Objective:To determine the chemical structure of the new compound and investigate the protective effects of Tinosporaic acid A and B towards in-vitro neuro.Methods:The structures of two new compounds were established ...Objective:To determine the chemical structure of the new compound and investigate the protective effects of Tinosporaic acid A and B towards in-vitro neuro.Methods:The structures of two new compounds were established by analyzing its 1D and 2D NMR spectra as well as HRESIMS.Their neuroprotective effects with respect to the antioxidant properties were evaluated by radical scavenging tests and hydrogen peroxide-injured oxidative stress model in PC12 cell lines.Cell morphology of treated PC12 cells was observed by phase contrast microscopy.In-vitro MTT assay,lactate dehydrogenase activity assay and oxidative stress markers(intracellular ROS production,MDA level,and caspase-3 activity) were used to evaluate the protective effects against hydrogen peroxide induced cytotoxicity in PC12 cells.Results:The two new compounds,named Tinosporaic acid A and B,were isolated and identified from the stem bark of Tinospora hainanensis.Cell viability studies identified a representative concentration for each extract that was subsequently used to measure oxidative stress markers.Both extracts were able to reverse the oxidative damage caused by hydrogen peroxide,thus promoting PC12 cells survival.The concentration of Tinosporaic acid A and B were 86.34 μg/mL and 22.06 μg/mL respectively,which is neuroprotective for EC_(50).The results indicated that both of them significantly attenuated hydrogen peroxide-induced neurotoxicity.Conclusion:The two new compounds isolated from ethanol extracts of Tinospora hainanensis are the promising natural ones with neuroprotective activity and needed for further research.展开更多
Objective:To explore the anti-gout effects of four traditional Chinese medicine extracts of Eucommia,Sargentodoxa cuneata,Caulis spatholobi and Phellinus igniarius.Methods:Monosodium Urate(MSU)was used to establish a ...Objective:To explore the anti-gout effects of four traditional Chinese medicine extracts of Eucommia,Sargentodoxa cuneata,Caulis spatholobi and Phellinus igniarius.Methods:Monosodium Urate(MSU)was used to establish a mouse gouty arritis model.With coloricine as the positive control group,the effects of four traditional Chinese medicine extracts of Ecuador,Sargentodoxa,Caulis spatholobi and Phellinus igniarius on rice were studied;The hot plate method was used to observe the analytical effects of the four groups of traditional Chinese medicine extracts;the enzyme-linked immunoassay(ELISA)was used to detect residual interference-1 beta(IL-1 beta);the four traditional Chinese medicine extracts were simultaneously determined on the white blood cells,and monocytes in the blood of mice as well as the influence of neutrophils and histopathological changes of toes.Results:Compared with the normal group,the joint diameter ratio,blood leukocytes,monocytes,neutrophils,IL-1βat different time points in the model group were significantly increased(P<0.01),and the pain percentage was significantly reduced(P<0.05).Compared with the model group,the joint diameter ratio of the colchicine group,the Eucommia group,the Sargentodoxa group,the Caulis spatholobi group and the Phellinus sibiricum group was significantly reduced(P<0.05),and the pain percentage was significantly increased(P(0.05).The four groups of traditional Chinese medicine extracts significantly decreased the white blood cells,monocytes,and neutrophils in the blood of mice(P<0.05),and the IL-1βexpression levels of the Eucommia ulmoides group,)the Sargentodoxa group and the Phellinus igniarius group significantly decreased(P<0.05).Eucommia ulmoides and Caulis spatholobi are more effective in reducing pathological damage of toes in mice.Conclusion:The water extract of Caulis spatholobi has obvious anti-gout effect,and the most significant effect on analgesia and swelling.Eucommia ulmoides and Sargentodoxa cuneata have more obvious anti-inflammatory effects and have certain analgesic and swelling effects.Phellinus Phellinus igniarius has obvious inhibitory effect on IL-1β,but it does not show good anti-gout potential.展开更多
