Effects Of Solium Ginkgo Extraction On Endovascular Therapy For Ischemic Cerebral Disease And The Study Of Response Surface Optimized Extraction Of Carbohydrate Compound From Folium Ginkgo Plus Its Protective Effect In I/R Animal Model

Stroke was the fourth most common cause of death worldwide in2008inthe United States, esulting in5.7million deaths (~10%of the total). In China,cardiovascular disease is the first leading cause of death. On average, every45seconds, there is a stroke victim in the United States; every3minutes,someone dies from stroke. The data is similar in China, for the cardiovasculardisease, every12seconds, there is a new suffer; every21seconds, someonedies from it. The above facts reflect the urgency of preventing stroke anddiscoveing new drugs/method to treat stroke.Ischemic stroke accounts for about87%of strokes, it results from theocclusion of cerebral vascular vessels, including the common branches of theinternal carotid arteries and the basilar artery and its branches. Brain tissueceases to function if deprived of oxygen for more than60to90seconds, andafter approximately three hours will suffer irreversible injury possibly leadingto death of the tissue, i.e., infarction. This is why fibrinolytics such asalteplase are given only until three hours since the onset of the stroke. Ifvasodilation failed to increase blood flow, Cerebral Perfusion Pressure (CPP)decreases further. At last neurological deficit will occur. Middle cerebralartery occlusion (MCAO) usually causes contralateral hemiplegia. It is easilyto copy with an animal model at experiment.Atherosclerosis may disrupt the blood supply by narrowing the lumen ofblood vessels leading to a reduction of blood flow, by causing the formation ofblood clots within the vessel, or by releasing showers of small emboli throughthe disintegration of atherosclerotic plaques. The ischemia area is referred toas the "ischemic penumbra". Since blood vessels in the brain are now occluded, the brain becomes lack of energy; the treatment strategy is to restorethe energy supply by changing the fluidness of blood or openning the arteryagain quickly. Definitive therapy is aimed at removing the blockage bybreaking the clot down (thrombolysis), or by removing it mechanically(thrombectomy). The more rapidly blood flow is restored to the brain, thefewer brain cells die. For the drug treatment, only treatment with medicationssuch as aspirin, clopidogrel and dipyridamole may be given to preventplatelets from aggregating. Other medication such as Folium Ginkgo is notapproved by FDA even for prevention stroke, although it has been used inChina for more than2000years. But more and more studies have been done toprove the useness of Folium Ginkgo to use as a supplementary medication.This study is focus on the effection of Folium Ginkgo for clinical patients andthe new extractions for animal experiment. There are several new discoveriesfrom the study.Part Ⅰ The effect of Folium Ginkgo extraction on mechanicalthrombectomy based on clot intensity at ICA occlusion.Objective: To evaluate human acute stroke cases to verify if thepre-treatment signal intensity of the clots in the occluded internal carotidartery (ICA) could affect the degree of angiographic recanalization afterendovascular treatment; and to analyze the clinic cahracterastic of FoliumGinkgo consumer patients, to evaluate the effect of the hereb on theendovascular therapy.Methods: We retrospectively reviewed74consecutive patients with ICAocclusion diagnosed with MRI and MRA before endovascular treatment inUCLA stroke center between2002and2012. Clot FLAIR intensity in the ICAterminus and recanalization score in DSA were collected, and re-assessedparticularly and blindly, the intensity was assessed with Image J software.Characteristics were compared between the successful reperfusion group(TICI scale2b-3) and without successful reperfusion group (TICI scale1-2a)after endovascular treatment, and multivariate logistic regression analysis ofperfusion was conducted with SPSS software. Results: Successful reperfusion was achieved in40.3%of patients(27/67), and the clots in the reperfusion cases showed significantly higherFLAIR intensity (P=0.020). More senile patients were achieved successfulreperfusion with the mean age of74years old, the differences is significant(P=0.038). The patients as Folium Ginkgo consumer achievd less successfulreperfusion compared those who didn’t consume Folium Ginkgo, withsignificant differences (11%vs.35%, P=0.044). In multivariate logisticregression analysis, only high FLAIR clot intensity was associated withfavorable reperfusion (odds ratio,0.05;95%confidence interval,0–0.68; P=0.025).Summary: FLAIR clot density at ICA occlusion in patients of acutestroke could independently predict successful reperfusion. A clinicalapplication of FLAIR clot imaging may be useful to predict the treatmentoutcome in ICA occlusion. Administrated Folium Ginkgo leave extract canimprove one’s blood flow by increasing red blood cell deformability anddecreasing red cell aggregation, lessen the chance suffered fromerythrocyte-rich clots embolus. We should put this phenomeno intoconsideration when we made a decision to choose adaptive treatment methodsor devices.Part Ⅱ The effect of Folium Ginkgo extraction on the individual PTASfor intracranial vertebral and Basilar artery stenosisObjective: High periprocedural complication rate is one of thelimitations of endovascular treatment despite greatly reducing the risk of therecurrent stroke. Based on SAMMPPRIS trial but with the modification fortargeted population and individual angioplasty method, we want prospectivelyto evaluate the efficacy and safety of the individual angioplasty method forpatients with intracranial atherosclerotic disease (ICAD), and to evaluate theeffect of Folium