| ObjectiveThe purpose of this experiment is to explore a role of low-dose heparin on early intervention in septic rats . It can be demonstrated by detecting the platelet, APTT, the inflammatory mediators TNF-αlevels ,observing the pathological changes of lung tissue,making pathology score,and observing the mortality rate in 24 hours through a subcutaneous injection of low-dose heparin at different time points in rats. So we can pave a successful road to the treatment of clinical sepsis disease in using heparin from an aspect of theoretical and experimental basis.MethodHealthy adult 72 SPF Wistar rats were randomly divided into three groups, namely sham operation group (sham), CLP group (CLP) and the heparin group (heparin), n = 24. After an anesthesia laparotomy, sham operation group only was changed the cecum, CLP group was underwent cecal ligation and puncture, heparin group was received a subcutaneous injection of heparin (14U/kg/12h) immediately after CLP. Each group after another by 6h, 12h, 24h three time points were divided into three subgroups, each subgroup 8, they were tested blood platelets and APTT in blood routine test, the determination of plasma TNF-αcontent by using enzyme-linked immunosorbent assay (ELISA) method, samples of lung tissues by HE staining to observe pathological changes in lung tissue and make pathology score.The other 40 rats in the same conditions were divided into CLP group and the heparin group, n = 20, given the same treatment factors, and observed for 24-hour mortality rate of rats in sepsis. Results1) Platelet count: CLP group and sham operation group, platelets decreased significantly in the 12h and 24h (P <0.01), heparin group at 24h also reduced (P <0.01), but more than the CLP group (P <0.01).2) APTT: 24h, CLP group than in the sham group was significantly longer (P <0.01), heparin group than in the sham group also extended (P <0.01), but shorter than the CLP group (P <0.01).3) The plasma TNF-αcontent: CLP group and heparin group were higher than sham operation group at all time points (P <0.01), but in the 12h and 24h heparin group were lower than the CLP group (P <0.01).4) Lung pathology: In optical microscope, sham group kept normal structure of lung tissue with no congestion and inflammatory cell infiltration; CLP group had a observation of significantly widened alveolar septum, obvious bleeding and inflammatory cell infiltration in lung; heparin group's lung congestion and inflammatory cell infiltration significantly reduced compared with CLP group. Compared with the sham group, lung pathology scores of CLP and heparin group were increased in the three time points(P<0.01), at 24h, CLP group's score was the highest one (P<0.01),and heparin group's pathology score was lower than CLP group(P<0.01).5) 24-hour mortality: In CLP group, seven rats died and 13 survived within 24 hours, the mortality was 35%, while in heparin group only one rat died and 19 survived within 24 hours, the mortality was 5%, This test results showed that the heparin group's 24-hour mortality was significantly lower than the mortality in CLP group (χ2= 5.625, P = 0.018) with statistical significance.ConclusionIt can be concluded from this experiment that the low-dose heparin can rebound platelet count, shorten APTT values in rats at 24h after CLP, and reduce blood levels of TNF-αin the last two time points 12h and 24h, and can also reduce lung pathology at 24h. From these, we can say that the low-dose heparin regulate the network of coagulation and inflammation system in sepsis, reduce the occurrence of coagulation disorder and thereby reduce inflammation in lung tissue's damage, so early intervention of heparin play a role in preventing sepsis from developing in a certain extent, and can reduce mortality of septic rats in 24 hours which will benefit the prognosis of sepsis in a positive direction. |
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