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
The study on the element transference characteristics and element speciation in the extract of Cyclocarya paliurus(Batal.) Iljinskaja leaves was performed by FIA-ICP-AES on the basis of C-18 bonded silica gel column s...The study on the element transference characteristics and element speciation in the extract of Cyclocarya paliurus(Batal.) Iljinskaja leaves was performed by FIA-ICP-AES on the basis of C-18 bonded silica gel column separation and microwave digestion/extraction for sample treatment. Higher transfer ratio could be obtained by artificial gastric acid or with microwave extraction method. The contents of inorganic species and organic species of the four elements in the microwave extract at the temperature of 100℃ are as follows. Zn: (4. 46±0.13) μg/g and (8. 16±0. 15) μg/g; Mn: (35. 7±1.1) μg/g and (91. 2± 3.2μg/g); Mg: (156±9)μg/g and (513±11)μg/g; Fe: (3. 28±0.19)μg/g and (8. 43±1.15) μg/g, respectively.展开更多
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.
文摘The study on the element transference characteristics and element speciation in the extract of Cyclocarya paliurus(Batal.) Iljinskaja leaves was performed by FIA-ICP-AES on the basis of C-18 bonded silica gel column separation and microwave digestion/extraction for sample treatment. Higher transfer ratio could be obtained by artificial gastric acid or with microwave extraction method. The contents of inorganic species and organic species of the four elements in the microwave extract at the temperature of 100℃ are as follows. Zn: (4. 46±0.13) μg/g and (8. 16±0. 15) μg/g; Mn: (35. 7±1.1) μg/g and (91. 2± 3.2μg/g); Mg: (156±9)μg/g and (513±11)μg/g; Fe: (3. 28±0.19)μg/g and (8. 43±1.15) μg/g, respectively.