Objective:To investigate the neuroprotective effect of Phyllanthus urinaria on ischemic stroke and its primary mechanism.Methods:Adult SD rats were selected as the research object,and the right middle cerebral artery ...Objective:To investigate the neuroprotective effect of Phyllanthus urinaria on ischemic stroke and its primary mechanism.Methods:Adult SD rats were selected as the research object,and the right middle cerebral artery infarction rat model was established by the modified suture method(MCAO).Observe the neurological deficit score at 24h,48h and 72h after the model is successfully prepared,then TTC staining method to detect the area of cerebral infarction,the content of superoxide dismutase(SOD),nitric oxide(NO)and endothelial NOS(eNOS)in brain tissue;Immunofluorescence method was used to detect the expression of Caspase-3 positive cells in brain tissue;Western blot method was used to detect the expression of PI3K and AKT protein in brain tissue.72 experimental animals were randomly divided into 4 groups,sham operation group,model group(MCAO),extract of Phyllanthus urinaria low dose group(PuL5 g/kg),high dose group(PuL10 g/kg),and make MCAO model after 7 days of continuous administration,and continue to infuse the medicine once/d until the material is obtained.Results:No neurological deficits in the sham operation group,The 24h,48h and 72h of the modelling showed that the neurological impairment of the two doses of extract of Phyllanthus urinaria and the MCAO group was more severe than that of the sham operation group(P<0.01),however,with the prolongation of the modeling time,the neurological function scores of the two doses of extract of Phyllanthus urinaria were lower than those of the MCAO group,the most significant at 72h(P<0.01);The infarct size of the rats in the two dose groups of extract of Phyllanthus urinaria was lower than that of the MCAO group(P<0.01),and there was no dose dependence between the two groups,the content of SOD in the MCAO group was reduced,and the content of NO and eNOS was increased than the sham operation group(P<0.05),Compared with the MCAO group,the two administration groups significantly increased the content of SOD,decreased the content of NO and eNOS(P<0.05);Although the expression of Caspase-3 positive cells in the two administration groups was higher than that in the sham operation group,it was significantly lower than that in the MCAO group(Plow<0.05,Phigh<0.01);The expression of PI3K and AKT protein in the brain tissue of the MCAO group was significantly lower than that of the sham operation group(P<0.05),but the expression of PI3K and AKT protein in brain tissue of extract of Phyllanthus urinaria in high and low dose groups was significantly higher than that in MCAO group(P<0.05).Conclusion:Extract of Phyllanthus urinaria can improve neurological damage and cerebral infarction area in rats,and reduce the expression of Caspase-3 positive cells in MCAO rats,and then increase the expression levels of PI3K and AKT proteins to protect ischemic brain injury.展开更多
Objective:To investigate the effect and anti-inflammatory mechanism the effect of Flemingia Roxb.exAit extract on acute gouty arthritis.Methods:Sixty SD rats were randomly divided into 6 groups by weight,control and m...Objective:To investigate the effect and anti-inflammatory mechanism the effect of Flemingia Roxb.exAit extract on acute gouty arthritis.Methods:Sixty SD rats were randomly divided into 6 groups by weight,control and model group,(100、150、200 mg/kg)of Flemingia Roxb.exAit extract group and colchicine group.The rats of each group were given by intragastric for 7 days of continuous administration,meanwhile,there were given 0.9%NaCl solution instead as control and model group.The acute gouty arthritis model was constructed through right ankle joint cavity injection of sodium urate crystal solutionon day 5 after 1 h,but rats in the negative control group were injected with 0.9%NaCl solution into the articular cavity of the right foot.The general condition of rats and the degree of joint swelling,and the gait was observed after constructed model.To detected the joint fluid IL-1β、TNF-α、IL-6 indicators,and the expression of NLRP3,Caspase-1 protein in ankle joint tissue for 2 hours after the last administration.Results:Groups of Flemingia Roxb.exAit extract could markedly reduce the joint swelling on acute gouty arthritis rat model,improve gait in a dose-dependent manner,high dose group of Flemingia Roxb.exAit extract did particularly well(Pjoint swelling<0.01;Pgait<0.05),close to the therapeutic effect colchicine group.Further mechanism studies show that the joint fluid IL-1β、TNF-α、IL-6 level was reduced by Flemingia Roxb.exAit extract group,the expressions of NLRP3、Caspase-1 protein of Flemingia Roxb.exAit extract group was decreased observbly than model group,but the most obvious is that high dose group as well as colchicine group(PNALP3<0.01;PCaspase-1<0.01).Conclusion:Flemingia Roxb.exAit extract has the effect of treating acute gouty arthritis and its mechanism may be related toinhibiting NLRP3 inflammasome activation and inhibiting inflammatory response.展开更多