Ginkgo extraction on this group of patients.Methods: Between November2011and November2012,122patientswere consecutively recruited into this prospective single-center study.Individual PTAS was a new method for intracrinal artery stenosis treatment. Based on this method, all patients were divided into3groups based on4angioplasty methods: Apollo (A) Group, Gateway/Gateway+Apollo (G-GA)group and Gateway+Wingspan (GW) Group. The primary end points weresuccessful procedure rate and any vascular event at discharge days. The cliniccharacteristic and the primary end points were evaluated based the FoliumGinkgo administration.Results: Overall Technical Success Rate was97.5%(151/156).Intracranial stenting was successful in97.6%of patients in A group,96.8%inGW group and only90.9%in G-GA group with significant differencesbetween three groups (P=0.045). The average residual stenosis of all patientswas11.84%. Residual stenosis rate in G-GA group was higher (16.35±15.52%)than those in other groups, and the A group got the lowest residual stenosisrate of (9.42±8.81%, P=0.009). The patients in Folium Ginkgo group havelower Hs-CRP and higher NIHSS, compared with that in No Folium Ginkgogroup, the differences are significant respectively (2.20±2.23mg/L vs.4.54±3.75mg/L, P=0.012;1.47±2.44vs.0.54±1.18, P=0.006). And moremRS=1or2patients are in Folium Ginkgo group, but more mRS=0patientsare in No Folium Ginkgo group (P=0.047). More patients in No FoliumGinkgo group are suffered with TIA, the patients in Folium Ginkgo group aretended to suffer stroke, the differences are significant (P=0.047). But there areno significant differences in the primary endpoints and other clinicalcharacteristic between the two groups.Summary: Individual angioplasty method for ICAD treatment according tolesion morphology was feasible and safe in patients with poor collateralcirculation. Further follow-up is needed to assess the long-term efficacy of theindividual procedure and multicenter study will be planned to confirm it.Folium Ginkgo can less the TIA onset and resist erythrocyte-rich clots formatby the maner of anti-inflammatory and increasing red blood cell deformability. Part Ⅲ Response Surface Optimized extraction of carbohydrateCompound from Folium Ginkgo and its protective effection for I/RanimalObjective: To investigate new extraction method for Folium Ginkgo, andfurther study the protective effect of carbohydrate extraction for MCAocclusion I/R mice model.Methods: Response surface methodology (RSM) was used to extractcarbohydrate extraction from Folium Ginkgo. Male Wister rats were used toestablish Middle cerebral artery occlusion (MCAO) animal model, wichi is anoxidative injury in focal cerebral ischemia–reperfusion animal model. Malewistar rats were randomly divided into5groups: sham control group, I/Rgroup, Medcine I group (200mg/kg/day carbohydrate extraction), Medcine IIgroup (400mg/kg/day carbohydrate extraction), and Medcine III group(600mg/kg/day carbohydrate extraction). Rats in sham control group and I/Rgroup were administered by gavage with salidroside for30days. Rats inmedcine group were administered according herb extraction for30days. Theinfarction volume was calculated with image analyzer by TTC staining. Theactivities of superoxide dismutase (SOD), CAT and GSH-Px and the levels ofMDA, Caspase-3and NO in brain tissue were determined by separatebiochemical kit. And the protein of iNOS and COX-2were detected bywestern blot. The cDNA of iNOS and COX-2were detected with RT-PCR.Data were presented as xˉ±s and analyzed with ANOVA and LSD using SPSSstatistical program. A level of P＜0.05was considered statistically significant.Results:(1) With RSM we got2fractions from Folium Ginkgo,Fraction-I was a heteropolysaccharide and consisted of L-rhamnose,L-arabinose, D-mannose, D-glucose, and D-galactose, with the molar ratio of1:1.7:2.2:1.8:7.0. Fraction-II also was a heteropolysaccharide and consisted ofL-rhamnose, D-mannose, D-glucose and D-galactose, with the molar ratio of1.8:2.1:1.4:4.3.(2) I/R animals displayed a significantly decrease of theactivities of SOD, CAT, GSH-Px, NO and increase the levels of MDA andCapspase-3in brain tissue compared with the sham control group (P<0.01).The activities of SOD, CAT, GSH-Px and NO were significantly increasedafter treated Folium Ginkgo extraction compared with the I/R group and shamcontrol group (P<0.01, P<0.01). On the contrast, MDA and Caspase-3were significantly decreased after treated Folium Ginkgo extraction compared withthe I/R group and sham control group (P<0.01, P<0.01).(3) The cDNA andprotein of iNOS and COX had the similar tendency with SOD and NO,compared with sham control group iNOS and COX-2were increased in theI/R group, and were decreased after treated with Folium Ginkgo extraction inthe medicine group (P<0.001).Summary: Carbohydrate compound also have the cerebral protectivebioactivities and they can reduce oxidative injury and cause reduction ofinfarct volume following cerebral artery ischemia and reperfusion injury. Inthis respect carbohydrate extraction from Folium Ginkgo might be the choiceof therapeutic agent for patients with focal cerebral ischemia.Conclusions: In summary, our study found that Folium Ginkgo canreduce erythrocyte-rich clot formation by influence the red blooddeformability and other ways for the patients with ischemic stroke of severestenosis intracranial artery, thus affecting the manner of stroke onset andtreatment. Carbohydrate extract of Ginkgo biloba also has a stronganti-inflammatory, anti-oxidation, anti-stress activities and can improve theblood supply of the ischemic area further reduce the ischemic area of the I/Ranimial. The study gives us some new idea or alternative option for preventingand treating ischemic cerebrovascular disease with Folium Ginkgo extraction.