Study On Prophylaxis Of Sinusoidal Obstruction Syndrome By Ginsenoside Rb1 In Rat Model And Its Signal Transduction Mechanism

PartⅠInfluence of ginsenoside Rb1 on rat of sinusoidal obstruction syndromeObjectiveTo investigate the influence of ginsenoside Rb1 on rats of sinusoidal obstruction syndrome (SOS) and explore the protective mechanism of Rb1 on liver.MethodsSOS rat was established by the administration of monocrotaline(Mct). Forty Male SD rats were randomly divided into six groups. Group A were worked as normal control. Group B were only treated with Mct 160mg/kg.Group C and D received Rb1 15mg/kg/d respectively through intraperitoneal injection and intravenous injection two days before being treated with Mct; Group E and F received Rb1 30mg/kg/d respectively through intraperitoneal injection and intravenous injection two days before being treated with Mct; Group G and H only received ginsenoside Rb1 30mg/kg/d respectively through intraperitoneal injection and intravenous injection for 6 days. Then,all rats were killed. Their acites, the ratio of liver/body weight, serum total bilirubin (Tbil), Valley third transaminase (ALT), millet straw transaminase (AST), was detected by nitric acid deoxidizase assy and the tissues were taken for histological esamination.Results1. Compare with group control, the mean of liver/bady weight ratio raised evidently (P<0.01, 3.71 vs 5.57) and the amount of ascites mean achieved 8.2ml. and the level of Tbil, ALT, AST in group Mct increased manifestly(P<0.05,Control TBil:0.72 umol/l,ALT:32.2U/L, AST:136.6U/L vs Model TBil:22.38umol/l,ALT:1015.4U/L ,AST:653.4U/L). Hematoxylin-eosin–stained sections of livers get from five model rats displayed severe damage , severe coagulative necrosis, like granophyric, destruction of the structure of hepatic lobules, monocytes into the lobuleand sinusoidal hemorrhage.2. Contrast the groups pre-treated with Rb1 with group Mct and group control: (1) Pre-treated with Rb1 could decrease the ratio of liver/ bady weight by 24%-34%,and there is no difference between preventive groups and normal group (P>0.05). (2)Descend the amount of ascites from 8.2ml—the mean of group model—to 0.28ml~ 1.4ml, and there is no difference between preventive groups and normal group (P>0.05) .(3) Compare with group Mct, the group pre-treated with Rb1 15mg/kg ip, serum TBil decreased(P<0.05), and the others were not so obvious. (4) we find that the level of ALT in groups pre-treated with Rb1 and subsequently exposed to Mct were significantly decreased than group Mct(P<0.01), and there is no difference between preventive groups and normal group (P>0.05). (5) The level of AST in group pre-treated with Rb1 15mg/kg iv,compare with group Mct,decreased(P<0.05), and four groups had no difference between preventive groups and normal group (P>0.05). (6) The phathology result was analyzed statistically by chi-square test to compare the difference among the groups. The pathological changes of groups pre-treated with Rb1 15mg/kg iv(group C) and 15mg/kg ip (group D) were light than group Mct(P<0.05), but had difference between the above preventive groups and normal group(P<0.01). The improve of groups pre-treated with Rb1 30mg/kg iv(group E) and 30mg/kg ip (group F) were not so obvious, but had significant difference between the above two preventive groups and normal group(P<0.01).3. we employed principal component analysis (PCA) to aggregate analysis the prophylactic effect of the ginsenoside Rb1. The bigger of composite score ,the greater seriousness. The result displayed that the least score was from the group using ginsenoside Rb1 15mg/kg/d through intraperitoneal injection and the biggest was the group using ginsenoside Rb1 30mg/kg /d through intravenous injection.Conclusion1. The phathology of livers from model group, whose liver/bady weight ratio raised and appeared ascites, and altered liver function, conformed to SOS. We using Mct 160mg/kg built a SOS model successfully.2. Rb1 can inhibited the damage of Mct to liver. Prevention though Rb1 can decreased the ratio of liver/bady weight and descend the amount of ascites, reduced pathological changes.3. Among the four prophylactic groups, the groups using Rb1 15mg/ kg/d were better than the 30mg/kg/d groups. These results suggest that low dose of Rb1 may have a better prevention of SOS in rat.4.The two octreotide had no obvious differences in effects,but intraperitoneal injection was easier than intravenous injection, and be counted as an ideal octreotide. PartⅡThe effect of ginsenoside Rb1 on expression of phosphorylationprotein kinase B of sinusoidal obstruction syndrome in ratsObjectiveTo investigate the regulation of eNOS enzyme activity and NO in synthetic by ginse- noside Rb1, and to explore the influence of ginsenoside Rb1 on PI3K/AKT signal pathway and supply theory evidence for prevention of SOS. MethodsBe based on the design of the first part, we taked the animal specimens of the first part to test. Their level of eNOSmRNA in the livers of rats was assyed by Real- Time PCR, the concentration of NO was detected by nitric acid deoxidizase assay, the phosphorylation of AKT and AKT were detected by western blot.Results1.Compare with group control, the concentration of NO in group Mct decreased 52%(group Mct:14μmol/L,group control:28.89μmol/L, P<0.01) and the expression of eNOSmRNA were significantly decreased 40%,(P<0.05).2.The groups which pre-treated with Rb1,the concentration of NO increased 8~12μmol/L than group Mct(P<0.05).There no difference between preventive group with Rb1 15mg/kg ip and normal group (P>0.05),but the others were not(P<0.05).3. In most groups which pre-treated with Rb1 the expression of eNOSmRNA raised manifestly than group Mct, about 39%~66% (Only Rb1 30mg/kg ip preventive group,P>0.05, the others, P<0.05). There was no difference between preventive groups and normal group (P>0.05)4.Compare with group control, P-AKT/AKT ratio declined in group Mct (P<0.05).5.The preventive groups,P-AKT/AKT ratio ascend than group Mct (P<0.05) . There was no difference between preventive groups and normal group (P>0.05)Conclusion1. Our research by detecting serum NO and eNOSmRNA, confirmed the damage of sinusoidal endothelial cells in early SOS.2. We find that Rb1 can enhance eNOS enzyme expression and promote NO synthesis to protect sinusoidal endothelial cells.3.Ginsenoside Rb1can enhance eNOS enzyme activity and the production of NO. The protection of ginsenoside Rb1 in SOS maybe be interrelated with the PI3K/AKT pathway. Deduction: ginsenoside Rb1 can prophylaxis Mct-induced SOS to some extent through which maybe stimulate the PI3K/AKT pathway and then enhance eNOS enzyme activity and the production of NO by sinusoidal endothelial cells. Therefore,our study supplied a preliminary theoretical and experimental foundation for prevention of Rb1.