Effects Of Bushen Fumai Liquor On Synthesizing And Releasing Nitric Oxide And Endothelin In Hypoxic Endothelial Cells

Effects of Bushen fumai liquor on synthesizing and releasing NitricOxide and EndotheIin in hypoxic Endothelial CellsAbstractObjective: To observe and study the effects of Bushen fuIni liquor on thesynthesizing and releasing of Nitric Oxide nyO) and Endothelin (ET), the acivityof Nitric Oxide Synthase NOS), the exPression of eNOSmRNA and ET-lmRNA. in cultUred Human umbilical Vein Endothelial Cells (HUVECs) induced byhyPoxia.Method: The cultured HUVECs were randomly divided into four groups: theblank (nonnoxic) grouP, the model (hyPoxic) grouP, the westem medicine grouP(hyPoxiWtC) and chinese medicine grouP (hyPoxia+Bushen fumai liquor).Respectively, the l0% blank rabbit serum, l0% blank rabbit serum, l0% rabbitserum contaning weStCrn drugs and 10% rabbit serum containing chinese drugswer administrated to the abOve four grouPs of HUVEC. Except the b1ank grouP,the other grouPs were exposed tb hrpoxia (95%N2+5%CO2) for four hours. Firstly,the product of NO. ET and the activity of NOS from cultured HUVECs of theabove four grouPs were respectively determined. Then the tOtal RNA wasextracted frOm the cultated HUVECs with the method of guanidinium thiocynate.Fiedly, the reverse transcriPtion and polymerase chain reaction (RT-PCR) wereperformanced on every grOuP. RTPCR products were fractionaed into sizethrOugh a l.0% agarose gel, electrophoresis bands with ethidium bromide werevisualized under the ultraViolet op) ligh and photograPhed. Bands on film werescaned by the densitometef, and then were quanified on comPuter by the imageanalysis sobo.Results(l)NO and NOS: Both NO level and NOS activity of the hyPoxic grouP decreasedl 83more significanly than those of the blank grouP (p<0.0l),and thOse of the chineseand westem medicine grouPs increased more significanly than those of thewestem medicine grOuP (P<0.0l,p<0.05),but there was no significgh differencebe'tWen the chinese and westem medicine grOuPs (P>0.05).(2)ET: The level of ET in hyoPxic grouP increased more significanly than that ofthe blank grouP (P<0.0l), but that of the weStern medicine grouP had no moresignifican differences than tha of the hyPoxic grouP (P>0.05), the chinesemedicine gfOuP decreased more significantiy than that of the hyPoxic grouP(P<0.001), and the chinese medicine grouP was saperior to the westem medicinegrouP (P<0.01).(3)eNOSmRNA: The level of eNOS/GAPDHmRNA in the hyPoxic grouPdecreased more significantly than that of the blank grouP (P<0.0l ), that of both thechinese and westem medicine grouPs increased more significanly than that of thehypoxic grouP (p<0.0l,P<0.05), and the chinese medicine grouP was suPerior tothe westem medicine grouP (P<0.05).(4) ET-lWA: The level of ET-l/GAPDHmRNA in the hypoxic grouP increasedmore significanly than tha of the blank grouP (p<0.0l), tha of the chinese andwestem medicine grouPs decreased more significan1y than that of the hypoxicgrouP (P<0.0l,p<0.05). The chinese medicine group was superior to the westemmedicine grouP (P<0.05).Conclu8ion The chinese medicine "Bushen Fumal liquor" could inhibit theUnbalance betWeen NO and ET synthesized and released by the hypoxicendothelial cells through decreasing ET or increasing NO, as showed that BushenFumai liquor had a significanly protective effect on the hyPoxic endothelial cells.