Objective To observe the effect of transplanting the adipose-dervived stem cells(ADSCs) on free radical metabolism and immune function of rat aging model induced by D-galactose from fasiaology perspective;to explore a new method for anti-aging. Methods The ADSCs were cultured in vitro. Thirty male SD rats were randomly divided into 3 groups: normal control group(A), aging model group(B) and treat group(C). Ten rats in each group. Rats in B and C groups were injected D-galactose continually into make the sub-acute aging model rats.After 8-week injections of D-galactose;R3ats in group C were injected ADSCs(3×10~6/ml) through caudal vein. After 2-week transplantions of ADSCs, T-SOD, CuZn-SOD, MDA, NO, IL-2 and spleen index levels in serums of each group were detected and compared among the three groups. Results Compared with the A group, the SOD, NO, IL-2 level and spleen index in serum in group B decreased significantly, while the contents of MDA increased significantly. Compared with group B, the SOD, NO, IL-2 level and spleen index in serum in group C had been improved, and the contents of MDA decreased significantly. Conclusion Transplanting ADSCs can improve the antioxidant ability and strengthen the cellular immune function of aging rats.Further more, it can delay the ageing procedure induced by D-galactose in rats.

Objective To investigate the effect of in vitro high glucose stimulation on the expression of adiponectin receptor (adipoR) in human kidney proximal tubular cells.Methods The HK-2 cells were cultured in the low glucose DMEM culture medium containing 10% fetal bovine serum until the cells were adherent and 80% confluence. After cultured in the serum-free DMEM for 24 hours, these cells were stimulated with glucose-containing 1mg/ml, 2mg/ml, 4mg / ml, 6mg/ml, 8mg/ml serum-free DMEM for 48 hours. Then RT-PCR and western blot were used to analyze adipoR ( R1, R2) expression levels. The HK-2 cells were cultured respectively in high glucose (4mg/ml) , low glucose (1mg/ml) DMEM culture medium containing 10% fetal bovine serum to cultivate 0h, 12h, 24h, 48h, 72h, 96h, then RT - PCR was applied to analyze adipoR (R1, R2) mRNA expression levels semi-quantitatively. Results Two kinds of adiponectin receptor gene were both expressed in HK-2 cells, and the quantity of gene expression of adipoR1 (0. 63 ±0. 12) was 3. 9 times to adipoR2 (0. 16 ±0.03) , the difference was statistically significant ( P<0. 01). The different concentrations of glucose and different time of high glucose on HK-2 cells had no significant effect ( P>0. 05 ) on adipoR gene expression. Expression of adipoR 1 protein in HK-2 cells was detected by western blot, and it was not affected by glucose concentration ( P>0. 05).Conclusion adi-poR1 and adipoR2 gene were both expressed in HK-2 cells, and the adipoR1 was the major one, which suggested that adipoR1 played a more significant role in kidney disease. The expression of adipoRl/R2 of HK-2 cells was not affected by high glucose concentration.

BACKGROUND:The researches about the effect of retinoic acid on the proliferation of adipose-derived stem cells are rare, and the researches on the testosterone are mainly on the inhibition of cellaging. OBJECTIVE: To study the effects of retinoic acid and testosterone or combination on the cellcycle of adipose derived stem cells. METHODS:Adipose derived stem cells were isolated from adult female Sprague Dawley rats with 2 months age and cultured in vitro til passage 3 adipose derived stem cells, and then the 3rd passage adipose-derived stem cells were performed with adipogenic induction, osteogenic induction and surface marker identification. The cells were divided into six groups:(1) Control group;(2) 10-5 mol/L retinoic acid group;(3) Retinoic acid group;(4) 10-5 mol/L retinoic acid+testosterone group;(5) 10-6 mol/L retinoic acid+testosterone group;(6) Testosterone group. The adipose-derived stem cells in the control group were cultured with Dulbecco’s modified Eagle’s medium+10%fetal bovine serum culture medium, and the adipose-derived stem cells in the other five groups were induced with corresponding dose of retinoic acid and testosterone on the basis of control group. After cultured for 36 hours, the flow cytometry was used to detect the changes of cellcycle. RESULTS AND CONCLUSION:Compared with the control group, cellproportions in phase G 1 of 10-5 mol/L retinoic acid group and 10-6 mol/L retinoic acid group were increased significantly, and the cellproportions in phase S were decreased. Compared with control group, the cellproportion in phase G 1 of testosterone group was significantly reduced, and the cellproportion in phase S was increased. Compared with 10-5 mol/L retinoic acid group and 10-6 mol/L retinoic acid group, cellproportions in phase G 1 of 10-5 mol/L retinoic acid+testosterone group and 10-6 mol/L retinoic acid+testosterone group were reduced significantly and the cellproportions in phase S were increased. Retinoic acid can inhibit the cellcycle of adipose-derived stem cells in phase G 1 , and delay the process of the cellcycle from phase G1 to phase S;while testosterone can promote the cellcycle of adipose-derived stem cells from phase G1 to phase S;the combination induction of retinoic acid and testosterone can accelerate the process of the cellcycle of adipose-derived stem cells from phase G 1 to phase S.

