ABSTRACT
ObjectiveTo reveal the effect of Wenxin prescription on mitochondrial energy metabolism and silent information regulator 1 （SIRT1）/peroxisome proliferator-activated receptor-γ coactivator-1α （PGC-1α）/recombinant estrogen-related receptor α （ERRα） signaling pathway in rats with myocardial ischemia-reperfusion injury. MethodTotally 90 male Wistar rats of SPF grade were randomly assigned into a sham operation group， a model group， and low-， medium-， and high-dose Wenxin prescription groups， with 18 rats in each group. The rats in low-， medium-， and high-dose Wenxin prescription groups were administrated with 0.99， 1.98， and 3.96 g·kg-1 granules by gavage， respectively， and those in the sham operation group and model group with the same amount of normal saline. Twenty-one days after pre-administration， the rat model of myocardial ischemia-reperfusion injury was established by ligation of the left anterior descending coronary artery for 30 min and reperfusion for 2 h， and the rats in the sham operation group were only threaded without ligation. Myocardial infarction area was observed through 2，3，5-triphenyl-2h-tetrazolium chloride （TTC） staining， and the myocardial histopathology through hematoxylin-eosin （HE） staining. The levels of creatine kinase-MB （CK-MB） and lactate dehydrogenase （LDH） in serum， cytochrome C oxidase （CCO） and succinate dehydrogenase （SDH） in mitochondrion， and ATP in myocardial tissue were detected according to kit instructions. The mRNA and protein levels of SIRT1， PGC-1α， ERRα， and mitochondrial transcription factor A （TFAM） in myocardial tissue were determined by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. ResultCompared with the sham operation group， the model group showed broken and disordered myocardial fibers， cytoplasmic edema， and pyknosis and deviation of nuclei. Moreover， the modeling increased the levels of CK-MB and LDH （P<0.05， P<0.01）， lowered the levels of ATP， CCO， and SDH （P<0.05， P<0.01）， and down-regulated the mRNA and protein levels of SIRT1， PGC-1α， ERRα， and TFAM in myocardial tissue （P<0.05， P<0.01）. Compared with the model group， Wenxin prescription reduced the myocardial infarction area （especially in the high-dose group， P<0.01）， restored the pathological changes， lowered the levels of CK-MB and LDH （P<0.05， P<0.01）， increased the levels of ATP， CCO， and SDH （especially in the high-dose group， P<0.01）， and up-regulated the mRNA and protein levels of SIRT1， PGC-1α， ERRα， and TFAM in myocardial tissue （P<0.05， P<0.01）. ConclusionWenxin prescription can protect rats from myocardial ischemia-reperfusion injury by regulating myocardial mitochondrial energy metabolism via the SIRT1/PGC-1α/ERRα signaling pathway.
ABSTRACT
Objective:To observe the effects of Wenxin prescription on the key targets of gap 1/synthesis （G<sub>1</sub>/S） cell cycle transformation in rats with atherosclerosis （AS）， and reveal the mechanism of Wenxin prescription in the treatment of AS. Method:Ninety SPF Wistar rats were randomly divided into a normal group （<italic>n</italic>=6） and a modeling group （<italic>n</italic>=84）. The rats in the modeling group were fed on a high-fat diet （4% cholesterol， 0.5% sodium cholate， 0.2% propyl thiouracil， 10% lard， 5% sugar， and 80.3% basal feed） for 60 days， and intraperitoneally injected with 400 000 U·kg<sup>-1 </sup>vitamin D<sub>3</sub>， once a week for three weeks. The model rats were then randomly divided into a model group， high-dose （24 g·kg<sup>-1</sup>）， medium-dose （12 g·kg<sup>-1</sup>）， and low-dose （6 g·kg<sup>-1</sup>） Wenxin prescription groups， and a rosuvastatin （1.8 mg·kg<sup>-1</sup>） group. The groups with drug intervention were treated correspondingly by gavage for 30 days. The rats in the model group were administered with an equal volume of distilled water. The general condition of rats was observed after treatment. The levels of high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， and total cholesterol （CHO） were detected by enzyme-linked immunosorbent assay （ELISA）， and the atherosclerosis index （AI） was calculated. The pathological morphology of the coronary artery and aorta was observed by hematoxylin-eosin （HE） staining. The protein and mRNA expression of E2F transcription factor 1 （E2F1）， phosphorylated retinoblastoma protein （p-Rb）， cell division cycle 25 （Cdc25）， CyclinE， and CyclinD<sub>1</sub> was detected by Western blot and real-time fluorescence-based quantitative polymerase chain reaction （Real-time-PCR）， respectively. Result:Compared with the normal group， the model group showed intima thickening， smooth muscle proliferation， and plaque formation in the coronary artery and aorta， decreased HDL-C （<italic>P</italic><0.01）， increased LDL-C， CHO， and AI （<italic>P</italic><0.01）， elevated protein and mRNA expression of E2F1， Cdc25， p-Rb， CyclinE and CyclinD<sub>1</sub> （<italic>P</italic><0.05）. Compared with the model group， the rosuvastatin group and the Wenxin prescription groups showed slight intimal hyperplasia and lumen narrowing of the coronary artery and aorta， decreased levels of LDL-C， CHO， and AI （<italic>P</italic><0.01）， and declining protein and mRNA expression of E2F1， Cdc25， p-Rb， CyclinE， and CyclinD<sub>1</sub> to varying degrees （<italic>P</italic><0.05）. Conclusion:Wenxin prescription can significantly inhibit the expression of key proteins and genes of the G<sub>1</sub>/S cell cycle， regulate G<sub>1</sub>/S cell cycle transformation， and reduce vascular smooth muscle and intimal hyperplasia in AS rats.