Objective:To investigate the protective effect of Tadehaginoside on vascular endothelial cell injury induced by reactive nitrogen.Methods:MTT colorimetry was used to detect the effect of Tadehaginoside on the survival...Objective:To investigate the protective effect of Tadehaginoside on vascular endothelial cell injury induced by reactive nitrogen.Methods:MTT colorimetry was used to detect the effect of Tadehaginoside on the survival rate of EA.hy 926 endothelial cells in the concentration range of 5~160μmol/L;1 h after pre-administration of Tadehaginoside,0.5 mM GSNO was given to damage endothelial cells.Detect the mitochondrial specific factors COX-1,ND-1 and inflammatory factor IL-1βof EA.hy 926 cells damaged by GSNO by Real time-PCR method gene intervention.At the same time,Western blot was used to detect the changes in Bax and Bcl-2 protein expression.The mitochondrial membrane potential kit(JC-1)was used to detect the change of Tadehaginoside on the mitochondrial membrane potential after GSNO induced EA.hy 926 cell injury.Results:The results of the MTT method showed that Tadehaginoside had no obvious cytotoxicity on EA.hy 926 cells in the range of 5~160μmol/L,and the optimal protective concentration of the drug was 40μmol/L.Western Blot method showed that BAX protein expression increased in a time-dependent manner after GSNO damaged EA.hy 926 cells over time,while Bcl-2 protein expression was the opposite.Real time-PCR results showed that Tadehaginoside can significantly up-regulate COX-1 gene(P<0.05),and can significantly inhibit GSNO induced ND-1(P<0.05)and IL-1βgene up-regulation(P<0.01).At the same time,the results of JC-1 showed that Tadehaginoside could significantly protect the mitochondrial membrane potential from GSNO damage.Conclusion:The GSNO damage model may induce the increase of Bax and other pro-apoptotic proteins through mitochondrial DNA damage and reduce the expression of anti-apoptotic factor Bcl-2.Tadehaginoside has a certain protective effect on endothelial cell mitochondrial damage induced by reactive nitrogen,and its mechanism is related to inhibiting the expression of ND-1 and IL-1βgenes and upregulating the expression of COX-1 genes.展开更多
Objective:To investigate the effect ofPhyllanthusurinaria L extract on hyperuricemia mice.Methods:We were randomly divided into 6 groups by weight,control and model group,(15、30、45 mg/kg)ofPhyllanthusurinaria L extr...Objective:To investigate the effect ofPhyllanthusurinaria L extract on hyperuricemia mice.Methods:We were randomly divided into 6 groups by weight,control and model group,(15、30、45 mg/kg)ofPhyllanthusurinaria L extract group and allopurinol group.There were given oteracil potassium 300 mg/kg intragastrically to induce hyperuricemiamodel without control group,Phyllanthusurinaria L extract were given by intragastric administration,there were given saline solution instead as control and model group.Uric acid,blood urea ni-trogen and creatinine was detected in serum in n hyperuricemia mice for intragastric administration eight days.the contents and activity of xanthine oxidasein liver tissue were measured in hyperuricemia mice.The pathological changes of renal tissues were observed with HE staining in groups of mice.Western blot was surveied the expression of URAT1 protein in mice kidney tissues.Results:Groups ofPhyllanthusurinaria L extract could markedly reduce the uric acid,blood urea ni-trogen and creatinine level in serum on hyperuricemia mice,at the same time it reduce the contents and activity of liver in hyperuricemia mice,high dose group of Phyllanthusurinaria L extract did particularly well,close to allopurinol group,it could improve the pathologicalchanges in kidney tissueon hyperuricemia mice better than allopurinolgroup.the expressions of URAT1 protein of the model group was increased observbly than control group(P<0.05),but high dose group ofPhyllanthusurinaria L extract group wasdecreased more than model group,same level as control group(P<0.05).Conclusion:Phyllanthusurinaria L extractcould effectively were reduced the level of Uric acid in hyperuricemia mice,there were possibly reducing XOD activity,meanwhile,there were restrained the protein expression of URAT1 and decreased morphological changes in hyperuricemia mice.展开更多
