The biological activity of plant polysaccharides can be enhanced by sulfated modification.In this study,the immunomodulatory effect of sulfated Cyclocarya paliurus polysaccharides(SCP3)on macrophages RAW264.7 and its ...The biological activity of plant polysaccharides can be enhanced by sulfated modification.In this study,the immunomodulatory effect of sulfated Cyclocarya paliurus polysaccharides(SCP3)on macrophages RAW264.7 and its potential molecular mechanism were investigated.Results showed that SCP3 at 25-100μg/m L increased viability and improved phagocytosis of RAW264.7 cells.Meanwhile,SCP3 could activate mitogen-activated protein kinase(MAPK)and nuclear factor kappa B(NF-κB)signaling pathways,which increased the phosphorylation of Erk1/2,JNK,p38 and NF-κB p65,promoting secretion of cytokines tumor necrosis factorα(TNF-α),interleukin 6(IL-6)and nitric oxide(NO)as well as the production of reactive oxygen species(ROS).In addition,Toll-like receptor 4(TLR4)receptor inhibitors were able to block the production of NO and TNF-αby SCP3-stimulated macrophages.Based on Western blot analysis and validation using specific inhibitors against MAPK and NF-κB signaling pathways,the results demonstrated that SCP3 induced macrophages activation and enhanced TNF-αand NO production via TLR4-mediated MAPK and NF-κB pathways.In summary,SCP3 has significant immunomodulatory potential.The underlying molecular mechanism was that SCP3 activates macrophages via TLR4 receptors to promote ROS production,which in turn activates the downstream MAPK/NF-κB signaling pathway and then increases the secretion levels of cytokines and NO.展开更多
In this study,the antitumor properties and the possible molecular mechanisms of Cyclocarya paliurus polysaccharide(CP)and its phosphorylated derivative(P-CP)on CT-26 mouse colon carcinoma cells were investigated.Resul...In this study,the antitumor properties and the possible molecular mechanisms of Cyclocarya paliurus polysaccharide(CP)and its phosphorylated derivative(P-CP)on CT-26 mouse colon carcinoma cells were investigated.Results found that CP had high inhibition ratio against CT-26 cells.The flow cytometry results found that CP treatment could cause the intracellular acidification,arrest the cell cycle in the S phase and increase reactive oxygen species generation.Additionally,CP treatment triggered mitochondrial membrane potential depolarization and Ca^(2+)overloading,and broke down the balance of antioxidant system,Na^(+)/K^(+)-ATPase and Ca^(2+)-ATPase.Further analysis found CP induced cell apoptosis through improving the activities of caspase-3 and caspase-9,and increasing the level of cytochrome C.Furthermore,the comparative study of antitumor effect on CT-26 cells displayed that the phosphorylation enhanced antitumor activities of polysaccharides.These results suggest CP is a potential natural therapeutic agent for colon cancer and phosphorylation represents an effective method of enhancing the antitumor activity of CP.展开更多
The ultrastructural distribution and active location of ATPase and the ultrastructural variations were investigated in mesophyll cells of Cyclocarya paliurus seedlings after iso-osmotic salt/water treatments in combin...The ultrastructural distribution and active location of ATPase and the ultrastructural variations were investigated in mesophyll cells of Cyclocarya paliurus seedlings after iso-osmotic salt/water treatments in combination with calcium regulation. C. paliurus seed- lings were treated with five groups (control, 85 mM NaCl, 85 mM NaCl+ 12 mM Ca(NO3)2, PEG iso-osmotic to 85 mM NaCl and PEG iso-osmotic to 85 mM NaCl+12 mM Ca(NO))2) in a hydroponic system in a phytotron. Results show that under normal growth conditions the ATPase activity was low and the enzyme was primarily located on the nucleus. After 12 days of iso-osmotic salt/water treatments, ATPase activity on the tonoptast increased. Osmiophilic globules for iso-osmotic water treatment were greater than that for iso-osmotie salt treatments. The ATPase activity increased and was mostly transferred onto the nucleus for calcium regulation treatment under iso- osmotic salt/water stresses, and the osmiophilic globules significantly decreased under iso-osmotic water stress with calcium regulation. The ATPase located on the nucleus indicated that the degree of salt/drought damage that seedlings suffered was slighter, while the amount of the enzyme located on the tonoplast showed that the degree of salt/drought damage there was more serious. After 4 and 20 days of iso- osmotic treatments, the injury suffered by the leaf ultrastructures of C. paliurus seedlings for iso-osmotic treatment with calcium regula- tion was lower than those without calcium regulation, especially for the iso-osmotic water treatments. Preliminary analysis suggests that the iniury suffered by C. paliurus seedlings was lower for iso-osmotic salt treatments than for iso-osmotic water treatments, while the effect of calcium regulation under iso-osmotic water stress was greater than that of the iso-osmotic salt stress.展开更多
