Gynostemma Saponins To Mitigate The Effect Of Ischemia Reperfusion Injury In Rats And Mcpip Expression In Cerebral Ischemia Injury

Objective:To investigate the effects and mechanisms of Gypenosides (GPs) on cerebral ischemia and reperfusion injury in rats.Method:80SD rats were randomly divided into five groups:①Sham operation group: double distilled water (DDW,10ml/kg/d);②Model group:DDW (10ml/kg/d);③GPs low-dose group:100mg/kg/d;④GPs high-dose group:200mg/kg/d;⑤Positive control group:nimodipine group (20mg/kg/d,) n=16. Rat cerebral ischemia-reperfusion model was built by Zea Longa method. Each group of rats were anesthetized and treated with cerebral ischemia-reperfusion, respectively. Until the rats awake, the rats were divided according to the eurological deficit scores and administered orally for7days. The eurological deficit score was detected everyday. Seven days later, sections were stained by TCC and detected volume of infarct. Change of pathological in the brain tissue was detected by HE staining; the expression of IL-1β, MCP-1and NF-κBp65nuclear translocationrat, in cerebral cortical, were detected by immunohistochemistry.Result:1. Neurological deficit score:no neurological deficit in sham operation group, the highest neurological score turned up after cerebral ischemia-reperfusion first day in the model group and then gradually reduce; Compared with model group, GPs high and low dose groups were found lower neurological scores was in the third day and have effect of dose-dependent.2. Infarct volume:Infarction lesions were not found in the sham operation group. A large range of ischemia was found in model group; Infarction lesions in GPs high and low dose groups were smaller than control group significantly (P<0.01); There was no significant difference between the GPs high and low groups (all P>0.05).3. Changes of Histopathological:Cortex of sham operation group without ischemic lesions histopathological changes. Most of the nerve cells of the brain tissue of model group atrophy, degeneration, or necrosis, cytoplasm degeneration, porosity, nucleus deeply stained with varying degrees of condensation, dissolution, irregular collapse of the large number of nerve fibers, the structure disappeared and edema of glial cells. There are different degrees of reduction of GPs high and low groups compared with model group.4. IL-iβ, MCP-1and NF-KBp65nuclear translocation expressed in cortical tissue:There are few expressions of IL-1β and NF-κB p65nuclear translocation in the sham operation group, but without MCP-1expression; Compare with the sham operation group, IL-1(3, MCP-1expressions and NF-KBp65nuclear translocation significantly increased in the model group.(all P<0.01); compare with the model group, GPs high and low dose groups, IL-1β and MCP-1expressions decreased, but nuclear translocation of NF-KBp65reduced significantly(P<0.01).Conclusion:①Ps can reduce neurological damage, infarct volume and histopathological changes of brain;②GPs reducing the cerebral ischemia-reperfusion injury in rats which were relative to inhibit of inflammatory molecules IL-1β, MCP-1expression and regulate of these two molecules expression of the transcription factor NF-κB p65activity Objective:To observe the MCPIP expression with ischemia time in the injured tissues of the rat lack of cerebral blood.Method:50SD rats were randomly divided into five groups:①Normal control group;②schemia for6h group;③schemia for12h group;④schemia for24h group;⑤schemia for48h group (all n=10). The damage model of middle cerebral artery ischemia (MACO) was built by suture method. When animals, in each group, awaked and given a score of neurological behavior. By using HE stain to observe pathological changes of brain tissue; expression of MCP-1mRNA and MCPIP mRNA in cortex ischemia were analysed at different time by real-time fluorescence quantitative PCR (Real-Time PCR). The MCPIP protein expression in cortex ischemia was detected by Western blot at different time.Result:1. Brain histopathological results:in normal control group, cortical tissue was dense structure, nerve cells morphology clear, a large number of cells, arrangement neat, complete nuclear membrane and nucleolus clear; Ischemic for6-12hours, brain cells swelled, the cells surrounding have gap, nucleus deep stained, nuclear membrane shrinked and unclear cytoplasmic boundaries. Ischemia for24hours, nerve cells around the gap expanded, increasing the degree of cells swelling, nuclear membranes shrinkage was more serious and most of the nuclear lysed. Ischemia for48hours, the nerve cells apoptosis compared with ischemia for24hours significantly reduced, and the degree of nuclear membranes shrinkage were reduced.2. Real-time PCR results showed that no expression of MCP-1mRNA in normal control group, but with a few expression of MCPIP mRNA. Ischemia group MCP-1mRNA and MCPIP mRNA expression were significantly higher than the normal control group. Compared with model group, MCP-1mRNA and MCPIP mRNA expression in Cortex for48hours higher than in Cortex for24hours (P<0.01).3. Western Blotting results:There are a small amount of expressions of MCPIP protein in normal control group, however, in model group, expressions of MCPIP protein in cortex with different time of ischemia are significantly higher than normal control group, the most expressions for48hours (P<0.01)Conclusion:MCPIP expressions gradually increased with time-dependent in ischemia injury of rat brain tissue.