基金supported by grants of Hainan provincial project of modernization in traditional Chinese medicine(ZY201426)the Key Science and Technology Program of Hainan Province(ZDXM2014070)the National Natural Science Foundation of China(81460550)
文摘Objective:To determine the chemical structure of the new compound and investigate the protective effects of Tinosporaic acid A and B towards in-vitro neuro.Methods:The structures of two new compounds were established by analyzing its 1D and 2D NMR spectra as well as HRESIMS.Their neuroprotective effects with respect to the antioxidant properties were evaluated by radical scavenging tests and hydrogen peroxide-injured oxidative stress model in PC12 cell lines.Cell morphology of treated PC12 cells was observed by phase contrast microscopy.In-vitro MTT assay,lactate dehydrogenase activity assay and oxidative stress markers(intracellular ROS production,MDA level,and caspase-3 activity) were used to evaluate the protective effects against hydrogen peroxide induced cytotoxicity in PC12 cells.Results:The two new compounds,named Tinosporaic acid A and B,were isolated and identified from the stem bark of Tinospora hainanensis.Cell viability studies identified a representative concentration for each extract that was subsequently used to measure oxidative stress markers.Both extracts were able to reverse the oxidative damage caused by hydrogen peroxide,thus promoting PC12 cells survival.The concentration of Tinosporaic acid A and B were 86.34 μg/mL and 22.06 μg/mL respectively,which is neuroprotective for EC_(50).The results indicated that both of them significantly attenuated hydrogen peroxide-induced neurotoxicity.Conclusion:The two new compounds isolated from ethanol extracts of Tinospora hainanensis are the promising natural ones with neuroprotective activity and needed for further research.
基金National Natural Science Foundation of China(No.81960663)2016 Student Innovation Training Program of Hainan Medical University(No.HYCX2016055)+1 种基金2017 College Student Innovation and Entrepreneurship Training Program of Hainan Province(No.2017098)2018 Research Start Funding of Hainan Medical University。
文摘Objective:To explore the anti-gout effects of four traditional Chinese medicine extracts of Eucommia,Sargentodoxa cuneata,Caulis spatholobi and Phellinus igniarius.Methods:Monosodium Urate(MSU)was used to establish a mouse gouty arritis model.With coloricine as the positive control group,the effects of four traditional Chinese medicine extracts of Ecuador,Sargentodoxa,Caulis spatholobi and Phellinus igniarius on rice were studied;The hot plate method was used to observe the analytical effects of the four groups of traditional Chinese medicine extracts;the enzyme-linked immunoassay(ELISA)was used to detect residual interference-1 beta(IL-1 beta);the four traditional Chinese medicine extracts were simultaneously determined on the white blood cells,and monocytes in the blood of mice as well as the influence of neutrophils and histopathological changes of toes.Results:Compared with the normal group,the joint diameter ratio,blood leukocytes,monocytes,neutrophils,IL-1βat different time points in the model group were significantly increased(P<0.01),and the pain percentage was significantly reduced(P<0.05).Compared with the model group,the joint diameter ratio of the colchicine group,the Eucommia group,the Sargentodoxa group,the Caulis spatholobi group and the Phellinus sibiricum group was significantly reduced(P<0.05),and the pain percentage was significantly increased(P(0.05).The four groups of traditional Chinese medicine extracts significantly decreased the white blood cells,monocytes,and neutrophils in the blood of mice(P<0.05),and the IL-1βexpression levels of the Eucommia ulmoides group,)the Sargentodoxa group and the Phellinus igniarius group significantly decreased(P<0.05).Eucommia ulmoides and Caulis spatholobi are more effective in reducing pathological damage of toes in mice.Conclusion:The water extract of Caulis spatholobi has obvious anti-gout effect,and the most significant effect on analgesia and swelling.Eucommia ulmoides and Sargentodoxa cuneata have more obvious anti-inflammatory effects and have certain analgesic and swelling effects.Phellinus Phellinus igniarius has obvious inhibitory effect on IL-1β,but it does not show good anti-gout potential.
