Rosuvastatin relieves myocardial ischemia/reperfusion injury by upregulating PPAR‑γ and UCP2

Figures

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  Figure 1.. RS increases SOD activity, and decreases LDH, CK-MB, MDA and troponin I/T activities. A total of 48 rabbits were randomly divided into four groups, including the sham group, MI/R group, RS group and RS + ATR group. (A) SOD activity was detected using a commercial kit. (B) LDH activity was measured using a cytotoxicity assay. (C) CK-MB activity was analyzed using a CK-MB assay. (D) MDA activity was analyzed using a MDA activity assay kit. (E) Troponin I/T activities were measured by ELISA. **P<0.01 and ***P<0.001 vs. the sham group; #P<0.05, ##P<0.01 and ###P<0.001 vs. the MI/R group; ++P<0.01 and +++P<0.001 vs. the RS group. RS, rosuvastatin; ATR, atractyloside; SOD, superoxide dismutase; LDH, lactate dehydrogenase; CK-MB, creatine kinase-muscle/brain, MDA, malondialdehyde; MI/R, myocardial ischemia/reperfusion.

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  Figure 2.. RS inhibits myocardial infarct size. A total of 48 rabbits were randomly divided into four groups, including the sham group, MI/R group, RS group and RS + ATR group. (A) Images of Evans blue/TTC-stained sections were captured. (B) The infarct size (% AN/AAR) was detected in four groups. ***P<0.001 vs. the sham group; ###P<0.001 vs. the MI/R group; ++P<0.01 vs. the RS group. MI/R, myocardial ischemia/reperfusion; RS, rosuvastatin; ATR, atractyloside; AN, area of necrosis; AAR, area-at-risk.

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  Figure 3.. RS downregulates caspase-9 and cyt c expression, and upregulates UCP2 and PPAR-γ expression. A total of 48 rabbits were randomly divided into four groups, including the sham group, MI/R group, RS group and RS + ATR group. mRNA and protein expression levels of caspase-9, cyt c, UCP2 and PPAR-γ were detected by (A) reverse transcription-quantitative polymerase chain reaction and (B) western blotting. (C) The results of western blotting were quantitatively analyzed. ***P<0.001 vs. the sham group; #P<0.05 and ###P<0.001 vs. the MI/R group; +P<0.05, ++P<0.01 and +++P<0.001 vs. the RS group. Cyt c, cytochrome c; UCP2, mitochondrial uncoupling protein 2; PPAR-γ, peroxisome proliferator-activated receptor-γ; MI/R, myocardial ischemia/reperfusion; RS, rosuvastatin; ATR, atractyloside.

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  Figure 4.. Pretreatment with RS promotes cardiomyocyte viability and inhibits LDH release following OGD/R damage. Cardiomyocytes were divided into the following groups: Control, control + siPPAR-γ, OGD/R, OGD/R + siPPAR-γ, RS + OGD/R, RS + OGD/R + siPPAR-γ, ATR + RS + OGD/R, ATR + RS + OGD/R + siPPAR-γ. (A) Following reperfusion, cell viability was evaluated by MTT assay. (B) LDH activity was measured using LDH cytotoxicity assay. *P<0.05 and ***P<0.001 vs. the control group; #P<0.05 and ###P<0.001 vs. the OGD/R group; +P<0.05 and ++P<0.01 vs. the OGD/R + siPPAR-γ group; OGD/R, oxygen-glucose deprivation/reperfusion; LDH, lactate dehydrogenase; OD, optical density; si, small interfering RNA; PPAR-γ, peroxisome proliferator-activated receptor-γ; RS, rosuvastatin; ATR, atractyloside.

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  Figure 5.. Pretreatment with RS reduces ROS production following OGD/R damage. Cardiomyocytes were divided into the following groups: Control, control + siPPAR-γ, OGD/R, OGD/R + siPPAR-γ, RS + OGD/R, RS + OGD/R + siPPAR-γ, ATR + RS + OGD/R, ATR + RS + OGD/R + siPPAR-γ. Flow cytometry was used to determine the (A) mitochondrial and (B) cellular ROS levels. M1 indicates the cells that emitted a DCF signal and therefore the level of ROS production. *P<0.05 and **P<0.01 vs. the control group; #P<0.05 and ##P<0.01 vs. the OGD/R group; +P<0.05 and ++P<0.01 vs. the OGD/R + siPPAR-γ group; −P<0.05 vs. the RS + OGD/R + siPPAR-γ group. OGD/R, oxygen-glucose deprivation/reperfusion; LDH, lactate dehydrogenase; DCF, 2′,7′-dichlorofluorescein; ROS, reactive oxygen species; RS, rosuvastatin; ATR, atractyloside; si, small interfering RNA; PPAR-γ, peroxisome proliferator-activated receptor-γ.

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  Figure 6.. Pretreatment with RS decreases caspase-9 and cyt c mRNA expression, and increases UCP2 and PPAR-γ mRNA expression following OGD/R damage. Cardiomyocytes were divided into the following groups: Control, control + siPPAR-γ, OGD/R, OGD/R + siPPAR-γ, RS + OGD/R, RS + OGD/R + siPPAR-γ, ATR + RS + OGD/R, ATR + RS + OGD/R + siPPAR-γ. Reverse transcription-quantitative polymerase chain reaction was performed to analyze the mRNA expression levels of (A) caspase-9, (B) cyt c, (C) UCP2 and (D) PPAR-γ in cardiomyocytes. *P<0.05 and ***P<0.001 vs. the control group; #P<0.05 and ##P<0.01 vs. the OGD/R group; +P<0.05 and ++P<0.01 vs. the OGD/R + siPPAR-γ group; −P<0.05 and −−P<0.01 vs. the RS + OGD/R + siPPAR-γ group. OGD/R, oxygen-glucose deprivation/reperfusion; cyt c, cytochrome c; RS, rosuvastatin; ATR, atractyloside; si, small interfering RNA; PPAR-γ, peroxisome proliferator-activated receptor-γ; UCP2, mitochondrial uncoupling protein 2.

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  Figure 7.. RS downregulates protein expression levels of caspase-9 and cyt c, and upregulates protein expression levels of UCP2 and PPAR-γ following OGD/R damage. Cardiomyocytes were divided into the following groups: Control, control + siPPAR-γ, OGD/R, OGD/R + siPPAR-γ, RS + OGD/R, RS + OGD/R + siPPAR-γ, ATR + RS + OGD/R, ATR + RS + OGD/R + siPPAR-γ. (A) Western blot analysis was used to detect the protein expression levels of caspase-9, cyt c, UCP2 and PPAR-γ in cardiomyocytes. (B) The protein expression levels were quantitatively analyzed according to the protein gray values. **P<0.01 and ***P<0.001 vs. the control group; #P<0.05 and ##P<0.01 vs. the OGD/R group; +P<0.05 and ++P<0.01 vs. the OGD/R + siPPAR-γ group; −−P<0.01 vs. the RS + OGD/R + siPPAR-γ group. OGD/R, oxygen-glucose deprivation/reperfusion; cyt c, cytochrome c; PPAR-γ, peroxisome proliferator-activated receptor-γ; UCP2, mitochondrial uncoupling protein 2; RS, rosuvastatin; ATR, atractyloside; si, small interfering RNA.