| BackgroundStroke has high incidence, morbidity and mortality and becomes the number one killer for human health and life safety. Research on the stroke therapeutic strategies has made great progress,but there still exist several problems to be overcome, for example, the narrow time window of treatment unsatisfactory clinical efficacy and considerable side effects. Given all these problems,looking for effective new therapies has become an urgent and important scientific problem to be conquered.Oxidative stress response is one of the key pathophysiologic mechansim in the process of the evolution of the ischemic cerebral stroke. As an important network nodes of the cascading waterfalls in cerebral ischemia reperfusion injury, the oxidative stress response could reduce cerebral blood flow and attack biological macromolecules and cause cell death, autophagy and apoptosis.For it has many advantage as economical and practical, simple, high maneuverability and small side effects, the electroacupuncture is easily accepted by patients and has a good prospect of clinical therapy in cerebral arterial thrombosis. Electroacupuncture could reduce the formation of malondialdehyde and activate the body’s antioxidant system, as well as improve the function of mitochondrial respiratory chain and antioxidant ability, consequently play a protective effect in nervous system. Ginsenoside Rd is one of the main extractive of ginseng saponins monomer. Either in vitro or in vivo test had proved that the Ginsenoside Rd has good neuroprotective protective effect on cerebral ischemic injuries. Ginsenoside Rd has antioxidant effect by clear away active oxygen directly, reduce the damage of free radicals on cells and restrain peroxidation effectively in the process of cerebral ischemia. Ginsenoside Rd has unique potential in clinical application because it has the very high fat soluble that lead it through the biological membrane and blood brain barrier, and has a longer plasma half-life.This study was to observe whether electroacupuncture combine with Ginsenoside Rd have neuroprotective function against ischemia-reperfusion injury, in order to provide a new thought and theoretical foundation for treatment of cerebral ischemia and reperfusion injury.Experiment 1 The protect function of electroacupuncture combine with Ginsenoside Rd on focal cerebral ischemia in ratsObjective: To explore neuroprotective effect of electroacupuncture combine with Ginsenoside Rd on focal cerebral ischemia injury in rats.Method: 64 male SD rats were randomly assigned to 8 groups(n=8): MCAO group, EA group, GSRd6 h group, GSRd8 h group, GSRd10 h group,EA+GSRd6h group,EA+GSRd8h group and EA+GSRd10h group. MCAO group: Established MCAO animal model. EA group: After the model is established, electroacupunctured at "baihui point" immediately. EA parameters are fixed, the general EA 30 min, dilatational wave, current intensity 1m A, frequency 2/15 Hz. GSRd6 h group: Give GSRd 50 mg/kg through intraperitoneal injection after ischemia 6h. EA+GSRd6h group: After the model is established, electroacupunctured at "baihui point" immediately. EA parameters are fixed, the general EA 30 min, dilatational wave, current intensity 1m A, frequency 2/15 Hz, then give GSRd 50 mg/kg through intraperitoneal injection after ischemia 6h. GSRd8 h group: Give GSRd 50 mg/kg through intraperitoneal injection after ischemia 8h. EA+GSRd8h group: Afterthe model is established, electroacupunctured at "baihui point" immediately. EA parameters are fixed, the general EA 30 min, dilatational wave, current intensity 1m A, frequency 2/15 Hz, then give GSRd 50 mg/kg through intraperitoneal injection after ischemia 8h. GSRd10 h group: Give GSRd 50 mg/kg through intraperitoneal injection after ischemia 10 h. EA+GSRd10h group: After the model is established, electroacupunctured at "baihui point" immediately. EA parameters are fixed, the general EA 30 min, dilatational wave, current intensity 1m A, frequency 2/15 Hz, then give GSRd 50 mg/kg through intraperitoneal injection after ischemia 10 h. 24 hours later, neurobehavioral scores(NBS) were evaluated before the rats were decapitatd, the brains were then removed and stained with TTC to calculate the brain infarct volumes.Result:1) The neurobehavioral scores of EA +GSRd6h group was obviously higher than that of MCAO group(P < 0.05), while the cerebral infarction volume percentage of EA +GSRd6h group was significantly lower than the MCAO group(P < 0.05). 