基金Natural Science Foundation of Hainan Province(No.817132)。
文摘Objective:To investigate the neuroprotective effect of Phyllanthus urinaria on ischemic stroke and its primary mechanism.Methods:Adult SD rats were selected as the research object,and the right middle cerebral artery infarction rat model was established by the modified suture method(MCAO).Observe the neurological deficit score at 24h,48h and 72h after the model is successfully prepared,then TTC staining method to detect the area of cerebral infarction,the content of superoxide dismutase(SOD),nitric oxide(NO)and endothelial NOS(eNOS)in brain tissue;Immunofluorescence method was used to detect the expression of Caspase-3 positive cells in brain tissue;Western blot method was used to detect the expression of PI3K and AKT protein in brain tissue.72 experimental animals were randomly divided into 4 groups,sham operation group,model group(MCAO),extract of Phyllanthus urinaria low dose group(PuL5 g/kg),high dose group(PuL10 g/kg),and make MCAO model after 7 days of continuous administration,and continue to infuse the medicine once/d until the material is obtained.Results:No neurological deficits in the sham operation group,The 24h,48h and 72h of the modelling showed that the neurological impairment of the two doses of extract of Phyllanthus urinaria and the MCAO group was more severe than that of the sham operation group(P<0.01),however,with the prolongation of the modeling time,the neurological function scores of the two doses of extract of Phyllanthus urinaria were lower than those of the MCAO group,the most significant at 72h(P<0.01);The infarct size of the rats in the two dose groups of extract of Phyllanthus urinaria was lower than that of the MCAO group(P<0.01),and there was no dose dependence between the two groups,the content of SOD in the MCAO group was reduced,and the content of NO and eNOS was increased than the sham operation group(P<0.05),Compared with the MCAO group,the two administration groups significantly increased the content of SOD,decreased the content of NO and eNOS(P<0.05);Although the expression of Caspase-3 positive cells in the two administration groups was higher than that in the sham operation group,it was significantly lower than that in the MCAO group(Plow<0.05,Phigh<0.01);The expression of PI3K and AKT protein in the brain tissue of the MCAO group was significantly lower than that of the sham operation group(P<0.05),but the expression of PI3K and AKT protein in brain tissue of extract of Phyllanthus urinaria in high and low dose groups was significantly higher than that in MCAO group(P<0.05).Conclusion:Extract of Phyllanthus urinaria can improve neurological damage and cerebral infarction area in rats,and reduce the expression of Caspase-3 positive cells in MCAO rats,and then increase the expression levels of PI3K and AKT proteins to protect ischemic brain injury.
基金2016 Hainan Medical College Students Innovation and Entrepreneurship Training Program(No.HYCX2016055)2017 Hainan College Students Innovation and Entrepreneurship Training Program Project(No.2017098)。
文摘Objective:To investigate the effect and anti-inflammatory mechanism the effect of Flemingia Roxb.exAit extract on acute gouty arthritis.Methods:Sixty SD rats were randomly divided into 6 groups by weight,control and model group,(100、150、200 mg/kg)of Flemingia Roxb.exAit extract group and colchicine group.The rats of each group were given by intragastric for 7 days of continuous administration,meanwhile,there were given 0.9%NaCl solution instead as control and model group.The acute gouty arthritis model was constructed through right ankle joint cavity injection of sodium urate crystal solutionon day 5 after 1 h,but rats in the negative control group were injected with 0.9%NaCl solution into the articular cavity of the right foot.The general condition of rats and the degree of joint swelling,and the gait was observed after constructed model.To detected the joint fluid IL-1β、TNF-α、IL-6 indicators,and the expression of NLRP3,Caspase-1 protein in ankle joint tissue for 2 hours after the last administration.Results:Groups of Flemingia Roxb.exAit extract could markedly reduce the joint swelling on acute gouty arthritis rat model,improve gait in a dose-dependent manner,high dose group of Flemingia Roxb.exAit extract did particularly well(Pjoint swelling<0.01;Pgait<0.05),close to the therapeutic effect colchicine group.Further mechanism studies show that the joint fluid IL-1β、TNF-α、IL-6 level was reduced by Flemingia Roxb.exAit extract group,the expressions of NLRP3、Caspase-1 protein of Flemingia Roxb.exAit extract group was decreased observbly than model group,but the most obvious is that high dose group as well as colchicine group(PNALP3<0.01;PCaspase-1<0.01).Conclusion:Flemingia Roxb.exAit extract has the effect of treating acute gouty arthritis and its mechanism may be related toinhibiting NLRP3 inflammasome activation and inhibiting inflammatory response.
