OBJECTIVE:To optimize the extraction technology of asiaticosides from Centella asiatica. METHODS:Using to-tal amounts of asiaticoside and hydroxy asiaticoside as investigation indexes,single factor and orthogonal test were used to investi-gate the enzyme amount,enzymolysis time,enzymolysis temperature,ethanol volume fraction,liquid material ratio and ultrasonic extraction time and optimize the extraction technology of asiaticosides from C. asiatica by ultrasonic-enzyme method,and verifica-tion test was conducted. RESULTS:Optimal extraction technology was as follow as cellulose dosage of 12 mg/g,10-fold liquid ma-terial ratio added into 60% ethanol,enzyme hydrolysis for 60 min at 60 ℃,ultrasonic assisted extraction for 50 min. Average ex-traction rate of total asiaticosides was 1.92%(RSD=1.83%,n=3)in verification test. CONCLUSIONS:Ultrasonic-enzyme meth-od is stable and feasible for the extraction technology of asiaticosides from C. asiatica.

Objective To investigate the optimal conditions for establishing insulin-resistant 3T3-L1 adipocytes. Methods Dexamethason (DEX), 3-isobutyl-methylxanthine (IBMX) and different concentrations of insulin (10-8, 10-7, and 10-6 mol · L-1) were used to induce 3T3-L1 preadipocytes into mature adipocytes identified by oil red O staining. We established insulin-resistant 3T3-L1 adipocytes cell model (IR-3T3-L1) by exposing the cells to 1μmol· L-1 DEX, and the changes of glucose concen-tration in the cell culture were determined by glucose oxidase-peroxidase (GOD-POD) assay. Results Treatment of 3T3-L1 cells with DEX, IBMX and 10-6 mol· L-1 insulin for 9 days resulted in the differentiation of>90%of the cells into mature adipocytes. IR-3T3-L1 cells cultured for 96 h in the culture media containing 1 μmol · L-1 DEX showed significantly increased glucose consumption (P=0.0003) as compared with the control group at 36 h (P<0.001). Conclusion 3T3-L1 cells can be induced into mature adipocytes by exposure to 1μmol· L-1 DEX, 0.5 mmol· L-1 IBMX and 10-6 mol· L-1 insulin. A 96 h exposure to 1μmol· L-1 DEX can induce 3T3-L1 adipocytes to acquire insulin resistance that can be maintained for 36 h.

Objective To investigate the optimal conditions for establishing insulin-resistant 3T3-L1 adipocytes. Methods Dexamethason (DEX), 3-isobutyl-methylxanthine (IBMX) and different concentrations of insulin (10-8, 10-7, and 10-6 mol · L-1) were used to induce 3T3-L1 preadipocytes into mature adipocytes identified by oil red O staining. We established insulin-resistant 3T3-L1 adipocytes cell model (IR-3T3-L1) by exposing the cells to 1μmol· L-1 DEX, and the changes of glucose concen-tration in the cell culture were determined by glucose oxidase-peroxidase (GOD-POD) assay. Results Treatment of 3T3-L1 cells with DEX, IBMX and 10-6 mol· L-1 insulin for 9 days resulted in the differentiation of>90%of the cells into mature adipocytes. IR-3T3-L1 cells cultured for 96 h in the culture media containing 1 μmol · L-1 DEX showed significantly increased glucose consumption (P=0.0003) as compared with the control group at 36 h (P<0.001). Conclusion 3T3-L1 cells can be induced into mature adipocytes by exposure to 1μmol· L-1 DEX, 0.5 mmol· L-1 IBMX and 10-6 mol· L-1 insulin. A 96 h exposure to 1μmol· L-1 DEX can induce 3T3-L1 adipocytes to acquire insulin resistance that can be maintained for 36 h.

Stem cells can be differentiated into many kinds of somatic cells under defined culture conditions. In addition, the homing possess can be partially imitated by co-culture of stem cells with mature somatic cells. Regarding the importance of clinical application of adipose-derived stem cells (ADSCs), our review first introduced the sources and signs of ADSCs, and then the current knowledge of ADSCs co-culture technology, including drug and chemical induced culture, two-dimensional (2D) and three-dimensional (3D) co-culture, mechanisms of ADSCs differentiation, and application development in recent years in details. Finally, we also addressed prospects of ADSCs.
Adipose Tissue ; cytology ; Animals ; Cell Differentiation ; drug effects ; Cells, Cultured ; Coculture Techniques ; methods ; Humans ; Stem Cells ; cytology ; Tissue Engineering

The aim of this study was to evaluate the efficacy of Bletilla striata polysaccharide on diabetes mellitus ulcers. Diabetes mellitus animal model was established by single ip injection of streptozotocin (STZ, 50 mg x kg(-1)) with the criteria of blood glucose > or = 16.7 mmol x L(-1) after 72 h. 4 weeks after STZ injection, each animal received two full thickness incisional wounds (1.8 cm in diameter). The wounds then were divided into B. striata polysaccharide group and PBS group. Wound closure rate, fibroblast (FB) infiltration, hydroxyproline (OHP) content and myeloperoxidase (MPO) levels were examined on day 3, 7, 14, 21 post wound. The treatment of B. striata polysaccharide significantly facilitated diabetes mellitus ulcers healing compared to PBS group. Histological analysis showed that B. striata polysaccharide markedly increased inflammatory cell infiltration in wound area. The herb also strongly evaluation of FB, OHP demonstrated a significantly increased in B. striata polysaccharide group. B. striata polysaccharide group promoted wound closure by means of enhanced inflammatory cell infiltration and re-epithelialization, and the promotion of FB and OHP levels.
Animals ; Diabetes Complications ; drug therapy ; Diabetes Mellitus, Experimental ; chemically induced ; drug therapy ; Drugs, Chinese Herbal ; administration & dosage ; Fibroblasts ; drug effects ; metabolism ; Hydroxyproline ; drug effects ; metabolism ; Male ; Peroxidase ; drug effects ; metabolism ; Plant Extracts ; administration & dosage ; Polysaccharides ; administration & dosage ; Rats ; Skin Ulcer ; drug therapy ; Wound Healing ; drug effects

OBJECTIVETo study the anti-inflammatory mechanism of total saponins from Semen Nigellae (TSSN).

METHODIFN-gamma plus LPS stimulated RAW 264. 7 macrophage has been used as inflammatory experimental model. Griess reaction for nitric oxide production, FRAP assay for total antioxidant capacity, RT-PCR for mRNA expression and Western blot for protein expression examination were performed.

RESULTTSSN inhibited NO production in a dose-dependent manner. The gene and protein expression of iNOS were also suppressed by the herb extract. TSSN treatment significantly attenuated mRNA of inflammatory mediators such as COX-2, IL-1beta, IL-6 while increased PPAR-gamma gene and protein expression. Furthermore, phosphorylation of ERK (p-ERK) was markedly inhibited by TSSN.

CONCLUSIONTSSN suppressed pro-inflammatory mediators such as COX-2, IL-1beta, IL-6 and increased anti-inflammatory mediator PPAR-gamma expression. Meanwhile, TSSN inhibited over production of NO and iNOS expression through ERK/MAPK pathway.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Cells, Cultured ; Cyclooxygenase 2 ; metabolism ; Humans ; Inflammation ; chemically induced ; Inflammation Mediators ; metabolism ; Interleukin-6 ; metabolism ; Lipopolysaccharides ; pharmacology ; Macrophages ; drug effects ; enzymology ; metabolism ; Mice ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Phosphorylation ; drug effects ; Saponins ; pharmacology ; Signal Transduction