Effects Of Shen-Fu Injection On Myocardial Injury In Rats With Septic Shock

Objective: To investigate the protective effects and the possible mechanism of Shenfu injection(SF) on myocardial injury in rats with septic shock and to observe the expression of nuclear factor-κB (NF-κB), tumour necrosis factor-α (TNF-α) and interleukin-10(IL-10).Methods: 48 wistar rats were randomly divided into 3 groups with 16 animals in each group: sham-operation group(C group) only underwent laparotomy; septic shock group(S group) underwent cecal ligation and punctures(CLP); and Shenfu treated group(SF group) received Shenfu injection 8ml·kg^-1·h^-1 iv for 2 hours followed by CLP . Each group was further divided into 2 subgroups: 6h and 18h subgroups base the time when blood sample was taken and the animals was sacrificed. Using CLP method duplicated septic shock model: Under aseptic conditions, a 2-cm midline laparotomy was performed to allow exposure of the cecum with adjoining intestine. The cecum was tightly ligated with a 1.0 silk suture at its base, below the ileocecal valve, and was perforated twice with an 18-gauge needle (top and bottom), after anaesthesia. Right femoral artery and vein were cannulated for MAP monitoring and fluid infusion. Myocardium of rat left ventricle were drawn to observe pathology under optical microscope and to observe the change of ultrastructural organization under electron microscope. Immunohistochemical staining method of SP was used to locate the expression and distribution of NF-κB in myocardium. The plasm levers of TNF-α and IL-10 were determined by Enzyme-linked immunoadsordent assay (ELISA). The contents of maleic dialedhyde (MDA) was measured by thibarbituric acid method (TAB) in myocardium homogenate.Results: Compared with C group, MAP gradually decreased and Heart Rate gradually increased at 6h after CLP in S group (P<0.01) ; at 18h, both MAP and Heart Rate markedly decreased (P<0.01) . However, MAP started decreasing at 18h in SF group (P<0.05 ). The average OD value of NF-kB protein was significantly higher in IR group than that in SF group (0.1897±0.0214 vs. 0.1214±0.0118, P<0.01) and in C group(0.1897±0.0214 vs. 0.1029±0.0109, P<0.01). The NF-κB protein was mostly located at cellular nuclei and sparsely at cellular plasm. The contents of maleic dialedhyde (MDA) was significantly higher in S group than those in SF group (P<0.01)and in C group(P<0.01) . The levels of TNF-a in the plasma were obviously higher in S group than those in SF group^O.Ol) at the same stage. The difference of the levels of IL-10 in the plasma was not statistically signigicant between S group and SF group (P>0.05) at 6h, but the levels of IL-10 were obviously higher in S group than those in SF group(P<0.01) at 18h. There were less myocardial injury in SF group than that in S group under optical microscope and electron microscope at the same stage.Conclusion: Shenfu injection could protect myocardium from the injury induced by septic shock through inhibiting the activity of NF-kB and the expression of TNF-a and IL-10 to prevent excessive inflammatory reaction and immunity suppression . It also relieve the degree of lipid peroxidation of myocardium to inhibit the levels of MDA and lessen mitochondria injury of myocardial cell.