| Objective Through studyingthe recovery of neurological deficit in rats after intervention of Shi-da-la-zhi-wan on focal cerebral ischemia-reperfusion model and the expression of MAPK-ERK signaling pathway ERK2 in hippocampus,to explore the mechanism of this recipe in preventing neuronal apoptosis in hippocampus of rats with cerebral ischemia-reperfusion injury.Methods238 SPF cleaning grade SD male rats were randomly divided into 17 groups with 14 rats in each group,which are:blank group,sham operation group,model group,nimodipine group,Shi-da-la-zhi-wan low dose group,Shi-da-la-zhi-wan middle dose group and Shi-da-la-zhi-wan high dose group.In the model group and each drug group,need to prepare the MCAO,a rat model of middle cerebral artery embolism.After 2 hours of cerebral ischemia,performe the reperfusion,and these four groups were further refined into three subgroups,i.e.1,3 and 7 days,according to the time of reperfusion and administration.Except for the blank group,sham operation group and model group,saline was given to the stomach,the other drug groups were given corresponding preparations.The concentration was 1.08g/L in nimodipine group and 1.5g/m L,3.0g/m L and 4.5g/m L in low,medium and high dose groups of Shi-da-la-zhi-wan.The volume of gastric perfusion was 1 m L/100 g in rats,and given gastric perfusion twice a day at the same time.During this period,the recovery degree of nerve function defect in rats was evaluated by using Garcia JH score.After that,samples were collected at three time points of 1d,3d and 7d.The relative expression of ERK2 in hippocampus was detected by TUNEL,immunofluorescence,Western blotting of protein(Western Blot)and reverse transcription-polymerase chain reaction(RT-PCR).Results1.Rats' score of recovery degree of nerve function defect :(1)Comparison between model group and sham operation group,the rat neurological defect recovery score reduces(P<0.01);the drug groups were compared with the model group,the rat neurological defect recovery score all rises(P<0.05 or P<0.01).(2)In each drug group,the high dose groupof Shi-da-la-zhi-wan were compared with the other grug groups,the rat neurological defect recovery score all rises(P < 0.05 or P < 0.01),and the difference is significant.(3)The rat neurological defect recovery score rises with the time of 1,3 and 7 days,and was proportional to the time of prolongation.2.Brain tissue TUNEL results:(1)Comparison between model group and sham operation group,the number of apoptotic cells rises(P<0.01);the drug groups were compared with the model group,the number of apoptotic cells all reduces(P < 0.05 or P < 0.01).(2)In each drug group,the high dose groupof Shi-da-la-zhi-wan were compared with the other grug groups,the number of apoptotic cells all reduces(P< 0.05 or P< 0.01),and the difference is significant.(3)The number of apoptotic cells reduced with the time of 1,3 and 7 days,and was inversely proportional to the time of prolongation.3.ERK2 immunofluorescence results:(1)Comparison between model group and sham operation group,the expression of ERK2rises(P<0.01);the drug groups were compared with the model group,the expression of ERK2 all rises(P<0.05 or P<0.01).(2)In each drug group,the high dose groupof Shi-da-la-zhi-wan were compared with the other grug groups,the expression of ERK2 all rises(P<0.05 or P<0.01),and the difference is significant.(3)The expression of ERK2 rises with the time of 1,3and 7 days,and was proportional to the time of prolongation.4.ERK2 Western blotting of protein(Western Blot)results:(1)Comparison between model group and sham operation group,the expression of ERK2rises(P<0.01);the drug groups were compared with the model group,the expression of ERK2 all rises(P<0.05 or P<0.01).(2)In each drug group,the high dose groupof Shi-da-la-zhi-wan were compared with the other grug groups,the expression of ERK2 all rises(P<0.05 or P<0.01),and the difference is significant.(3)The expression of ERK2 rises with the time of 1,3and 7 days,and was proportional to the time of prolongation.5.ERK2 reverse transcription-polymerase chain reaction(RT-PCR)results:(1)Comparison between model group and sham operation group,the expression of ERK2rises(P<0.01);the drug groups were compared with the model group,the expression of ERK2 all rises(P<0.05 or P<0.01).(2)In each drug group,the high dose groupof Shi-da-la-zhi-wan were compared with the other grug groups,the expression of ERK2 all rises(P<0.05 or P<0.01),and the difference is significant.(3)The expression of ERK2 rises with the time of 1,3and 7 days,and was proportional to the time of prolongation.Conclusions1.Shi-da-la-zhi-wancan repair the defect of nerve function in rats.2.Shi-da-la-zhi-wan can inhibit cell apoptosis in rats with focal cerebral ischemia-reperfusion injury,this may be related to the regulation of ERK2 expression in MAPK-ERK signaling pathway. |
PDF Full Text Request