Cyclocarya paliurus, an economically valuable tree species, has traditionally been used as a nutraceutical food or medicine in China. However, limited information is available on its genotype selection and cultivation...Cyclocarya paliurus, an economically valuable tree species, has traditionally been used as a nutraceutical food or medicine in China. However, limited information is available on its genotype selection and cultivation under a wide range of environmental conditions for growth and targeted health-promoting substances. We studied the effects of genotype and environment, and their interaction on leaf triterpenoid content and tree growth for 12 genotypes of C. paliurus grown at four sites. We quantified the correlation between leaf triterpenoid accumulation and tree growth. The contents of cyclocaric acid B, cyclocarioside I, and arjunolic acid ranged from 0.06 to 3.89, 0 to 3.71, and 0.65 to 8.86 mg g^-1, respectively. Three individual triterpenoids were primarily influenced by genotype (variation ranged from 53.7 to 68.0%), while environment accounted for most of the variation in total triterpenoid content and tree growth (71.3–89.5%). Most tested environmental parameters were significantly correlated to total triterpenoid content, but not to the contents of the individual triterpenoids measured. Growth in tree height and diameter at breast height were significantly negatively correlated with total triterpenoid content but were non-significantly correlated with individual triterpenoid contents. We conclude that genotypic selection, manipulation of environmental conditions, and implementation of appropriate silvicultural operations would be important strategies for increasing the accumulation of health-promoting phytochemicals.展开更多
Cyclocarya paliurus is widely distributed in subtropical areas of China.Secondary metabolites in the leaves,including flavonoids and triterpenoids,provide protection against diseases such as hyperlipidemia,diabetes,hy...Cyclocarya paliurus is widely distributed in subtropical areas of China.Secondary metabolites in the leaves,including flavonoids and triterpenoids,provide protection against diseases such as hyperlipidemia,diabetes,hypertension,and hypoimmunity.In this study,343 C.paliurus leaves were collected from 32 distinct populations covering most of the main distribution,to deduce the response of flavonoids and triterpenoids to seven environmental factors.Principal component analysis was performed using seven flavonoid and six triterpenoid compounds.The results show that altitude,latitude,and longitude significantly contributed to variations in total flavonoid and triterpenoid contents.In addition,the interaction of these metabolites was extensively correlated with environment changes,and therefore should be considered during selection and cultivation.展开更多
OBJECTIVE To explore the protective effects of total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja on STZ-stimulated INS-1 cells through regulating of autophagy and apoptosis.METHODS INS-1cells wer...OBJECTIVE To explore the protective effects of total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja on STZ-stimulated INS-1 cells through regulating of autophagy and apoptosis.METHODS INS-1cells were cultured in media containing 3n M STZ and different doses of total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja.The proliferation of cells was examined by MTT assay,ROS content were detected by fluorescence enzyme label.The levels of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),hydrogen peroxidase(CAT),malondialdehyde(MDA)were also measured by colorimetry.The activities of caspase-3,9 were also observed by Caspase colorimetric assay kit in each group.The expressions of Bcl-2 m RNA and Bax m RNA were detected by Real time PCR,protein expression of LC3-Ⅱand PARP were detected by Western blotting.RESULTS Compared with the control group,total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja could promote the proliferation of INS-1 cells no matter with STZ or not