Dingxin Recipe(定心方) Prevents Ischemia/Reperfusion-Induced Arrhythmias via Up-regulating Prohibitin and Suppressing Inflammatory Responses

Objective： To identify the underlying mechanisms of the protective effects of Dingxin Recipe （定心方, DXR）, a Chinese compound prescription that has been used clinically in China for more than 20 years, on ischemia/reperfusion （I/R）-induced arrhythmias in rat model. Methods： A total of 30 rats were randomly divided into three groups： sham group, I/R group, and DXR-pretreated I/R （DXR-I/R） group. Rats in the DXR- I/R group were intragastrically administrated with DXR （12.5 g/kg per day） for consecutive 7 days, while rats in the sham and I/R groups were administrated with normal saline. Arrhythmias were introduced by I/R and electrocardiograms （ECG） were recorded. Two-dimensional （2-D） polyacrylamide gel electrophoresis and matrix- assisted laser desorption ionization time-of-flight mass spectrometry （MALDI-TOF MS） were used to identify differentially expressed proteins. Immunohistochemistry, real-time quantitative polymerase chain reaction （RQ- PCR）, Western blot, and enzyme-linked immunosorbent assay （ELISA） were performed to analyze proteins obtained in the above experiments. Results： DXR significantly reduced the incidence and mean duration of ventricular tachycardia and ventricular fibrillation and dramatically decreased the mortality, as well as arrhythmia score, compared with those of the I/R group. Among successfully identified proteins, prohibitin （PHB） and heart fatty acid binding protein （hFABP） were up-regulated in DXR-pretreated I/R rats compared with those of the I/R rats. In addition, compared with the I/R group, the level of glutathione （GSH） was elevated accompanied by reduced expressions of interleukin-6 （IL-6） and neutrophil infiltration in I/R rats with DXR pretreatment. Conclusions： DXR could alleviate I/R-induced arrhythmias, which might be related to increased expression of PHB. The enhanced expression of PHB prevented against the depletion of GSH and consequently inhibited apoptosis of cardiomyocytes. Furthermore, up-regulation of PHB might ameliorate I/R-induced cell death and leakage of hFABP by suppressing neutrophil infiltration and IL-6 expressions.

Dingxin recipe alleviates atherosclerosis injury in Apo E-knockout mice via downregulation of visfatin expression and inhibition of the visfatin-induced inflammatory response

OBJECTIVE:To further elucidate the mechanism underlying the anti-atherosclerotic effect of Dingxin recipe(DXR).METHODS:Fifty 6-week-old male Apo E^-/-mice were randomly divided into the following groups:model,simvastatin(5 mg·kg^-1·d^-1),DXR low-dose(9.30 g·kg^-1·d^-1),DXR middle-dose(18.59 g·kg^-1·d^-1)and DXR high-dose(37.18 g·kg^-1·d^-1)(n=10).Ten male C57BL/6J mice were used as the control group.All Apo E^-/-mice were fed a high-fat diet(HFD)and the control mice received a common diet.After HFD for 12 weeks,the mice were treated with DXR or simvastatin for another 12 weeks.The expression of inflammatory cytokines and visfatin was determined in serum and atherosclerotic lesions by enzyme-linked immunosorbent assay.Visfatin expression was also assessed in aortic atherosclerotic plaques.Cultured vessel endothelial cells(VECs)were pretreated with DXR sera prior to visfatin.The effects of DXR were analyzed to elucidate its protective mechanism against visfatin-induced inflammation in VECs.RESULTS:DXR regulated blood lipids and reduced tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),intercellular adhesion molecules-1(ICAM-1),vascular cell adhesion molecule-1(VCAM-1)and visfatin expression in Apo E^-/-mice,particularly at the higher doses.The areas of atherosclerotic lesions in the DXR groups were significantly smaller than those in the model group.DXR alleviated visfatin-induced VEC injury via downregulation of TNF-α,IL-6,ICAM-1 and VCAM-1 through mitogen-activated protein kinase pathways.CONCLUSION:DXR alleviated atherosclerosis injury via downregulation of visfatin expression and inhibition of the visfatin-induced inflammatory response in VECs.