Moreover, total glycogen synthase kinase-3β (GSK-3β), phospho-GSK-3β (P-GSK-3β), precaspase-3, cleaved caspase-3, and cytoplasm cytochrome C were assayed using Western blot analysis.

Results: Pretreatment with kaempferol significantly improved the recovery of LVDP and ±dp/dt max, as well as increased the levels of SOD and P-GSK-3β and GSH/GSSG ratio. However, the pretreatment reduced myocardial infarct size and TUNEL-positive cell rate, as well as decreased the levels of cleaved caspase-3, cytoplasm cytochrome C, CK, LDH, MDA, and TNF-α.

Conclusion: These results suggested that kaempferol provides cardioprotection via antioxidant activity and inhibition of GSK-3β activity in rats with I/R.

No MeSH data available.


Related in: MedlinePlus

Kaempferol prevents myocardial cell apoptosis in I/R. (a) Cell apoptosis was analyzed using TUNEL staining. Magnification: 400x. (b) Apoptosis cells/total cells (%). Values are presented as means with their standard deviations (, n = 8); ∗∗P < 0.01, compared with the control group; ##P < 0.01 and #P < 0.05, compared with the I/R group.
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fig4: Kaempferol prevents myocardial cell apoptosis in I/R. (a) Cell apoptosis was analyzed using TUNEL staining. Magnification: 400x. (b) Apoptosis cells/total cells (%). Values are presented as means with their standard deviations (, n = 8); ∗∗P < 0.01, compared with the control group; ##P < 0.01 and #P < 0.05, compared with the I/R group.

Mentions: TUNEL assay was carried out according to the manufacturer's instructions. After deparaffinization and rehydration, the sections were treated with 10 mmol/L protease K for 15 min. The slides were immersed in TUNEL reaction mixture for 60 min at 37°C in a humidified atmosphere in the dark. A converter POD was used to incubate the slides for 30 min. The slides were then analyzed using an optical microscope. To evaluate the apoptosis index of the TUNEL-stained heart tissues, we captured 10 random fields per tissue section at 400x magnification. TUNEL index (%) is calculated as the ratio of the number of TUNEL-positive cells divided by the total number of cells [20] (see Figure 4).


Protective Effects of Kaempferol against Myocardial Ischemia/Reperfusion Injury in Isolated Rat Heart via Antioxidant Activity and Inhibition of Glycogen Synthase Kinase-3β.

Zhou M, Ren H, Han J, Wang W, Zheng Q, Wang D - Oxid Med Cell Longev (2015)

Kaempferol prevents myocardial cell apoptosis in I/R. (a) Cell apoptosis was analyzed using TUNEL staining. Magnification: 400x. (b) Apoptosis cells/total cells (%). Values are presented as means with their standard deviations (, n = 8); ∗∗P < 0.01, compared with the control group; ##P < 0.01 and #P < 0.05, compared with the I/R group.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4525766&req=5

fig4: Kaempferol prevents myocardial cell apoptosis in I/R. (a) Cell apoptosis was analyzed using TUNEL staining. Magnification: 400x. (b) Apoptosis cells/total cells (%). Values are presented as means with their standard deviations (, n = 8); ∗∗P < 0.01, compared with the control group; ##P < 0.01 and #P < 0.05, compared with the I/R group.
Mentions: TUNEL assay was carried out according to the manufacturer's instructions. After deparaffinization and rehydration, the sections were treated with 10 mmol/L protease K for 15 min. The slides were immersed in TUNEL reaction mixture for 60 min at 37°C in a humidified atmosphere in the dark. A converter POD was used to incubate the slides for 30 min. The slides were then analyzed using an optical microscope. To evaluate the apoptosis index of the TUNEL-stained heart tissues, we captured 10 random fields per tissue section at 400x magnification. TUNEL index (%) is calculated as the ratio of the number of TUNEL-positive cells divided by the total number of cells [20] (see Figure 4).

Bottom Line: Moreover, total glycogen synthase kinase-3β (GSK-3β), phospho-GSK-3β (P-GSK-3β), precaspase-3, cleaved caspase-3, and cytoplasm cytochrome C were assayed using Western blot analysis.Pretreatment with kaempferol significantly improved the recovery of LVDP and ±dp/dt max, as well as increased the levels of SOD and P-GSK-3β and GSH/GSSG ratio.However, the pretreatment reduced myocardial infarct size and TUNEL-positive cell rate, as well as decreased the levels of cleaved caspase-3, cytoplasm cytochrome C, CK, LDH, MDA, and TNF-α.

View Article: PubMed Central - PubMed

Affiliation: Shandong Provincial Qianfoshan Hospital, Jinan 250014, China ; Weifang Medical University, Weifang 261031, China.

ABSTRACT

Objective: This study aimed to evaluate the protective effect of kaempferol against myocardial ischemia/reperfusion (I/R) injury in rats.

Method: Left ventricular developed pressure (LVDP) and its maximum up/down rate (±dp/dt max) were recorded as myocardial function. Infarct size was detected with 2,3,5-triphenyltetrazolium chloride staining. Cardiomyocyte apoptosis was determined using terminal deoxynucleotidyl nick-end labeling (TUNEL). The levels of creatine kinase (CK), lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione/glutathione disulfide (GSH/GSSG) ratio, and tumor necrosis factor-alpha (TNF-α) were determined using enzyme linked immunosorbent assay (ELISA). Moreover, total glycogen synthase kinase-3β (GSK-3β), phospho-GSK-3β (P-GSK-3β), precaspase-3, cleaved caspase-3, and cytoplasm cytochrome C were assayed using Western blot analysis.

Results: Pretreatment with kaempferol significantly improved the recovery of LVDP and ±dp/dt max, as well as increased the levels of SOD and P-GSK-3β and GSH/GSSG ratio. However, the pretreatment reduced myocardial infarct size and TUNEL-positive cell rate, as well as decreased the levels of cleaved caspase-3, cytoplasm cytochrome C, CK, LDH, MDA, and TNF-α.

Conclusion: These results suggested that kaempferol provides cardioprotection via antioxidant activity and inhibition of GSK-3β activity in rats with I/R.

No MeSH data available.


Related in: MedlinePlus