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Objective To study the dynamic expression and distribution of high mobility group box 1 (HMGB-1)in diffuse axonal injury (DAI)in rats and to clarify its involvement in the inflammatory reaction after DAI in rats,in order to provide new targets for the clinical treatment of DAI.Methods A DAI model was established using a coronal rotation device and evaluated by HE,Glees-Marsland silver staining,and Mallory phosphotungstic acid hematoxylin staining.Immunohistochemistry,Western blot and RT-PCR were used to detect the expression and distribution of HMGB-1 in the cortex of DAI rats at 6 h,1 d,3 d and 7 d.And TUNEL was used to examine the apoptosis of neurons in DAI rats.Results Immunohistochemical results showed that at 6 h and 1 d after DAI,the number of HMGB-1-positive cells decreased,but at 3 and 7 d it began to increase.Western blot also showed that during the early stage after DAI (6 h and 1 d),the level of HMGB-1 protein in the cortex was significantly lower than that in the control group,but at the late stage (3 and 7 d)after DAI it significantly increased compared with that in the control group until 7 d.RT-PCR showed that at 6 h after DAI there was no significant increase in the level of HMGB-1mRNA,but at 1 d there was a slight increase compared with the control group;at 3 and 7 d,it showed an obvious significance.TUNEL staining indicated that the significant neuronal apoptosis appeared as early as 6 h after DAI,and reached the peak at 3 d;it started to decrease at 7 d but still remained at a relatively high level.Conclusion The dynamic expression and distribution of HMGB-1 showed significant changes with the time course after DAI in rats.They decreased at the early stage but increased at the late stage.At the early stage, HMGB-1 is mainly passively released by the necrotic neurons,and at the late stage it may be actively secreted by the active inflammatory cells.HMGB-1 may mediate the post-DAI neural cell apoptosis by inducing the inflammatory reaction.
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Objective To investigate effect of Dahuang Fuzi decoction on alveolaur epithelial barrier in rats with lung injury with severe acute pancreatitis. Method Ninty-six health SD rats were randomly divided into three groups: sham operation group, SAP-ALI group, Dahuang Fuzi decoction group, and then according to the time point of sacrifice after operation, each group was subdivided into 3,6,12,24 hour subsets ( each, n = 8). After the belly of a rat in the sham operation group was cut open, the pancreas was flipped several times,and then a stoma was made in the jejunum to form its fistula. In the SAP-ALl group,1 mL/kg sodium taurocholate was reversely injected into the pancreatobile duct to establish the model of SAP, and then the jejimum fistula was performed. The SAP-ALI model in Dahuang Fuzi decoction group was treated by injection of 10ml of Dahuang Fuzi decoctionon into the fistula respectively. Blood was collected from heart to detect serum amvlase and endotoxin (ET) levels before the rat being executed. The lung histopathologic changes, pulmonary injury scores and wet/dry weight(W/D) ratios were observed after the rats were executed. The alveolar liquid clearance rate(ALCR), total lung water content (TLW), extravascular lung water content(EVLW) and alveolar epithelial permeability (AEP) were examined in 3,6, 12,24 h after injury.Results There was continuous increase of AEP,TLW and EVLW,as well as progressive reduction of ALCR compared with sham operation group at 3,6,12,24 h after operation. Compared with SAP-ALI group, there was continuous decrease of AEP,TLW and EVLW, and elevated of ALCR at 3,6,12,24 h after operation.Conclusions Dahuang Fuzi decoction can significantly reduce alveolaur epithelial barrier and degree of lung tissue of SAP-ALI rats by inhibiting the elevation of LPS and inflammation reaction.
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Objective To investigate the protective effects of intra-peritoneal fluid resuscitation on small intestinal mucosa in rats with hemorrhagic shock. Method Fifty Sprague-Dawley (SD) rats were randomly (random number) divided into five groups, namely sham operation group (group I ), hemorrhagic shock group (group Ⅱ ), intra-venous fluid resuscitation group (group Ⅲ ) . intravenous fluid resuscitation plus intra-peritoneal saline resuscitation (group Ⅳ ) and intravenous fluid resuscitation plus intra-peritoneal PD-2 solution resuscitation group (group Ⅴ ). The rats of 5 groups were processed with cannulations of right common carotid artery, right femoral vein and left femoral artery with systemic heparinization. The rat models of hemorrhagic shock were established with modified Wigger' s method by which the blood exsanguinated from left femoral artery. The rats of group Ⅲ were resuscitated with shed blood plus twice equal volume of Ringer's solution after modeling of hemorrhagic shock.The rats of group Ⅳ and group Ⅴ were administered intra-peritoneally with 30 mL saline and 30 mL of 2.5% PD-2 solution, respectively as adjuncts to those used in the group Ⅲ . The specimens of blood and small intestine of rats of all groups were collected 60-120 minutes after modeling and resuscitation. The activity of plasma diamine oxidase (DAO) was determined with chromatometry, the level of plasma D-lactic acid (D-LA) with spectorophotometry and the level of plasma lipopolysaccharide (LPS) with nephelometry. The histopathological and ultrastructure changes of small intestine tissue of rats were observed under light microscope and electronic microscope. Results There were remarkable differences in activity of DAO, and the levels of D-LA and IPS in rats between those ingroup Ⅱ and group I (P <0.01), and between those in group V and groups Ⅱ , Ⅲ or Ⅳ (P <0.05 or P < 0.01) The pathomorphology and ultra-structure of small intestine tissues were severely damaged in group Ⅱ compared with those in group Ⅰ , and those markedly lessened in group V compared with groups Ⅱ , Ⅲ and Ⅳ . Conclusions Intraperitoneal fluid resuscitation with PD-2 solution can significantly protect the integrity of intestinal mucosa and the normal permeability of intestinal wall, and blunts the histopathological changes, and restrains bacterial translocation from gut and reduces the level of plasma endotoxin.