when its concentration lower than 25μg·m L-1;but when its concentration higher than 100μg·m L-(1use individually)or 50μg·m L-1(combined use),total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja might significantly inhibite the growth of the cells whether STZ existed or not.Total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja(6.25,12.5,25μg·m L-1)might inhibit INS-1cell apoptosis,decrease intra-cellular ROS contents,improve the s upernatant liquid SOD,GSH-PX,CAT activities,decrease MDA level,promote INS-1 cell secreting insulin,decrease the protein expressions of autophagy protein LC3Ⅱand apoptosis regulating protein cleaved PARP protein,up-regulate the anti-apoptotic protein Bcl-2 m RNA expression,down-regulate the pro-apoptotic protein Bax m RNA expression and Bcl-2and Bax,reduce the activity of caspase-9 and caspase-3(P<0.05 or P<0.01).CONCLUSION Total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja has good protective effect on STZ-stimulated INS-1cells.It can inhibit STZ injured INS-1 cells to overproduce ROS production,enhance endogenous antioxidant enzymes(GSH-Px,SOD,CAT activities),reduce the expression of autophagy protein LC3Ⅱand apoptosisregulating protein cleaved PARP,up-reglute the antiapoptosis protein Bcl-2 expression,down-regulate the pro-apoptotic protein Bax expression,decrease the caspase-9 and caspase-3 activities,and improve the INS-1cell survival rate,and then play a protective effect on damaged INS-1 cells.展开更多
The hormone combinations and dark culture of the callus induction for Cyclocarya paliurus (Batal.) Iljinskaja leaves were optimized by an orthogonal experiment. The effects of hormones on callus proliferation were a...The hormone combinations and dark culture of the callus induction for Cyclocarya paliurus (Batal.) Iljinskaja leaves were optimized by an orthogonal experiment. The effects of hormones on callus proliferation were also studied. The results show that the optimum medium for callus induction of C. paliurus leaves was MS + 1.0 mg.L-1 6-BA + 0.5 mg.L-1 NAA + 1.0 mg·L-1 2,4-D. The combination of 6-BA and NAA significantly affected the induction of callus, while the effect of 2,4-D was less. The maximum rate of callus induction was 95.4%. A 10-day darkness period was beneficial for callus induction. The best proliferation medium was 1/4 DKW + 0.5 mg·L-1 6-BA + 1.0 mg·L-1 2,4-D.展开更多
[Objectives]This study was conducted to discuss the mechanism of the blood sugar-lowering effect of Cyclocarya paliurus extract.[Methods]With C.paliurus leaves as the raw material,the ethyl acetate-butyl acetate micro...[Objectives]This study was conducted to discuss the mechanism of the blood sugar-lowering effect of Cyclocarya paliurus extract.[Methods]With C.paliurus leaves as the raw material,the ethyl acetate-butyl acetate microwave extraction process of C.paliurus was studied.With the yield of C.paliurus extract as an evaluation index,single-factor experiments were carried out on such 6 factors as the dry C.paliurus leaf powder size,solid-to-liquid ratio,extraction time,microwave frequency,microwave power,and microwave extraction time,and the extraction and purification process was further optimized by response surface analysis.The normal mice and the hyperglycemic mice modeled by alloxan were injected with C.paliurus extract to study the blood sugar-lowering effects of different groups of C.paliurus extract.[Results]The optimal process combination for the extraction of C.paliurus leaves was obtained:dry C.paliurus powder size 90-100 mesh,solid-to-liquid ratio 1∶20(g/ml),extraction time 192 min,microwave frequency 2500 MHz,microwave power 490 W,and extraction time 248 s.Under these conditions,the yield of C.paliurus extract reached the maximum,95.10%,reaching 99.83%of the predicted value.The blood sugar-lowering test on mice showed that the C.paliurus extract had a good effect on lowering blood sugar,indicating that C.paliurus extract can improve the free radical scavenging capacity to a certain extent.[Conclusions]This study provides certain reference for the in-depth study of the physiological effects of C.paliurus extract and the comprehensive development and utilization of C.paliurus extract.展开更多
基金the financial supports by the National Natural Science Foundation of China(82060594)the Natural Science Foundation of Jiangxi Province,China(20202BAB205006)。