基金National Natural Science Fund(No.81960663)College Students Innovation Training Program for Hainan Medical College in 2019(No.X201911810036)。
文摘Objective:To investigate the protective effect of Tadehaginoside on vascular endothelial cell injury induced by reactive nitrogen.Methods:MTT colorimetry was used to detect the effect of Tadehaginoside on the survival rate of EA.hy 926 endothelial cells in the concentration range of 5~160μmol/L;1 h after pre-administration of Tadehaginoside,0.5 mM GSNO was given to damage endothelial cells.Detect the mitochondrial specific factors COX-1,ND-1 and inflammatory factor IL-1βof EA.hy 926 cells damaged by GSNO by Real time-PCR method gene intervention.At the same time,Western blot was used to detect the changes in Bax and Bcl-2 protein expression.The mitochondrial membrane potential kit(JC-1)was used to detect the change of Tadehaginoside on the mitochondrial membrane potential after GSNO induced EA.hy 926 cell injury.Results:The results of the MTT method showed that Tadehaginoside had no obvious cytotoxicity on EA.hy 926 cells in the range of 5~160μmol/L,and the optimal protective concentration of the drug was 40μmol/L.Western Blot method showed that BAX protein expression increased in a time-dependent manner after GSNO damaged EA.hy 926 cells over time,while Bcl-2 protein expression was the opposite.Real time-PCR results showed that Tadehaginoside can significantly up-regulate COX-1 gene(P<0.05),and can significantly inhibit GSNO induced ND-1(P<0.05)and IL-1βgene up-regulation(P<0.01).At the same time,the results of JC-1 showed that Tadehaginoside could significantly protect the mitochondrial membrane potential from GSNO damage.Conclusion:The GSNO damage model may induce the increase of Bax and other pro-apoptotic proteins through mitochondrial DNA damage and reduce the expression of anti-apoptotic factor Bcl-2.Tadehaginoside has a certain protective effect on endothelial cell mitochondrial damage induced by reactive nitrogen,and its mechanism is related to inhibiting the expression of ND-1 and IL-1βgenes and upregulating the expression of COX-1 genes.
基金2016 Hainan medical university students’innovation and entrepreneurship training program(No.HYCX2016042)2017 Hainan university students’innovation and entrepreneurship training program(No.2017085)
文摘Objective:To investigate the effect ofPhyllanthusurinaria L extract on hyperuricemia mice.Methods:We were randomly divided into 6 groups by weight,control and model group,(15、30、45 mg/kg)ofPhyllanthusurinaria L extract group and allopurinol group.There were given oteracil potassium 300 mg/kg intragastrically to induce hyperuricemiamodel without control group,Phyllanthusurinaria L extract were given by intragastric administration,there were given saline solution instead as control and model group.Uric acid,blood urea ni-trogen and creatinine was detected in serum in n hyperuricemia mice for intragastric administration eight days.the contents and activity of xanthine oxidasein liver tissue were measured in hyperuricemia mice.The pathological changes of renal tissues were observed with HE staining in groups of mice.Western blot was surveied the expression of URAT1 protein in mice kidney tissues.Results:Groups ofPhyllanthusurinaria L extract could markedly reduce the uric acid,blood urea ni-trogen and creatinine level in serum on hyperuricemia mice,at the same time it reduce the contents and activity of liver in hyperuricemia mice,high dose group of Phyllanthusurinaria L extract did particularly well,close to allopurinol group,it could improve the pathologicalchanges in kidney tissueon hyperuricemia mice better than allopurinolgroup.the expressions of URAT1 protein of the model group was increased observbly than control group(P<0.05),but high dose group ofPhyllanthusurinaria L extract group wasdecreased more than model group,same level as control group(P<0.05).Conclusion:Phyllanthusurinaria L extractcould effectively were reduced the level of Uric acid in hyperuricemia mice,there were possibly reducing XOD activity,meanwhile,there were restrained the protein expression of URAT1 and decreased morphological changes in hyperuricemia mice.
