The Research Of Guanxinping On Endothelial Dysfunction And Signaling Pathway Disorder

Coronary artery disease refers to coronary atherosclerosis block blood vessel. Heart disease caused by myocardial referred to CHD. CHD is a serious hazard to human health, and a common diseases. In Europe and the United States and other developed countries, the mortality rate of CHD accounted for the first of the mortality of various diseases. According to statistics, CHD prevalence was6.49%in China. CHD morbidity and mortality in recent years was a rapidly rising trend. It made a great threat to human health, social and family to bring a heavy economic burden. China’s disease surveillance system data show that CHD mortality since the1990s has been an upward trend. In1991the mortality rate is25.3/100000. In1998has risen to42.7/100000. An increase of about50%in seven years, the world myocardialinfarction mortality has become the second ranks after cancer. So, effective prevention and treatment of CHD has become an important research topic in today’s worldwide. In our study, the first part we research Guanxinping adjustment for AS golden hamster from an overall perspective, in the second part we research Guanxinping as a role of protecting vascular endothelial cells and anti-apoptosis from the molecular perspective.Part1The effect of Guanxinping on AS golden hamsterHigh fat diet feed to the golden hamster. Detecting through regular blood parameters and pathological indicators, we established atherosclerosis golden hamster model. Continuous given Guanxinping six weeks, then took the blood test and chose liver, aortic pathological examination.The first section, we selected the the general indicators to evaluate and analyze the model, for inspecting Guanxinping role in terms of weight, blood lipids, aortic pathology, and liver histopathology. Results showed that the Guanxinping reduced the weight of AS golden hamster. At the same time,Guanxinping adjusted TG, TC, LDL levels. Liver and aorta damage caused by high fat diet also could be significant repaired.The second section, our reserch focus on hormone levels of golden hamster in blood. The results showed that Guanxinping reduced oxidative stress levels in AS golden hamster of MDA levels and increased SOD levels. It also could adjust procoagulant and fibrinolytic system in AS golden hamsters, increase t-PA levels in peripheral blood and reduce the level of PAI. Guanxinping reduced the peripheral blood of TXB2levels, and increased the levels of6-Keto-PGF1α. In addition, the drug reduced the ratio of the TXB2/6-Keto-PGF1α. Guanxinping also reduced the TNF-α levels in the AS golden hamster in peripheral blood, increased the level of NO, as playing a role of anti-inflammatory effect.The third section. By immunohistochemical methods, we examined the ICAM-1, TNF-α, NF-κB and IL-10of AS golden hamster in artery and investigated the aortic apoptosis. The results showed that, After the administration of drugs, ICAM-1, TNF-α, NF-κB expression levels can be reduced, but IL-10expression without significant changes. At the same time, Guanxinping could anti-apoptosis of the AS golden hamster aorta, with the effect to protect the aortic structure.Part2The mechanism of Guanxinping to protect the vascular endothelial cellsIn this part. We cultured human umbilical vein endothelial cells, and used serum pharmacology to prepare rabbit serum containing Guanxinping. We cultured vascular endothelial cells with serum containing Guanxinping. The H2O2was used to induce cell injury. We used biochemical technology to inspect the effect of Guanxinping on vascular endothelial cells which constitute an important part of vascular intimal under oxidative damage.The first section, we filter the damage concentration of H2O2. MTT assay with different concentrations of H2O2injury of HUVEC in order to investigate the appropriate concentration of H2O2. The results showed that different concentrations of H2O2cultured with HUVEC, we found that the concentration of H2O2during50-100umol/L is appropriate.The second section. Cultured with serum containing Guanxinping, then vascular endothelial cells injuryed by H2O2. We observed the cell culture medium for oxidative stress changes. The results showed that after culturing HUVEC with serum containing Guanxinping, and then cultured with H2O2for24h, Guanxinping could regulate oxidative stress, reduce cell culture medium MDA levels and increase SOD levels. In addition, It could also reduce the levels of ET and increase NOS levels in the culture medium.The third section. Cultured with serum containing Guanxinping, then vascular endothelial cells injuryed by H2O2. We observed the cell culture medium for inflammatory cytokine level changes. The results show Guanxinping could regulate the secretion of inflammatory factors, reduced IL-1of IL-6and TNF-α levels in the cell culture medium, as a important role for reducing the levels of inflammatory factors.The fourth section. Cultured with serum containing Guanxinping, then vascular endothelial cells injuryed by H2O2. We observed HUVEC apoptosis induced by H2O2detected by flow cytometry and Hoechst33258. The flow cytometry results showed that Guanxinping could reduce the rate of apoptosis of HUVEC. Hoechst33258fluorescent staining results showed that Guanxinping improved significantly the nuclear chromatin condensation status during apoptosis.The fifth section. After We detected Bax、 Bcl-2and Caspase-3levels in HUVEC by immunohistochemistry. Results showed that Guanxinping could reduce cell apoptosis, Bax and Caspase-3protein expression and increase Bcl-2expression. The effect of Guanxinping resistance to apoptosis induced by H2O2might be important in the process of regulating apoptotic proteins pathway to achieve, such as Bax, Bcl-2and Caspase-3.