The Effect Of Calycosin For ET-1/NO On The Damage Of TNF-α In Human Umbilical Vein Endothelial Cells

Objective:Chronic kidney disease (chronic kidney disease, CKD) is a group of progressive development of chronic non-infectious diseases. CKD is a high prevalence of the common diseases worldwide. Because of its high prevalence, poor prognosis and a huge health care costs has become a major hazard to human health diseases. CKD patients may eventually develop chronic renal insufficiency, uremia, eventually requiring dialysis or kidney transplantation. Costly dialysis treatment, thus, delaying the progress of CKD, delay the progress of CKD patients to uremic blood purification treatment for the time, with important social, economic significance. Endothelial dysfunction exists in patients with CKD and CKD occurred throughout the development of the whole process is the key link in the pathogenesis of CKD, but also the basic characteristics of CKD patients. Moreover, endothelial cell dysfunction associated with atherosclerosis (AS) in various stages of CKD patients with cardiovascular events and mortality are closely related. Endothelial dysfunction is the pathological change of CKD, while the endothelial cell dysfunction is closely related with the development of CKD can also be manifested as hypertension, coronary artery disease, chronic heart failure, peripheral vascular disease, diabetes, etc. [2], they collaboration to accelerate the progress of deterioration of CKD. Therefore, protection of endothelial cells in the course of CKD is a delay, prevent chronic renal failure, the key to improving the AS has an important positive significance. Calycosin is a flavonoid drug, is an important component of Astragalus. According to reports, astragalus can scavenge oxygen free radicals, antioxidant, immune and so on. This article explores calycosin Astragalus monomer TNF-a induced on human umbilical vein endothelial cells (ECV-304) of nitric oxide (NO), endothelin-1 (ET-1) effects.Methods: DMEM culture medium at 37℃.5% CO2 conditions on the ECV304 cell line was amplified to a sufficient number, the adjusted cell density of 1×105,and divided into 7 groups:A group:control group, without any inducer; B group:TNF-α(40ng/ml) induced group; C group:TNF-α(40ng/ml)+losartan (10-5mol/1); D group:TNF-α(40ng/ml)+calycosin (0.1mg/ml); E group:TNF-α(40ng/ml)+calycosin (1mg/ml); F group:TNF-α(40ng/ml)+ calycosin (10mg/ml); G group:TNF-α(40ng/ml)+calycosin (20mg/ml). Were incubated for 6 hours, supernatant was collected after 24 hours, the supernatant by ELISA detection of nitric oxide (NO), endothelin 1 (ET-1) levels.Results:①compared with the control group, TNF-a reduced the ECV-304 NO levels (P <0.001), increased ET-1 levels (P<0.001);②losartan increased ECV-304 NO levels (P<0.001), reduced the ET-1 levels (P<0.001);③calycosin (0.1 mg/ml-10mg/ml) ECV-304 cells can promote NO production (P<0.001), reduced ET-1 production (P<0.001);④losartan potassium and calycosin (20mg/ml) ECV-304 cells for ET-1 and NO levels not significantly different ((P>0.05);⑤calycosin concentration (at 0.1 mg/ml-20mg/ml inside) and ECV-304 ET-1 secretion was negatively correlated (r=-0.02, P<0.001); and ECV-304 NO secretion was positively correlated (r=0.0075, P<0.001).Conclusion:1.TNF-a inhibited the endothelial cells NO, promote secretion of ET-1; 2. Astragalus calycosin monomer can promote TNF-a induced endothelial cell NO secretion, inhibition of ET-1 secretion, and within a certain concentration (0.1 mg/ml-20mg/ml) in a concentration dependent manner.