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Objective@#To investigate the regulation of curculigoside on osteogenic differentiation of MG63 and the protective effect on osteoporosis model mice.@*Methods@#The effects of curculigoside on the survival rate of dexamethasone or H2O2 treated MG63 were detected by methyl thiazolyl tetrazolium (MTT). The specimens were divided into six groups: blank control group, blank administration group, model group (dexamethasone or H2O2 treatment group), low dose group (dexamethasone or H2O2+1.0 μmol/L curculigoside), medium dose group (dexamethasone or H2O2+2.5 μmol/L curculigoside) and high dose group (dexamethasone or H2O2+5.0 μmol/L curculigoside), the sample size of each group was 10. Western blotting was used to detect the expression of osteogenic differentiation-related proteins [type Ⅰ collagen, integrin β1, osteoblast-specific transcription factor (Osterix), osteocalcin and osteopontin] in MG63 cells after 1, 7 and 14 days incubated with 0, 1.0, 2.5 and 5.0 μmol/L of curculigoside. The sample size for each group at each time point was six. The experimental mice were divided into 4 groups: blank group, model group (dexamethasone treatment group), curculigoside low-dose group (dexamethasone+5 mg/kg curculigoside) and high-dose group (dexamethasone+45 mg/kg curculigoside), twenty each. After treatment, the tibia of the mice in each group were subjected to sacral HE staining. The number of osteoclasts was counted, and the levels of oxidative related factors in serum were determined by enzyme-linked immunosorbent assay (ELISA).@*Results@#The MTT results showed that compared with the blank control group [(100±3.7)%], the cell survival rate decreased to (44.1±5.7)% after treatment with dexamethasone, and the survival rate increased to (79.7±3.8)% after treatment with 5.0 μmol/L of curculigoside. The cell survival rate decreased to (59.1±4.7)% after H2O2 treatment, and the survival rate increased to (80.8±3.5)% after treatment with 2.5 μmol/L of curculigoside. The results of Western blotting showed that the expression of type Ⅰ collagen and integrin β1 in MG63 cells was significantly increased after 1.0, 2.5 and 5.0 μmol/L of curculigoside for 1, 7 and 14 days compared with 0 μmol/L of curculigo side for the same period. After increasing (P<0.05), the expression of Osterix and osteocalcin was significantly increased after 1 day of incubation (P<0.05). However, compared with 0 μmol/L curculigoside treatment, the expression of osteopontin in MG63 cells was not significantly different after incubation with 1.0, 2.5, 5.0 μmol/L of curculigoside for 7 and 14 days (P>0.05). Compared with the blank group, the number of tibia osteoclasts in the osteoporosis model group increased. In the low-dose and high-dose groups of curculigoside, the tibia cortex was more continuous and the number of osteoclasts decreased. Compared with the blank group, the activity of oxygen in the osteoporosis model group was significantly increased (P<0.05), and superoxide dimutase and catalase were significantly decreased (P<0.05).@*Conclusions@#Curculigoside promotes the differentiation of MG63 cells by increasing the expression of osteoblast differentiation-related proteins, and has a certain therapeutic effect on dexamethasone-induced osteoporosis mice.
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Objective:To establish the quality standard for Erdingerxian mixture. Methods:Herba Violae and Cortex Phellodendri in Erdingerxian mixture were identified by TLC. Icariin and curculigoside were determined by HPLC. Results:The characteristic iden-tification by TLC was distinct and highly specific. The linear range of icariin was 5.77-184.50μg·ml-1(r=0.999 9),and the aver-age recovery was 96.55%(RSD=1.14%,n=9). The linear range of curculigoside was 12.36-395.50 μg·ml-1(r=1.000 0),and the average recovery was 100.58%(RSD=2.92%,n=9).Conclusion: The method is reliable, accurate, sensitive and specific, which can be used for the quality control of Erdingerxian mixture.
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Objective Curculiginis extract has good scavenging effects on oxygen free radicals, and there is no curculiginis extract for AD research at home.The aim of this study was to explore the influence of Curculigoside on the cognitive learning and memory functions of senile dementia model rats induced by D-galactose combined with β-protein amyloid β25-35 (Aa25 35) and the expression level of hippocampus B-cell lymphoma-2 (Bcl-2), Bax, estrogen receptor (ER) protein and mRNA.Methods Forty SD rats weredivided into the control group (equal saline was intraperitoneal injected for six weeks consecutively, and equal saline was injected to the bilateral hippocampus in the seventh week), the model group (galactose was intraperitoneal injected for six weeks consecutively, and 4 mmol/L Aβ25-35 was injected to the bilateral hippocampus in the seventh week), the low dosage group and the high dosage group randomly (the stomach irrigation of low dosage 24mg/kg or the high dosage 72mg/kg curculigoside was applied every day, for 8 weeks), 10 rats in each group.After the treatment , the Morris water maze was conducted to test the space learning and memory functions of rats;the flow cytometry method was conducted to analyze the neuronal apoptosis conditions;the western blot was conducted to detect Bcl-2, Bax and ER protein level;the Realtime PCR was conducted to detect Bcl-2, Bax and ER mRNA level.Results Morris water maze test results: Compared with the model group, the escape latency (EL) of the rats of the model group, the low dosage group and the high dosage group on the 2nd, the 3rd, the 4th and the 5th day significantly extends (P<0.01), and the high dosage group significantly shortens (P<0.05) than the low dosage group.Space exploration test results: Compared with the proportion of the distance swum within the quadrant by the rats in the model group in the total distance [(30.14±5.10)%], that of the low dosage group, the high dosage group and the control group [(41.98±4.78)%, (46.17± 9.31)% and (50.13±6.81)%] significantly increases (P<0.01);in addition, the high dosage group is higher than the low dosage group (P<0.05).Compared with the apoptosis rate of rates in the model group[(7.11±1.78)%], that of the high dosage group, the low dosage group and the control group[(5.53±1.45)%], (4.11±1.03)% and (3.08±1.33)% significantly decreases (P<0.05).Compared with the control group, the Bcl-2 and Bax protein expression of the model group significantly increases, and the cell Bcl-1 and Bax of cells significantly decreases (P<0.05).Compared with the model group, the Bcl-2 and ER protein expression as well as the Bcl-2/Bax of the high dosage group and the low dosage group significantly increase, and the Bax protein expression significantly decreases (P<0.05).Compared with the mRNA expression (1.00 and 1.00) of Bcl-2 and ER in the control group, the mRNA expression (1.31 and 1.98) of the Bcl-2 and Bax in the model group significantly increases (P<0.05);compared with the mRNA expression (1.13 and 1.08) of Bcl-2 and ER in the model group, the Bcl-2 (1.95 and 2.01) and mRNA expression (1.81 and 1.96) of ER of the high dosage group and the low dosage group significantly increase (P<0.01).Conclusion Curculigoside plays an improving role in space learning and memory functions of senile dementia model rats, with the mechanism may be related to the protection to the hippocampus neurons by restraining Bax expression and neuronal apoptosis through the up-regulation of hippocampus ER and Bcl-2 expressions.
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Objective:To study the extraction for the curculigoside and polysaccharide of Rhizoma Curculiginis. Methods: The optimum extraction was selected by the orthogonal design. The content of total curculigoside was determined by HPLC, The content of polysaccharide of Rhizoma Curculiginis was determined by spetrophotometry.Results: The extraction times affected significantly total curculigoside and Polysaccharide of Rhizoma Curculiginis. Conclusion: The optimum extraction is as follows: adding 8 times amount of water into Chinese medical materials, and boiling for 90 min each time, extracting three times in all.