| PartⅠInhibitory effect of Isoliensinine, Liensinine, Neferine on Cholinesterase of plasma in ratIn this study, the effect of IL, Lien and Nef on Cholinesterase (ChE) were investigated using colorimetric determination. It was found that Isoliensinine, Liensinine and Neferine could inhibit ChE in a concentration-dependent manner. In the acetylcholinesterase inhibitory assay, the IC50 of AchE induced by IL, Lien and Nef were 6.82±0.25μM, 7.17±0.46μM, 7.64±0.82μM, respectively. In the butyrocholinesterase assay, the IC50 of BchE induced by IL, Lien and Nef were 5.48±0.50μM, 6.84±0.53μM, 15.51±2.20μM, respectively. The IC50 of AchE and BchE induced by Galanthamine were 0.41±0.03μM and 3.7±0.11μM. The selectivity index (SI) value of IL, Lien, Nef and Gala on ChE were 0.80, 0.95, 2.03, respectively. The results above demonstrated that all three compounds (IL, Lien and Nef) have the ability of non-selective inbition on both AchE and BchE.PartⅡIsoliensinine, Liensinine and Neferine improve scopolamine-induced memory impairment in miceTo evaluate the effects of IL, Lien and Nef on Scopolamine induced memory impairment in mice, Morris water maze task and Y maze task were taken to measure the reference memory and short-term memory, respectively. In Morris water maze task, the latency to first target– site crossover of Saline group, Scop, Gala + Scop group, Nef (1, 10, 30 mg/Kg) + Scop group, IL (1, 10, 30 mg/Kg) + Scop group, Lien (1, 10, 30 mg/Kg) +Scop group were 12.81±2.35 s,20.59±1.40 s,11.83±2.63 s,16.32±2.92 s,15.83±2.56 s,14.82±1.91 s,16.54±2.72 s,16.43±2.06 s,14.45±2.47 s,17.79±2.38 s,14.97 ±2.06 s,12.99±1.22 s, respectively. It was shown that Neferine (10, 30 mg/Kg), Isoliensinine (30 mg/Kg) and Liensinine (10, 30 mg/Kg) has the therapeutic efficacy compared with the Scop group. In the Y maze task, the alternation rate of Saline group, Scop group, Gala + Scop group, Nef(10 mg/Kg) + Scop group, IL(30 mg/Kg) + Scop group, Lien(10mg/Kg) + Scop group were 43.98±2.31%,34.67±1.96%,46.34±2.87%,41.97±2.18%,44.74±2.23%,41.23±2.67%, respectively. Compared with Scop group, it was found that IL(30 mg/Kg), Lien (10 mg/Kg) and Nef(10mg/Kg) were able to rescued the short-term memory in Kunming mice which was co-treated with scopolamine 30 minutes before test. So we can conclude that IL, Lien and Nef has the anti-amnesic activity.PartⅢIsoliensinine, Liensinine and Neferine inhibit Cholinesterase in rat's cerebral cortexTo determine the anti-cholinesterase properties of IL, Lien and Neferine in vivo, spectrophotometry was taken to determine the ChE activity in rat's cerebral cortex. IL (30mg/Kg), Lien (10mg/Kg) Nef (10mg/Kg) and Gala (0.5mg/Kg) was given by intragastric administration, twice daily for five consecutive days. Thirty minutes later after the last administration, the rat was sacrificed and the cerebral cortex were isolated for the ChE activity determination. The inhibition rate (%) of ChE induced by IL, Lien, Nef and Gala was calculated by comparison with the enzyme activity obtained from rats which was administrated Saline. The protein level of each group's 1% soliquoid was also determined by colorimetric determination using Coomassie brilliant blue as the developer. The protein level of Saline group, Gala group, IL group, Lien group and Nef group were 1.09±0.06μg/ml, 1.09±0.05μg/ml, 1.13±0.04μg/ml, 1.05±0.02μg/ml, 1.09±0.05μg/ml, respectively. we observed that there were no significance differences of protein level between groups. The AchE activity of Saline group, Nef group, IL group, Lien group and Gala were 1.23±0.10U/μgprot, 0.87±0.04U/μgprot, 0.93±0.04U/μgprot, 0.87±0.04U/μgprot, 0.67±0.05 U/μgprot, respectively. The inhibition rate (%) of Neferine group, IL group, Lien group and Gala group on AchE were 29.17±3.35 %, 24.88±3.17 %, 29.70±2.59 %, 45.61±4.61 %. The BchE activity of Saline group, Nef group, IL group, Lien group and Gala group were 0.61±0.03U/μgprot, 0.49±0.02U/μgprot, 0.53±0.01U/μgprot, 0.50±0.01U/μgprot, 0.55±0.01U/μgprot, respectively. The inhibition rate of Neferine group, IL group, Lien group and Gala group on BchE were 18.68±2.62%, 12.85±2.29 %, 17.00±1.17 %, 9.06±1.89%.Conclusion: (1) Isoliensinine, Liensinine and Neferine could inhibit ChEs in a concentration-dependent manner. (2) Neferine(10mg/Kg, 30mg/Kg), Isoliensinine(30mg/Kg) and Liensinine(10mg/Kg, 30 mg/Kg) could antagonizes the amnesic effect of scopolamine in Morris water maze task. And Isoliensinine (30 mg/Kg), Liensinine (10 mg/Kg) and Neferine (10mg/Kg) were able to rescued the short-term memory in Y maze task. (3) Isoliensinine, Liensinine and Neferine could penetrate the blood-brain barrier and inhibits ChE in rat's cerebral cortex. So we could conclude that Isoliensinine, Liensinine and Neferine has the ability of anti- amnesic effect by ChE inhibition. |
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