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Objective To study the different periods of LMWH administration intervention ANP rat serum PAF, ET-1, NO, TXA2, PGI2 change impacts observed ANP rats with 72-hour survival time and statistical survival, explore different times of the day LMWH administered intervention on ANP large the role of the rat pancreatic microcirculation, LMWH best time for the clinical treatment of severe acute pancreatitis (severe acute pancreatitis, SAP) to provide a theoretical and experimental basis. 160 Methods SD rats, weighing 250-300g were randomly divided into the LMWH group (L), n = 64; saline (normal saline, NS) group (N), n = 64; sham-operated (sham operation, SO) group (S group), n = 32. L and N group was randomly according to the different administration point in time and then each divided into four groups L0H group (n = 16), L6H group (n = 16), of L12h group (n = 16), L18h group (n = 16 ) and N0H group (n = 16), N6H group (n = 16), N12h group (n = 16), N18h group (n = 16). L and N rats were 5% sodium taurocholate (0.1 ml/100 g) micro-pump ANP model was established by injecting retrograde cholangiopancreatography the S rats open only changes the pancreas modeling. L rats the ANP model 0:00 (L0H), 6 (L6H), 12 (L12h), 18 (L18h) subcutaneous injection of LMWH (You Nishu 50IU/ml, 10IU/100g weight / ), repeated every 6 hours injection until 24 hours after modeling; rat ANP group N into a mold at each time point after subcutaneous injection of NS, the same group L (0.2 ml / 100 g body weight / time). Three groups of rats at 24 hours after the ANP model, randomly selected for each group of rats 1/2 of the number of open again to observe the amount of ascites and color, general change in intra-abdominal the saponification spots and pancreas, take the portal blood to detect blood PAF, TXB2 (TXA2) ,6-Keto-PGF1α (PGI2), ET-1, NO, blood AMY blood Ca2 concentration, pancreas HE staining pathological examination parallel pancreatic tissue pathology score, transmission electron microscopy of pancreatic microvascular thrombosis, group the remaining 1/2 the number of rats continue to fast, free drinking water, observed 72 survival time and survival rate statistics. Intraperitoneal general situation: L, N, S, three groups of rats ANP modeling 24h random from each of 1/2 again laparotomy findings: L rats intraperitoneal visible yellow slightly turbid ascites, mesenteric dot saponification spots , pancreas point sheet hemorrhage and necrosis; the N rats intraperitoneal visible light red bloody ascites, formation of mesenteric poorly saponification spots, pancreatic haemorrhage, necrosis; S rats did not show these changes. 2 blood parameters of serum Ca2 and AMY: L rats at various time intervals blood Ca2 concentration higher than the of N rats corresponding period Ca2 concentration (P lt; 0.01); the L rats at various time intervals blood AMY concentration always make less than N group, each period AMY concentration (P lt; 0.01); L and N two groups of rats blood Ca2 the AMY concentration group within the different times of the day or two was no significant difference (P gt; 0.05) Blood PAF: L rats at various time intervals set of blood concentration of PAF were lower than the corresponding period of N rats group concentration (P lt; 0.01), L0H group (6.559 ± 0.748) and, L6H group (6.369 ± 0.712) were lower than L12h L12h group (7.323 ± 0.323) (P lt; 0.01) and L18h group (7.343 ± 0.615) (P lt; 0.05); L0h group (6.559 ± 0.748) and of L6H group (6.369 ± 0.712) (7.323 ± 0.323) and the of L18h group (7.343 ± 0.615), no significant difference (P gt; 0.05) Blood TXB2 (TXA2), 6-keto-PGF1: L rats blood TXB2 (TXA2), 6-Keto-PGF1 group concentration of each period were lower than the corresponding N rats at various time intervals group of blood TXB2 (TXA2), 6 - keto-PGF1 concentration (P lt; 0.01); L0H blood TXB2 (TXA2) (39.386 ± 2.089) and blood 6-keto-PGF1 (52.550 ± 1.047) and the L6H group blood TXB (2TXA2 () 40.220 ± 2.338) and blood 6-keto-PGF1 concentration (52.374 ± 