OBJECTIVETo investigate whether adipose-derived stem cells (ADSCs) induced by retinoic acid (RA) in vitro express primordial germ cell marker alkaline phosphatase (ALP) and vasa.

METHODSADSCs were isolated from adult female SD rats and cultured in vitro. The third passage of ADSCs was identified by adipogenic differentiation, osteogenic differentiation and cell surface marker detection. The ADSCs were treated with 1×10(-5), 1×10(-6), or 1×10(-7) mol/L RA for 7 or 14 days, and the cellular expression of ALP was detected. vasa mRNA expression in ADSCs treated with 1×10(-5) mol/L RA for 7 days was detected using RT-PCR.

RESULTSThe OD value of ADSCs treated with 1×10(-5), 1×10(-6), or 1×10(-7) mol/L RA was 0.59∓0.04, 0.27∓0.07, and 0.15∓0.03 after a 7-day treatment, and was 0.42∓0.02, 0.34∓0.01, and 0.19∓0.02 after a 14-day treatment, respectively, demonstrating significantly enhanced ALP expression in RA-treated ADSCs compared with that in the control cells (0.07∓0.01 and 0.07∓0.01 at 7 and 14 days, respectively, P<0.01). The ADSCs showed a negative vasa mRNA expression after 1×10(-5) mol/L RA treatment for 7 days.

CONCLUSIONRA-induced ADSCs express ALP, a marker of primordial germ cells, but does not express the primordial germ cell marker vasa.

Adipose Tissue ; cytology ; Adult Stem Cells ; cytology ; enzymology ; Alkaline Phosphatase ; metabolism ; Animals ; Cell Differentiation ; Cells, Cultured ; Female ; Germ Cells ; cytology ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tretinoin ; pharmacology

Objective : To investigate the effects of hederagenin (HDG) on proliferation , migration , invasion and apoptosis of glioblastoma (GBM) cells and involved mechanism. Methods : Human GBM cell lines U87 , U251 and human brain glial cell line (HEB) were selected as the study subjects , and HDG 0 μmol/L ( or 0 mg/kg) was used as the control group. MTT , EdU staining and cell plate cloning were used to detect the effect of HDG on the proliferation of GBM cells. Trypan blue staining was used to detect GBM cell death affected by HDG. The effects of HDG on migration and invasion of GBM cells were detected by cell scratch and Transwell assay. To analyze the effects of HDG on apoptosis of GBM cells , apoptosis⁃related proteins Bcl⁃2 , Bax , p53 and cleaved caspase⁃3 were detected by Western blot. Mitochondrial potential change was detected by JC⁃10 staining , and apoptotic cell count was displayed by Annexin V ⁃FITC staining. The effect of HDG on tumor bearing in GBM was analyzed by xeno transplantation in BALB/C mice. Results : Compared with the control group (HDG 0 μmol/L) , HDG significantly inhibited the proliferation , migration and invasion of U87 and U251 cells , and they were dependent on the use dose of HDG. Trypan blue staining showed that HDG obviously increased death number of GBM cells. The mitochondrial potential of GBM cells was remarkedly decreased , the number of apoptotic GBM cells obviously increased , the expressions of apoptosis⁃related proteins p53 , Bax , cleaved⁃caspase3 were up⁃regulated and Bcl⁃2 was down⁃regulated by HDG in U87 and U251 cells. HDG significantly inhibited the size of subcutaneous GBM , the Ki67 positive rate of GBM cells and caused a large number of GBM cells to die in BALB/C mice. HDG had no obvious toxic effect on human HEB cells and the liver of tumor⁃bearing mice. Conclusion HDG can significantly inhibit the proliferation , migration and invasion of GBM cells and induce the apoptosis of them. The mechanism of HDG induced apoptosis of GBM cells may be through mitochondrial damage and regulation of p53 and Bcl⁃2/Bax expression.