文摘The biological activity of plant polysaccharides can be enhanced by sulfated modification.In this study,the immunomodulatory effect of sulfated Cyclocarya paliurus polysaccharides(SCP3)on macrophages RAW264.7 and its potential molecular mechanism were investigated.Results showed that SCP3 at 25-100μg/m L increased viability and improved phagocytosis of RAW264.7 cells.Meanwhile,SCP3 could activate mitogen-activated protein kinase(MAPK)and nuclear factor kappa B(NF-κB)signaling pathways,which increased the phosphorylation of Erk1/2,JNK,p38 and NF-κB p65,promoting secretion of cytokines tumor necrosis factorα(TNF-α),interleukin 6(IL-6)and nitric oxide(NO)as well as the production of reactive oxygen species(ROS).In addition,Toll-like receptor 4(TLR4)receptor inhibitors were able to block the production of NO and TNF-αby SCP3-stimulated macrophages.Based on Western blot analysis and validation using specific inhibitors against MAPK and NF-κB signaling pathways,the results demonstrated that SCP3 induced macrophages activation and enhanced TNF-αand NO production via TLR4-mediated MAPK and NF-κB pathways.In summary,SCP3 has significant immunomodulatory potential.The underlying molecular mechanism was that SCP3 activates macrophages via TLR4 receptors to promote ROS production,which in turn activates the downstream MAPK/NF-κB signaling pathway and then increases the secretion levels of cytokines and NO.
基金supported by the National Natural Science Foundation of China (82060594)the Natural Science Foundation of Jiangxi province,China (20202BAB205006)The National Youth Talent Support Program of China。
文摘In this study,the antitumor properties and the possible molecular mechanisms of Cyclocarya paliurus polysaccharide(CP)and its phosphorylated derivative(P-CP)on CT-26 mouse colon carcinoma cells were investigated.Results found that CP had high inhibition ratio against CT-26 cells.The flow cytometry results found that CP treatment could cause the intracellular acidification,arrest the cell cycle in the S phase and increase reactive oxygen species generation.Additionally,CP treatment triggered mitochondrial membrane potential depolarization and Ca^(2+)overloading,and broke down the balance of antioxidant system,Na^(+)/K^(+)-ATPase and Ca^(2+)-ATPase.Further analysis found CP induced cell apoptosis through improving the activities of caspase-3 and caspase-9,and increasing the level of cytochrome C.Furthermore,the comparative study of antitumor effect on CT-26 cells displayed that the phosphorylation enhanced antitumor activities of polysaccharides.These results suggest CP is a potential natural therapeutic agent for colon cancer and phosphorylation represents an effective method of enhancing the antitumor activity of CP.
基金supported by the National Natural Science Foundation of China (Project No: 30371156)Research Foundation of Jiangsu Province (Project No: BG2006314)
文摘The ultrastructural distribution and active location of ATPase and the ultrastructural variations were investigated in mesophyll cells of Cyclocarya paliurus seedlings after iso-osmotic salt/water treatments in combination with calcium regulation. C. paliurus seed- lings were treated with five groups (control, 85 mM NaCl, 85 mM NaCl+ 12 mM Ca(NO3)2, PEG iso-osmotic to 85 mM NaCl and PEG iso-osmotic to 85 mM NaCl+12 mM Ca(NO))2) in a hydroponic system in a phytotron. Results show that under normal growth conditions the ATPase activity was low and the enzyme was primarily located on the nucleus. After 12 days of iso-osmotic salt/water treatments, ATPase activity on the tonoptast increased. Osmiophilic globules for iso-osmotic water treatment were greater than that for iso-osmotie salt treatments. The ATPase activity increased and was mostly transferred onto the nucleus for calcium regulation treatment under iso- osmotic salt/water stresses, and the osmiophilic globules significantly decreased under iso-osmotic water stress with calcium regulation. The ATPase located on the nucleus indicated that the degree of salt/drought damage that seedlings suffered was slighter, while the amount of the enzyme located on the tonoplast showed that the degree of salt/drought damage there was more serious. After 4 and 20 days of iso- osmotic treatments, the injury suffered by the leaf ultrastructures of C. paliurus seedlings for iso-osmotic treatment with calcium regula- tion was lower than those without calcium regulation, especially for the iso-osmotic water treatments. Preliminary analysis suggests that the iniury suffered by C. paliurus seedlings was lower for iso-osmotic salt treatments than for iso-osmotic water treatments, while the effect of calcium regulation under iso-osmotic water stress was greater than that of the iso-osmotic salt stress.
