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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 increases the phosphorylation of GSK-3β and reduces the release of cytochrome C and the dissociation of caspase-3. (a) Representative Western blots for T-GSK-3β and P-GSK-3β (Ser9). Lanes 1 and 2 are the control group, lanes 3 and 4 are the I/R group, lanes 5 and 6 are the kaempferol group, and lanes 7 and 8 are the TDZD-8 group. (b) Grey value analysis demonstrates that kaempferol increases the ratio of P-GSK-3β (ser)/T-GSK-3β. (c) Representative Western blot for cleaved caspase-3 and cytoplasm cytochrome C. Lane 1 is the control group, lane 2 is the I/R group, lane 3 is the kaempferol group, and lane 4 is the TDZD-8 group. (d) Grey value analysis demonstrates that kaempferol reduces the ratio of cleaved caspase-3/β-actin. (e) Grey value analysis demonstrates that kaempferol reduces the ratio of cytoplasm cytochrome C/β-actin. Values are presented as means with their standard deviations (, n = 8); ∗∗P < 0.01, compared with the control group; ##P < 0.01, compared with the I/R group.
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fig5: Kaempferol increases the phosphorylation of GSK-3β and reduces the release of cytochrome C and the dissociation of caspase-3. (a) Representative Western blots for T-GSK-3β and P-GSK-3β (Ser9). Lanes 1 and 2 are the control group, lanes 3 and 4 are the I/R group, lanes 5 and 6 are the kaempferol group, and lanes 7 and 8 are the TDZD-8 group. (b) Grey value analysis demonstrates that kaempferol increases the ratio of P-GSK-3β (ser)/T-GSK-3β. (c) Representative Western blot for cleaved caspase-3 and cytoplasm cytochrome C. Lane 1 is the control group, lane 2 is the I/R group, lane 3 is the kaempferol group, and lane 4 is the TDZD-8 group. (d) Grey value analysis demonstrates that kaempferol reduces the ratio of cleaved caspase-3/β-actin. (e) Grey value analysis demonstrates that kaempferol reduces the ratio of cytoplasm cytochrome C/β-actin. Values are presented as means with their standard deviations (, n = 8); ∗∗P < 0.01, compared with the control group; ##P < 0.01, compared with the I/R group.

Mentions: As shown in Figure 5, the level of GSK-3β phosphorylation in the kaempferol (0.65 ± 0.039) and TDZD-8 groups (0.74 ± 0.043) significantly increased compared with those in the control (0.32 ± 0.018) and I/R groups (0.35 ± 0.02) (P < 0.01). However, no evident difference was observed between the control and I/R groups. The release of cytochrome C and the dissociation of caspase-3 in the I/R groups (0.52 ± 0.039 and 0.37 ± 0.021, resp.) were significantly increased compared with those in the control groups (P < 0.01), whereas the kaempferol (0.32 ± 0.024 and 0.25 ± 0.018, resp.) and TDZD-8 groups (0.21 ± 0.013 and 0.29 ± 0.019, resp.) were significantly decreased compared with those in the I/R groups (P < 0.01).


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 increases the phosphorylation of GSK-3β and reduces the release of cytochrome C and the dissociation of caspase-3. (a) Representative Western blots for T-GSK-3β and P-GSK-3β (Ser9). Lanes 1 and 2 are the control group, lanes 3 and 4 are the I/R group, lanes 5 and 6 are the kaempferol group, and lanes 7 and 8 are the TDZD-8 group. (b) Grey value analysis demonstrates that kaempferol increases the ratio of P-GSK-3β (ser)/T-GSK-3β. (c) Representative Western blot for cleaved caspase-3 and cytoplasm cytochrome C. Lane 1 is the control group, lane 2 is the I/R group, lane 3 is the kaempferol group, and lane 4 is the TDZD-8 group. (d) Grey value analysis demonstrates that kaempferol reduces the ratio of cleaved caspase-3/β-actin. (e) Grey value analysis demonstrates that kaempferol reduces the ratio of cytoplasm cytochrome C/β-actin. Values are presented as means with their standard deviations (, n = 8); ∗∗P < 0.01, compared with the control group; ##P < 0.01, compared with the I/R group.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig5: Kaempferol increases the phosphorylation of GSK-3β and reduces the release of cytochrome C and the dissociation of caspase-3. (a) Representative Western blots for T-GSK-3β and P-GSK-3β (Ser9). Lanes 1 and 2 are the control group, lanes 3 and 4 are the I/R group, lanes 5 and 6 are the kaempferol group, and lanes 7 and 8 are the TDZD-8 group. (b) Grey value analysis demonstrates that kaempferol increases the ratio of P-GSK-3β (ser)/T-GSK-3β. (c) Representative Western blot for cleaved caspase-3 and cytoplasm cytochrome C. Lane 1 is the control group, lane 2 is the I/R group, lane 3 is the kaempferol group, and lane 4 is the TDZD-8 group. (d) Grey value analysis demonstrates that kaempferol reduces the ratio of cleaved caspase-3/β-actin. (e) Grey value analysis demonstrates that kaempferol reduces the ratio of cytoplasm cytochrome C/β-actin. Values are presented as means with their standard deviations (, n = 8); ∗∗P < 0.01, compared with the control group; ##P < 0.01, compared with the I/R group.
Mentions: As shown in Figure 5, the level of GSK-3β phosphorylation in the kaempferol (0.65 ± 0.039) and TDZD-8 groups (0.74 ± 0.043) significantly increased compared with those in the control (0.32 ± 0.018) and I/R groups (0.35 ± 0.02) (P < 0.01). However, no evident difference was observed between the control and I/R groups. The release of cytochrome C and the dissociation of caspase-3 in the I/R groups (0.52 ± 0.039 and 0.37 ± 0.021, resp.) were significantly increased compared with those in the control groups (P < 0.01), whereas the kaempferol (0.32 ± 0.024 and 0.25 ± 0.018, resp.) and TDZD-8 groups (0.21 ± 0.013 and 0.29 ± 0.019, resp.) were significantly decreased compared with those in the I/R groups (P < 0.01).

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