2) The neurobehavioral scores of EA +GSRd8h group was obviously higher than that of MCAO group(P < 0.05), while the cerebral infarction volume percentage of EA +GSRd8h group was significantly lower than the MCAO group(P < 0.05).Conclusion: The electroacupuncture combine with Ginsenoside Rd has neuroprotective effect during focal cerebral ischemia injury in rats. Electroacupuncture can prolong the treatment time window of Ginsenoside Rd to 8 h after ischemia.Experiment 2 The antioxidant effect of electroacupuncture combine with Ginsenoside Rd on focal cerebral ischemiaObjective: To investigate whether electroacupuncture combine with Ginsenoside Rd could play a neuroprotective role by regulating the antioxidant system.Method: 60 male SD rats were randomly assigned to 5 groups(n=12): Sham group, MCAO group, EA group, GSRd group and EA+GSRd group. Sham group: Separate the right side of common carotid artery, external carotid artery and internal carotid artery. Do not insert line plug blocking the middle cerebral artery. MCAO group: Established MCAO animal model. EA group: After the model is established, electroacupunctured at "baihuipoint" immediately. EA parameters are fixed, the general EA 30 min, dilatational wave, current intensity 1m A, frequency 2/15 Hz. GSRd group: Give GSRd 50 mg/kg through intraperitoneal injection after ischemia 8h. EA+GSRd group: After the model is established, electroacupunctured at "baihui point" immediately. EA parameters are fixed, the general EA 30 min, dilatational wave, current intensity 1m A, frequency 2/15 Hz, then give GSRd 50 mg/kg through intraperitoneal injection after ischemia 8h. Take four rats from each group to detect the content of ROS and MDA in brain by ROS kits and MDA kits after reperfusion 24 h. Take four rats from each group to detect 8-OH-d G after reperfusion 24 h. Remaining 4 rats was taken to detect SOD activity, CAT activity and the content of GSH.Result: The content of ROS, MDA and 8-OH-d G of MCAO group is significantly increased compared with the Sham group(P < 0.05). EA + GSRd group reduced the content of ROS, MDA and 8-OH-d G compared with the MCAO group(P < 0.05). The content of SOD, CAT and GSH in MCAO group is significantly increased compared with the Sham group(P < 0.05). EA + GSRd group reduced the content of SOD, CAT and GSH compared with the MCAO group(P < 0.05).Conclusion: Electroacupuncture combine with Ginsenoside Rd have antioxidant effect during focal cerebral ischemia.Experiment 3 The long-term neuroprotective effect of electroacupuncture combine with Ginsenoside Rd on focal cerebral ischemiaObjective: To explore the long-term neuroprotective effect of electroacupuncture combine with Ginsenoside Rd.Method: 1) The SD rats were randomly divided into sham group, MCAO group and EA+GSRd group. Sham group: Separate the right side of common carotid artery, external carotid artery and internal carotid artery. Do not insert line plug blocking the middle cerebral artery. MCAO group: Established MCAO animal model. EA+GSRd group: After the model is established, electroacupunctured at "baihui point" immediately. EA parameters are fixed, the general EA 30 min, dilatational wave, current intensity 1m A,frequency 2/15 Hz, then give GSRd 50 mg/kg through intraperitoneal injection after ischemia 8h. Each group take three rats to determinate weight, water content and brain index in d1、d3、d7 and d14. 2)The SD rats were randomly divided into MCAO group and EA+GSRd group. MCAO group: Established MCAO animal model. EA+GSRd group: After the model is established, electroacupunctured at "baihui point" immediately. EA parameters are fixed, the general EA 30 min, dilatational wave, current intensity 1m A, frequency 2/15 Hz, then give GSRd 50 mg/kg through intraperitoneal injection after ischemia 8h. Neurobehavioral scores(NBS) were evaluated in d1、d2、d3、d4、d5、d6、d7 and d14, the brains were then removed in d7 and d14 and stained with TTC to calculate the brain infarct volumes.Result: Electroacupuncture combine with Ginsenoside Rd lower water content and brain index. The neurobehavioral scores of EA +GSRd group was obviously higher than that of MCAO group(P < 0.05), and the cerebral infarction volume percentage of EA +GSRd group was significantly lower than that of the MCAO group(P < 0.05).Conclusion: The electroacupuncture combine with Ginsenoside Rd has long-term protective effect on focal cerebral ischemia in rats.Summary1. Electroacupuncture combine with Ginsenoside Rd induced neuroprotection in the rat subjected to transient focal cerebral ischemia injury. 2. Electroacupuncture combine with Ginsenoside Rd have antioxidant effect during focal cerebral ischemia. 3. The electroacupuncture combine with Ginsenoside Rd has long-term protective effect on focal cerebral ischemia in rats. |
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