基金supported by the National Natural Science Foundation of China(Grant No.31470637)the Collaborative Innovation Plan of Jiangsu Higher Educationthe Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)
文摘Cyclocarya paliurus, an economically valuable tree species, has traditionally been used as a nutraceutical food or medicine in China. However, limited information is available on its genotype selection and cultivation under a wide range of environmental conditions for growth and targeted health-promoting substances. We studied the effects of genotype and environment, and their interaction on leaf triterpenoid content and tree growth for 12 genotypes of C. paliurus grown at four sites. We quantified the correlation between leaf triterpenoid accumulation and tree growth. The contents of cyclocaric acid B, cyclocarioside I, and arjunolic acid ranged from 0.06 to 3.89, 0 to 3.71, and 0.65 to 8.86 mg g^-1, respectively. Three individual triterpenoids were primarily influenced by genotype (variation ranged from 53.7 to 68.0%), while environment accounted for most of the variation in total triterpenoid content and tree growth (71.3–89.5%). Most tested environmental parameters were significantly correlated to total triterpenoid content, but not to the contents of the individual triterpenoids measured. Growth in tree height and diameter at breast height were significantly negatively correlated with total triterpenoid content but were non-significantly correlated with individual triterpenoid contents. We conclude that genotypic selection, manipulation of environmental conditions, and implementation of appropriate silvicultural operations would be important strategies for increasing the accumulation of health-promoting phytochemicals.
基金supported by the Natural Science Foundation of Jiangsu Province(Grants No.BK 20160926)the Forestry Science and Technology Promotion Project from the State Forestry Administration of China[2017(08)]the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD).
文摘Cyclocarya paliurus is widely distributed in subtropical areas of China.Secondary metabolites in the leaves,including flavonoids and triterpenoids,provide protection against diseases such as hyperlipidemia,diabetes,hypertension,and hypoimmunity.In this study,343 C.paliurus leaves were collected from 32 distinct populations covering most of the main distribution,to deduce the response of flavonoids and triterpenoids to seven environmental factors.Principal component analysis was performed using seven flavonoid and six triterpenoid compounds.The results show that altitude,latitude,and longitude significantly contributed to variations in total flavonoid and triterpenoid contents.In addition,the interaction of these metabolites was extensively correlated with environment changes,and therefore should be considered during selection and cultivation.
