Panax Notoginseng Saponins Endotoxic Shock Rats Myocardial Protection Mechanism

Septic shock is a familiar emergency and severe case,with a high mortality about 50%-60%. The common cause leading to which is the infection of Gram-negative bactFlea and endotoxin plays an important role in the occurrence and the maintenance of the shock. When the shock occurs, The bactFlea can be killed by antibiotic ,but sometimes the shock can not get to retrieve ,even aggravating. Because a large quantity of endotoxin ,e.g. LPS,was released from bactFlea which is dead or killed by the medicine.The complex of LPS and LBP,is presented to the receptor of D14 on the surface of cell by glycon-monophosphoinositide. CD14 is short of transmembran and intra-cellular structial domain,but can transmit signal to the intra-cellular on the help of transmembrane signaling protein.TLR is key transmembrane signaling protein,while TLR4 is homologous modif of human TLR,which exist in monocyte , neutrophil cardiocyte vascular endothelial cell mechanocyte as a key receptor participating in natural immunity .The complex of LPS-CD14 and TLR4 with the help of MD2,transmits signalto intra-cellular.MyD88 is activated by the combination of TLR4 intra-cellular region and MyD88-protein,then activatingIRAK TAK1 and I-kB , increasing synthesis and deliverance of TNF-a ,inducing other inflammatory factor.All of which results in cellular tissue damage, microcirculation disturbance, then to cause multiple organs,such as heart, lung, liver, kidney, intestinal tract etc. dysfunction syndrome.Among the total the heart functional lesion is an important caused by shock. For the reason ,it has been a hot spot to research how to degrade the mortality of septic shock.Traditional Chinese Medicine consider that Endotoxin shock resembles syncope syndrome.It is caused by invasion of external pathogenic factors, deeply attacking yingfen and xuefen, then to consume qi and xue and blending pathogenic heat,toxin substance and blood stasis.It leads to exhaust the funtion of zang-organs and fu-organs, separate yin and yang, loss of yang-qi. And the main therapeutic methods were to remove toxic substance and active blood stasis.Radix notoginseng has the effect of eliminating blood stasis,stopping bleeding , and removing toxic substance and relieving edema. Modern research indicates the main ingredient of Radix notoginseng is total Saponins of Radix Notoginseng,besides the Radixsaponin Rb1 with the effect of diminishing inflammation and relieving pain,the Radixsaponin Rg1 and Notoginsenoside R1 with the effect of anti-platelet aggregation and anti-thrombus. The animal expFlements suggest that total Saponins of Radix Notoginseng has the function in protecting on the cardiocyte of Endotoxin Shock Rats,so it is with great important clinic sense to deeply study the protective mechanism .This research is composed of two sections as animal experiment and cell culture experiment.1.Animal experiment1.1 Objective:To observe the curative effect of total Saponins of Radix Notoginseng in endotoxin shock,and to approach the mechanism initially of it's function of anti-endotoxin shock.1.2 Method: SD rats were divided into six groups in random:NS group, endotoxin group, total Saponins of Radix Notoginseng group of equivalent dose, total Saponins of Radix Notoginseng group of large dose, fleabane group, Taurine group.And the endotoxin shock model was copied after it was treated with correlated drugs for three days. Then the index of MBP, HR, LVSP were observed ; the cTnI in blood plasma, the TNF-a of serum and myocardium wered detected, and the expression of TLR4 at mRNA level in the myocardium by PT-PCR method wered observed.1.3 Result:Total Saponins of Radix Notoginseng 1)can improve the LVSP of endotoxin shock rats obviously, take an important role in stabilizing blood pressure and heart rate, and can delay the occurrence of shock. 2)can degrade the level of plasma cTnI in endotoxin shock rats obviously,and relieve the inflammatory reaction of myocardium.3)can restraint the expression of TLR4mRNA in myocardium,thereby restraint formation of TNF-αin blood plasma and myocardium.1.4 Conclusion:Total Saponins of Radix Notoginseng restraints the expression of TLR4mRNA in myocardium, thereby restraint formation of TNF-αin blood plasma and myocardium,lessen cellular damage of myocardium, thus stabilize blood pressure , heart rate and improve heart function.2. Cell culture experiment2.1 Objective:To observe the influence between Total Saponins of Radix Notoginseng in blood serum and cytoactive of cardiac muscle, and to approach the potential mechanism initially.2.2 Method: the myocardial cell of neonate rat wered divided into six groups,and rat's blood serum wered added into, the control group with 10% NS,and the rest with 10% NS ,total Saponins of Radix Notoginseng of equivalent dose, total Saponins of Radix Notoginseng group of large dose, fleabane, Taurine.Then suscitated them with endotoxin .To observe the effect on the increment of myocardial cell by MTT reduction assay and the effect on the expression of TLR4,TNF-αat protein level with immunohistochemical methods and image analysis methods.2.3 Result:The animal blood serum with Total Saponins of Radix Notoginseng 1)can obviously improve the restraint on the increment of myocardial cell from endotoxin .2)can restrain the expression of TLR4 and TNF-αprotein in myocardial cell,which was stimulated by endotoxin.2.4 Conclusion:Total Saponins of Radix Notoginseng can obviously degrade the expression of TLR4 and TNF-αprotein of myocardial cell,thus restrain the occurrence of inflammatory mediators to achieve the function in protecting myocardial cell.