Experimental Study Of Low Molecular Weight Heparin And Lithospermum Erythrorhizon Combination On Expression Of Rats With Severe Acute

IntroductionSevere acute pancreatitis (SAP) had a character of quick onset, serious symptoms, rapid course development and conventional medical treatments show low efficacy in the treatment of SAP. In the present treated circumstances, its mortality reached 22.7% still.So finding ideal therapeutic drug has become the key point of this disease treatment. In the recent years, the application of lower molecular weight heparin (LMWH) on the treatment of experimental and clinical aspects of SAP have had a series of profound study by scholars at home and abroad. They found that LMWH can improve microcirculation and refrain inflammatory reaction of severe acute pancreatitis in rats.It has recognized that Lithospermum erythrorhizon has many activity, such as anti-inflammatory, regulating immune, anti-tumour and protecting liver. It has not reported that whether it is valid that Lithospermum erythrorhizon to SAP. In this study, we researched low molecular weight heparin and Lithospermum erythrorhizon combination on expression of serum SOD,MDA,NO and renal tissue's HSP70 of Rats with Severe Acute Pancreatitis, and investigated if there was synergetic effect of combined treatment on SAP.ObjectiveTo investigate the effect of low molecular weight heparin and Lithospermum erythrorhizon combination on the mortality of Rats with Severe Acute Pancreatitis, dynamic changes of serum SOD,MDA,NO at different time points, expression of renal tissue's HSP70. Further discusses the mechanisms of low molecular weight heparin and Lithospermum erythrorhizon combination on SAP and effective measures of relieving SAP in order to provide theoretical basis for clinical treatment.MethodsA total of 100 Wistar rats, weight 280-350g, aged 10-12weeks, half males and females, were randomized into 5 groups, each 18 in every group:the sham operated control group(group A only laparotomy),SAP group(group B model group),treatment with low molecular weight heparin group(group C heparin group),treatment with Lithospermum erythrorhizon group(group D Lithospermum erythrorhizon group),treatment with low molecular weight heparin and Lithospermum erythrorhizon combination group(group E compound group). Severe acute pancreatitis model was inducted by biliopancreatic retrograde injection of sodum taurocholate solution. After SAP model was manufactured successfully, then group C were treated immediately with LMWH(100iU/kg/12h), group D were lavaged immediately with Lithospermum erythrorhizon(3g/kg),2 times each day, group E were treated with LMWH and Lithospermum erythrorhizon combination and the control group were treated with equivalent normal saline. Every group anesthetized six rats at different time points(12h,24h,48h and 72h) after operation. The monocular enucleation was performed to get blood and detected serum SOD,MDA,NO and detected expression of renal tissue's HSP70 after operation. Meanwhile compared the mortality of five groups at 24h and 3d after operation. Data are expressed as mean±standard deviation (SD) for continuous variables.The measurement data were compared with F-test and the mortality were analyzed byχ2-test. P<0.05 was considered statistically significant.Results:(1)Observation of the mortality:Sham operated control group all survived after operation. The mortality of 24h:group B was 55.56%, group C was 11.11%,group D was 27.78% and group E was 11.11%;The mortality of 3d:group B was 88.89%,group C was 16.67%,group D was 50.00% and group E was 11.11%.group C, group D and group E showed significant difference compared with group B(P<0.05,P<0.01).This indicated that SAP rat model has established.(2)SOD activity:There was significant difference in SOD viability of group compared B with group A (P<0.01); The SOD activity of group C and group E showed obviously difference compared with group B (P<0.01)at different time points after operations. There was significant difference between group D (48h 72h) and group B (P<0.05, P<0.01).(3)MDA level:There was no significant difference between every group(12h) and group B (P>0.05);At 24h,48h and 72h time point,group C and group E showed obviously difference compared with group B (P<0.01);There was significant difference between group D (48h 72h) and group B (P<0.05). (4)NO content:There was no significant difference between every group(12h) and group B (P>0.05);At 24h time point,group A showed obviously difference compared with group B (P <0.01);There was no significant difference among other three groups (P>0.05), There was great difference between every group(48h 72h) and group B(P<0.01, P<0.05)(5) Renal tissue's HSP70:The expression of HSP70 in each group at 24h was more than 72h. Statistics indicates a distinctive difference (P<0.05,P<0.01); the expression of HSP70 was significantly increased in group B compared with group A(P<0.01),The positive expression areas of HSP70 in group C, group D and group E decreased significantly compared with group B, and there was significant difference(P<0.01).ConclusionLow molecular weight heparin and Lithospermum erythrorhizon combination can effectively decrease the mortality of SAP rat, increase the activity of SOD, suppress the production of MDA, increase the level of serum NO, decrease the expression of HSP70 in renal tissue,obviously improve pancreatic microcirculation and restrain inflammation. There have therapeutic effects to SAP. The mechanism may be related to dilate vessels, anti-free-radical injury and protect the stable structure of cell membrane.These results indicated that single therapy with Low molecular weight heparin or Lithospermum erythrorhizon had certain curative effect, but the effect of Lithospermum erythrorhizon was inferior to low molecular weight heparin.Low molecular weight heparin and Lithospermum erythrorhizon combination have effective treatment to SAP, but the combination have no evident synergistic effect.