基金The project supported by National Natural and Science Foundation of China(81341141)Yichang Qingqianliu Biotechnology Co.,Ltd.(SDHZ201602)Science and Technology Bureau of Yichang city(A15301-25)
文摘OBJECTIVE To explore the protective effects of total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja on STZ-stimulated INS-1 cells through regulating of autophagy and apoptosis.METHODS INS-1cells were cultured in media containing 3n M STZ and different doses of total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja.The proliferation of cells was examined by MTT assay,ROS content were detected by fluorescence enzyme label.The levels of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),hydrogen peroxidase(CAT),malondialdehyde(MDA)were also measured by colorimetry.The activities of caspase-3,9 were also observed by Caspase colorimetric assay kit in each group.The expressions of Bcl-2 m RNA and Bax m RNA were detected by Real time PCR,protein expression of LC3-Ⅱand PARP were detected by Western blotting.RESULTS Compared with the control group,total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja could promote the proliferation of INS-1 cells no matter with STZ or not when its concentration lower than 25μg·m L-1;but when its concentration higher than 100μg·m L-(1use individually)or 50μg·m L-1(combined use),total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja might significantly inhibite the growth of the cells whether STZ existed or not.Total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja(6.25,12.5,25μg·m L-1)might inhibit INS-1cell apoptosis,decrease intra-cellular ROS contents,improve the s upernatant liquid SOD,GSH-PX,CAT activities,decrease MDA level,promote INS-1 cell secreting insulin,decrease the protein expressions of autophagy protein LC3Ⅱand apoptosis regulating protein cleaved PARP protein,up-regulate the anti-apoptotic protein Bcl-2 m RNA expression,down-regulate the pro-apoptotic protein Bax m RNA expression and Bcl-2and Bax,reduce the activity of caspase-9 and caspase-3(P<0.05 or P<0.01).CONCLUSION Total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja has good protective effect on STZ-stimulated INS-1cells.It can inhibit STZ injured INS-1 cells to overproduce ROS production,enhance endogenous antioxidant enzymes(GSH-Px,SOD,CAT activities),reduce the expression of autophagy protein LC3Ⅱand apoptosisregulating protein cleaved PARP,up-reglute the antiapoptosis protein Bcl-2 expression,down-regulate the pro-apoptotic protein Bax expression,decrease the caspase-9 and caspase-3 activities,and improve the INS-1cell survival rate,and then play a protective effect on damaged INS-1 cells.
文摘The hormone combinations and dark culture of the callus induction for Cyclocarya paliurus (Batal.) Iljinskaja leaves were optimized by an orthogonal experiment. The effects of hormones on callus proliferation were also studied. The results show that the optimum medium for callus induction of C. paliurus leaves was MS + 1.0 mg.L-1 6-BA + 0.5 mg.L-1 NAA + 1.0 mg·L-1 2,4-D. The combination of 6-BA and NAA significantly affected the induction of callus, while the effect of 2,4-D was less. The maximum rate of callus induction was 95.4%. A 10-day darkness period was beneficial for callus induction. The best proliferation medium was 1/4 DKW + 0.5 mg·L-1 6-BA + 1.0 mg·L-1 2,4-D.
基金High-level Innovation Team and Outstanding Scholar Project of Guangxi Colleges and Universities(2019-52)"Qihuang Project"High-level Talent Team Cultivation Project of Guangxi University of Chinese Medicine(2018002)2019 Nanning City Level Scientific Research and Technology Development Planning Project(20193115)。
文摘[Objectives]This study was conducted to discuss the mechanism of the blood sugar-lowering effect of Cyclocarya paliurus extract.[Methods]With C.paliurus leaves as the raw material,the ethyl acetate-butyl acetate microwave extraction process of C.paliurus was studied.With the yield of C.paliurus extract as an evaluation index,single-factor experiments were carried out on such 6 factors as the dry C.paliurus leaf powder size,solid-to-liquid ratio,extraction time,microwave frequency,microwave power,and microwave extraction time,and the extraction and purification process was further optimized by response surface analysis.The normal mice and the hyperglycemic mice modeled by alloxan were injected with C.paliurus extract to study the blood sugar-lowering effects of different groups of C.paliurus extract.[Results]The optimal process combination for the extraction of C.paliurus leaves was obtained:dry C.paliurus powder size 90-100 mesh,solid-to-liquid ratio 1∶20(g/ml),extraction time 192 min,microwave frequency 2500 MHz,microwave power 490 W,and extraction time 248 s.Under these conditions,the yield of C.paliurus extract reached the maximum,95.10%,reaching 99.83%of the predicted value.The blood sugar-lowering test on mice showed that the C.paliurus extract had a good effect on lowering blood sugar,indicating that C.paliurus extract can improve the free radical scavenging capacity to a certain extent.[Conclusions]This study provides certain reference for the in-depth study of the physiological effects of C.paliurus extract and the comprehensive development and utilization of C.paliurus extract.