The Study On Mechanisms Of Septic Spleen Apoptosis And The Treated Effects Of Salvia Miltiorrhiza On Rat

Objective This study was performed cecal ligation and puncture(CLP) on rat to induce sepsis and odserved consecutively the effect of splenic tissue; To investigate the effects of the dynamic level change of TNF-a and OFR in spleen apoptosis, To discusses the mechanism of spleen apoptosis ;To observe the treated effects of salvia miltiorriza in spleen apoptosis.Provides certain theory basis for clinical treatment of sepsis.Methods 96 male rats weighing 180-250g were utilized and assigned randomly to 4 groups:control group(Control),sham operated group(Sham),cecal ligation and puncture group (CLP),and salvia miltiorrhiza treated group(Salvia).Each grouop divided 4 time point,and each time point apportioned 6 rats randomly.Rats in Control group were only anesthetized by injected 3 percent pentobarbital sodium into abdominal cavity with dosage 30mg/kg.To rats in Sham group,the cecum was located but neither ligated nor punctured. To rats in CLP group,the cecum was ligated and punctured with a 20-gauge needle in three places to induce sepsis.Rats in Salvia group received salvia militiorrhiza through abdominal cavity injection when CLP was nearly finished.Rats were anesthetized and killed at every time point to obtain samples.Blood was obtained though abdominal aorta puncture,and the spleen was resected and divided two equal parts of which one was to make hepatic tissue homogenate and another was for pathological investigation.MDA,GSH-PX and TNF-a was tested according to methods of thibabituric acid, enzymology and ABC-ELISA,Respectively. The technique of terminal deoxynucleotidyl transferase-mediated dUTP nick end labling(TUNEL) was used to detected spleen lymphocyte apoptosis.The impacting on spleen lymphocyte apoptosis were filtrate by multinomial linear regressions.Result 1.affirmation of the succeeded CLP model: severity of intraabdominal inflammation was correlated with time after CLP and reached peak at 48h with much more purulent or bloody exudation in the cavity. Comparing with CLP group, no obviously peritonitis was found in Control and Sham group2.The changes of serum and splenic TNF-a levels: TNF-a levels both in CLP and Salvia group began to increase at 6h , reached their peak at 24h and 48h, respectively. However, both serum and tissue TNF-a levels in Salvia group was significant lower than those in CLP at 24h (p < 0.01).3.The changes of serum and splenic MDA levels: MDA levels in CLP and Salvia group began to increase at 6h and 12h, respectively.and reached their peak at 48h, However, serum MDA levels in Salvia group was significant lower than those in CLP atl2h and 24h, respectively, (p < 0.01). splenic tissue MDA levels in Salvia group was significant lower than those in CLP at 6h,12h and 24h, respectively (p < 0.01).4.The changes of serum and splenic GSH-Px levels: GSH-Px levels in CLP and Salvia group began to decrease at 6h and 12h, respectively,and reached their minimum at 48h. However, both serum and tissue GSH-Px levels in Salvia group was significant higher than those in CLP at 6h,12h and24h, respectively, (p < 0.01).5.The changes of splenic lymphocyte apoptosis: The splenic TUNEL-positive cell rates in CLP group increased at 6h and reached its peak at 24h. The splenic TUNEL-positive cell rate in Salvia group was significant lower than those in CLP group at 12h and 24h, respectively, (p < 0.01).6.The result of filtrate by multinomial linear regressions: The factor influenced spleen lymphocyte apoptosis were levels of MDA in splenic tissue and serum,TNF-a in splenic tissue and splenic tissue GSH-Px.Conclusion 1.The rat model by sepsis. can be established successfully by the method CLP for experimental observation .2. The spleen is one fo the target organs in MODS by sepsis, spleen lymphocyte apoptosis is main change of injury encountered in sepsis , and sepsis causes the TNF-a level increased and the oxygen free radical metabolism disorder has in the mechanism in it to play the vital role.3. The salvia miltiorrhiza may protect spleen organizes through decreasing the TNF-a level of organism, adjustment free radical metabolism, prevents immunosuppression which the spleen cell is excessively apoptosis causes.