| Objective:To study the protective mechanism of ligustrazine (TMP) on kidney injury of rats with severe acute pancreatitis(SAP) by detecting the level of SOD, MDA, TNF-a, BUN, Cr,AMY in serum and observing the expression of Bax, Bcl-2 in the tissue of pancreae and kidney. Method:45 female SD rats with the weight of 200-250g, were randomly divided into normal group (N group), severe pancreatitis group (P group), TMP treated group (C group). Each group was randomly divided into three subgroups according the time points of 12h,24h,48h. Rats were anesthetized by intraperitoneal injection, exposure to the pancreas, take the pancreatic duct as anatomical location mark to find import duodenal papilla. Normal group were only taken two kinds of invasive operation of celiotomy and stretching the intestinal wall, Saline (10ml/kg) was injected from the rat tail vein in severe pancreatitis group. TMP (200mg/kg) was injected from the rat tail vein in TMP treated group. The serum superoxide dismutase and malondidehyde were tested by use SOD radioactive 1251-radioimmunoassay kit and MDA thiobarbituric acid (TBA method) kit. The level of BUN, Cr, AMY were tested by use Automatic biochemical analyzer; apoptotic index of kidney was detected by TUNEL; the expression of Bcl-2 and Bax protein in the renal was detected by immunohistochemistry and Western blot. All results were analysis by use SPASS16.0. Results:1. Pathologic changes of the pancreas:Peritoneal exudate, pancreatic edema,adhesion of pancreatic surrounding tissue were not abserved in the N group. P group shows a small amount of bloody ascites, pancreatic color purple, associated with congestion,edema and local necrosis and bleeding, with a little fat saponification around the greater omentum, mesentery, pancreas at 12h; more red bloody ascites were find at 24h, the color of the pancreas was dark brown, and the pancreas was adhered to the surrounding tissue, more fat saponification were observed in the omentum, mesentery and Peripancreatic tissue; a lot of dark red turbid ascites were observed at 48h, pancreas complicated with massive necrosis, hemorrhage, and large areas of fat saponification were brown. At the TMP treatment group, the bloody exudate were also abserved. However, the amount of ascites compared with the severe pancreatitis group was significantly little. Congestion and edema in the pancreas can also be found, but the range was significantly reduced compared with severe acute pancreatitis. Observation under light microscope: in normal group, the structure of pancreatic tissues and cells were integrity, and there were no inflammatory cells surrounding interstitial infiltration. In severe acute pancreatitis group, Pancreatic lobules and septal were widening at 12h, and accompanied by a small amount of inflammatory cell infiltration and focal necrosis, hemorrhage; at 24h, the degree of pancreatic damage was increased than the degree at 12h, Pancreatic lobules and septa widened further, accompanied by the amount of interstitial cell infiltration, by acinus cells swelled and partial pancreatic acinar necrosis, hemorrhage. At 48h, the pancreas was serious damaged, showing severe swelling of acinar cells, large areas of necrosis, hemorrhage, part of the lobular structures are completely illegible. TMP treatment group, the degree of necrosis, swelling in pancreatic, peripancreatic infiltration of mesenchymal cells and other indicators were reduced than the severe acute pancreatitis at the same time point.2. The pathological changes of kidney: there were no congestion, edema and bleeding in the normal group watch with naked eye, capsule no tension, no exudate perirenal space, perirenal fat without saponification. Severe pancreatitis group, the appearance of kidney was mild edema, the volume increased slightly, the color of kidney was dark brown at 12h; at 24h, edema in kidney heavier than before, and bleeding points scattered; at 48h, congestion and edema of kidney were continued to increase, color of kidney was purple-brown, showing that a large number of bleeding, capsule tense, part of the perirenal fat saponification. TMP treatment group, the kidney gradually over time and then bleeding the formation of edema, exudate perirenal space began to emerge. But the degree was lesser than the severe acute pancreatitis group at the same time point. Observation through light microscope: Normal group, Rat kidneys were either no glomerular or tubular edema and there were no infiltration of inflammatory cells kidney tissues. Severe pancreatitis group, both glomeruli and tubules were involved at 12h, manifested as congestion, swelling and the tubular was more obvious, infiltration of a little inflammatory cell were observed in kidney tissues. At 48h the swelling of the kidneys can not identify the part of the tubular structure, we can see more of renal interstitial inflammatory cells. With time going, there has been necrotic epithelial cells, inflammatory cells, and the tube. TMP treatment group over time periods kidney tubular epithelial cells appeared necrotic, inflammatory cell infiltration of renal tissues, such as the performance of perirenal fat saponification, but its time injury with severe acute pancreatitis group were reduced. 3. Biochemical Indicators:Severe pancreatitis group at different time points of serum BUN, Cr, AMY, TNF-a levels at all time points compared with the normal group was significantly higher(P<0.05). TMP treatment group at all time points of serum BUN, Cr, AMY, TNF-a levels compared with the the severe pancreatitis group was lower (P<0.05).4.MDA, SOD. Severe pancreatitis group at different time points of serum MDA levels compared with the normal group was significantly higher(P<0.05). Severe pancreatitis group at different time points of serum SOD levels compared with the normal group was significantly lower (P<0.05). TMP treatment group at all time points of serum MDA levels compared with the the severe pancreatitis group was lower (P<0.05). TMP treatment group at all time points of serum SOD levels compared with the the severe pancreatitis group was higher (P<0.05).5. Kidney pathology score:severe pancreatitis group renal pathology score was increased gradually progress over time, TMP treatment group at all time points of Kidney pathology score compared with the the severe pancreatitis group was lower (P<0.05).6. Bcl-2/Bax ratio:Severe pancreatitis group at different time points of Bcl-2/Bax ratio levels compared with the normal group was significantly lower (P<0.05). TMP treatment group at all time points of Bcl-2/Bax ratio levels compared with the the severe pancreatitis group was higher (P<0.05). The results showed that TMP can increase Bcl-2/Bax ratio at each time point. Conclusion:SAP TMP can protect renal injury.The mechanism may be through scavenging oxygen free radicals, regulation of apoptosis and the release of inflammatory mediators and so on to achieve. |
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