1.604) were lower than the L12h blood TXB2 (TXA2) (57.384 ± 3.838) (P lt; 0.01) and blood 6-Keto-PGF1 (57.743 ± 1.504) (P lt; 0.01) L18h blood TXB2 (TXA2) (57.924 ± 4.262) (P lt; 0.01) and blood 6-keto-PGF1 concentration (58.093 ± 1.189) (P lt; 0.01); the L6H L0H between L12h group L18h group between the group of blood TXB2, blood 6-keto-PGF1 concentration showed no significant difference (P gt; 0.05) Blood ET-1, NO: L rats blood ET-1 concentration of NO each time group were lower than N rats each of the corresponding periods set of blood ET-1, NO concentration (P lt; 0.05); L0H group (81.753 ± 7.227) and L6H group of rat serum ET-1 concentration (80.358 ± 8.270) were lower than the L12h group (92.971 ± 6.849) (P lt; 0.05) and L18h group (95.553 ± 6.903) (P lt; 0.01); but between the L0H group and L6H group the and the L12h group L18h group between blood group ET-1 concentration was no statistical difference (P gt; 0.05); each period rats NO concentration pairwise comparisons were not statistically different (P gt; 0.05). 3 Histological examination of pancreatic tissue pathological examination of HE staining: N rats visible rupture of pancreatic acinar cells, acinar structure disappeared, large patchy hemorrhage and necrosis, a large number of neutrophil infiltration; L rats visible pancreatic acinar fine rupture less than the N group into points haemorrhage necrosis, the surviving less normal acinar cells, more neutrophil infiltration; S rats pancreas gland structure, the integrity of the acinar cells, no necrosis , bleeding and inflammatory cell infiltration. Pancreatic tissue pathology Rated: L0H out group (6.000 ± 0.926), L6H group (6.500 ± 1.069), of L12h group (8.750 ± 1.283), of L18h group (8.250 ± 1.670) in rat pancreatic histopathological scores lower than that of the N corresponding each time N0H group (10.000 ± 0.756) (P lt; 0.01), N6H group (10.250 ± 1.282) (P lt; 0.01), N12h group (10.500 ± 1.195) (P lt; 0.01)., N18h (8.250 ± 1.670) (P lt; 0.01) rats pancreatic histopathological scores; L0H group (6.000 ± 0.926) and of L6H group (6.500 ± 1.069) in rat pancreatic histopathological scores were lower than the of L12h group (8.750 ± 1.283) (P lt; 0.01) L18h group (8.250 ± 1.670) (P lt; 0.01), but L0H group (6.000 ± 0.926) and of L6H group (6.500 ± 1.069) and L12h group (8.750 ± 1.283) and of L18h group (8.250 ± 1.670) between the rat pancreatic tissue pathology score was no significant difference (P gt; 0.05). TEM: N rats visible pancreatic acinar cell structural damage, normal pancreatic acinar disappeared, a lot of necrotic tissue, a large number of micro-thrombosis, blood vessels rupture, the remnants of microvascular neutrophil retention; L rats visible intravascular micro-thrombosis less than the N group, the remaining small amount of normal pancreatic acinar cells. S rats intravascular thrombosis, pancreatic acinar cells, structural integrity. Three groups of rats at various time intervals pancreatic histopathological scores and blood PAF ET-1/NO TXB2/6-Keto-PGF1α (TXA2/PGI2) correlation coefficient r 0.808,0.835,0.736 showed a positive correlation (P lt; 0.01). Survival time in rats: L group 48h, 72h survival rates were 71.87% and 46.87%, respectively, higher than the N group 48h (43.75%) (P lt; 0.05) and 72 h (21.87%) (P lt; 0.05) survival rate, SO group, but less than 48 h (100%) (P lt; 0.05) and 72 h (100%) (P lt; 0.05) survival. Conclusion (1) LMWH subcutaneous injection by reducing the ANP rat blood PGI2, ET-1, NO, TXA2, concentration and ET-1/NO TXA2/PGI2 improve pancreatic microcirculation, thereby reducing pancreatic pathological damage. (2) reduce the ANP rats blood of ET-1, the ratio of the NO, TXA2, PGI2 concentration and ET-1/NO, TXA2/PGI2 reduce pancreatic pathological damage, LMWH 0:00 rat ANP into a mold and 6:00 better than subcutaneous administration until the beginning of 12:00 and 18:00 before subcutaneous administration. (3) LMWH can effectively improve the ANP rat prognosis, improve survival.
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