The Effect And Mechnism Of Curcumin Analogues Dispersion On Mice With Learning And Memory Disorders Induced By Scopolamine

Objective:1.To observe the effect of Curcumin analogues on the mice with learning and memory impairment induced by scopolamine,and to explore its mechanism.2.To observe the effect of Tetrahydrocurcumin(THC)solid dispersion on scopolamine induced learning and memory impairment in mice,and the effect of H2O2 oxidative damage on PC 12 cells model was further explored to research its mechanism on acetylcholine pathway,antioxidant and anti-apoptosis.Methods:1.Curcumin analogues were used to improve learning and memory impairmen:a model of learning and memory impairment induced by scopolamine was used in mice.The mice were fed with Curcumin analogues:Curcumin(200 mg/kg),Demethoxycurcumin(200 mg/kg),Bis Demethoxycurcumin(200 mg/kg)and Tetrahydrocurcumin(200 mg/kg).①Effects of Curcumin analogues on mice with learning and memory impairment(Morris water maze)were observed.② Pathological changes of curcumin on brain tissues in mice with learning and memory impairment were observed.③Effects of curcumin on contents of ChAT,AChE and ACh in brain tissues of mice with learning and memory impairment(Immunohistochemistry)were studied.2.The effect of Tetrahydrocurcumin(THC)on the learning and memory impairment in mice induced by scopolamine was investigated:the same animal model was established,the mice were fed with Tetrahydrocurcumin(THC):high dose(400 mg/kg),medium dose(200 mg/kg)and low dose(100 mg/kg),respectively.① The effect of Tetrahydrocurcumin(THC)on learning and memory was investigated by water maze test.②Observation of Nissl bodies in brain tissues of mice was performed by Nissl staining.③ Effects of Tetrahydrocurcumin(THC)on contents of ChAT,AChE and ACh in brain tissues were investigated by the immunohistochemistry method.④Influences of Tetrahydrocurcumin(THC)on contents of GSH-Px,MAD,SOD in hippocampus were studied by ELISA.3.The effect of Tetrahydrocurcumin(THC)on apoptosis of PC 12 cells induced by H2O2 was investigated.① The activity of Tetrahydrocurcumin on PC12 cells was determinated by CCK-8.PI and Hoechst staining was performed for apoptosis research.② ROS staining was used to detect changes of reactive oxygen species.Results:1.Curcumin analogues were used to improve learning and memory impairment:① Compared with the model group,in the training trial of water maze test,curcumin(200 mg/kg)and bisdemethoxycurcumin(200 mg/kg)shortened the escape time(P<0.05)of mice and reduced the total movement distance of mice(P<0.01).Tetrahydrocurcumin(200 mg/kg)significantly shortened the escape time(P<0.01)and reduced the total movement distance of mice(P<0.01).In the spatial probe trial,curcumin(200 mg/kg)and Bisdemethoxycurcumin(200 mg/kg)significantly increased the escape time(P<0.01),and the number of mice crossing the platform area was significantly increased(P<0.01).Mice treated with tetrahydrocurcumin(THC,200 mg/kg)moved longer distances and more frequently crossed the platform area(P<0.01).Curcumin analogues improved the ability of spatial learning and memory in mice.② There was no significant difference between the treatment group and the model group.③ The immunohistochemical observation of mice brains showed curcumin-like compounds increased ChAT protein content in brains of mice:Curcumin(P<0.05),Demethoxycurcumin(P<0.05),Bisdemethoxycurcumin(P<0.05)and Tetrahydrocurcumin(P<0.01).At the same time,AChE protein content in brains of mice was reduced:Curcumin(P<0.05),Demethoxycurcumin(P<0.01),Bisdemethoxycurcumin(P<0.05).2.The effect of Tetrahydrocurcumin solid dispersant on scopolamine induced learning and memory impairment in mice:① Different doses of Tetrahydrocurcumin all reduced the escape time(P<0.01)and reduced the total distance of movement(P<0.01)in the training trial.Spatial probe trial showed that Tetrahydrocurcumin increased the movement time and distance(P>0.05)in the effective quadrant.② The number of Nissl bodies in T400(THC 400 mg/kg)group was more than that in the model group.There was no significant difference between groups.③ High dose(400 mg/kg)of Tetrahydrocurcumin solid dispersant increased ChAT and reduced AChE protein contents(P<0.05).④ And THC reduced the content of MDA in the hippocampus(P<0.01),and increased the content of SOD(P<0.05).It showed that Tetrahydrocurcumin not only regulated the synthesis of acetylcholine,but also had the antioxidant effect.3.The effect of Tetrahydrocurcumin on H2O2 induced PC 12 cells injury:the survival rate of PC 12 cells was significantly decreased in CCK-8 test when the concentration of hydrogen peroxide was 100 umol/L(P<0.01).① PI and Hoechst staining showed that there was no significant difference between H2O2+THC group and H2O2 group(P>0.05).② The level of reactive oxygen species(ROS)detected by ROS staining in H2O2+THC group was significantly lower than that in H2O2 group(P<0.01).Conclusion:1.Curcumin analogues can significantly improve the learning and memory difficulties induced by scopolamine,the mechanism may be by regulating ChAT and AChE.2.Tetrahydrocurcumin solid dispersion can improve the memory ability of mice,the mechanism may be related to the combined action of regulating acetylcholine pathway and anti-oxidation.3.Tetrahydrocurcumin inhibites the increase of reactive oxygen species induced by H2O2 on PC 12 cells.