OBJECTIVETo investigate the role of mitochondria in TBMS1-medited apoptosis of human cervical carcinoma HeLa cell line.

METHODMitochondrial transmembrane potentia (deltapsim) was assayed by flow cytometry. Apoptotic induction by TBMS1 was determined by gel electrophoresis of fragmented DNA. Cytochrome c (Cyt c) was detected by Western blotting.

RESULTThe results showed that TBMS1 induced apoptosis in HeLa cells, that TBMS1 decreased deltapsim and facilitated Cyt c release, and that TBMS1 induced apoptosis of HeLa cells dose-and time-dependently in accordance with increase of cytosolic Cyt c. TBMS1 had direct effect on isolated rat mitochondria which induced a time- and dose-dependent release of Cyt c from mitochondria. The results also showed that CsA protected partly HeLa cells from the effect of TBMS1.

CONCLUSIONThe mitochondrial apoptosis pathway has effects on TBMS1 induced human cervical carcinoma HeLa cell line apoptosis.

Animals ; Apoptosis ; drug effects ; Cucurbitaceae ; chemistry ; Cyclosporine ; pharmacology ; Cytochromes c ; metabolism ; Dose-Response Relationship, Drug ; HeLa Cells ; Humans ; Male ; Membrane Potentials ; drug effects ; Mitochondria, Liver ; physiology ; secretion ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Saponins ; administration & dosage ; isolation & purification ; pharmacology ; Triterpenes ; administration & dosage ; isolation & purification ; pharmacology

We studied the alteration of the maize straw apoplast proteins in the process of preservation, and analyzed the effects of apoplast proteins on Penicillum expansum cellulase activities. The results show that: the extractable apoplast proteins are gradually decreased during the preservation of maize straw. Meanwhile, their synergistic effects on P. expensum cellulose are also attenuated. The apoplast proteins extracted from fresh maize straw possess endogenous EG activities, which is unstable and completely vanished after 6 months preservation. The apoplast proteins from the preserved straw exhibit significant synergistic effect on FPA, cotton lyase and beta-glucosidase. The maximal synergistic values are 95.32%, 102.06% and 96.6%, respectively. But interestingly, they inhibit the CMCase activity (max. 49.52%). Apoplast proteins show distinctive synergy with betaG and EG, but have no effect on CBH activity. After eliminating the effect of endogenous EG, the apoplast proteins from fresh maize straw have enhanced synergistic or inhibiting effects on FPA, Cotton lyase, betaG and CMCase than those extracted from the preserved straw. Based on our observation, the apoplast proteins play important roles in regulating the cellulase activities. The detailed analysis of the related mechanisms will greatly benefit the studies of the natural biomaterials hydrolysis.
Cellulase ; metabolism ; Penicillium ; enzymology ; Plant Proteins ; metabolism ; Plant Stems ; metabolism ; Zea mays ; metabolism

OBJECTIVETo observe the ultrastructural changes of HeLa cells in response to tubeimoside I (TBMS1) treatment and the protective effect of cyclosporine A (CsA), and explore the role of intrinsic apoptosis pathway in TBMS1-induced HeLa cell apoptosis.

METHODSHeLa cells were treated with TBMS1 (10-50 micromo/L) alone or in combination with 2 micromol/L CsA for 12 and 24 h and observed with transmission electron microscope (TEM) for the ultrastructural changes of the cells.

RESULTSTBMS1 induced apoptosis of HeLa cells in a concentration- and time-dependant manner. Under TEM, the treated cells progressively shrunk and the intercellular space widened with loss of microvillus, mitochondrial swelling, rough endoplasmic reticulum enlargement, chromatin condensation, nuclear shrinkage and nuclear pyknosis as TBMS1 concentration increased. At low concentrations, CsA offered partial protection of the mitochondria from TBMS1-induced damage whereas high-concentration CsA did not.

CONCLUSIONTBMS1 induces ultrastructural changes typical for apoptosis of the HeLa cells, which provides morphological evidence for the role of intrinsic apoptosis pathway in TBMS1-induced apoptosis.

Apoptosis ; drug effects ; Cell Nucleus ; drug effects ; ultrastructure ; Cyclosporine ; pharmacology ; Dose-Response Relationship, Drug ; Endoplasmic Reticulum, Rough ; drug effects ; ultrastructure ; Female ; HeLa Cells ; Humans ; Immunosuppressive Agents ; pharmacology ; Microscopy, Electron, Transmission ; Mitochondrial Swelling ; drug effects ; Saponins ; pharmacology ; Time Factors ; Triterpenes ; pharmacology ; Uterine Cervical Neoplasms ; pathology ; ultrastructure

Objective To investigate a possible protective effect of sodium ferulate (SF) on monosodium glutamate (MSG)-induced neurotoxicity in adult mice. Methods Sixty mice were randomly divided into control, SF, MSG, and MSG+SF [20,40,80mg/(kg·d)] groups, n=10. The animals in MSG group received intragastric (ig) administration of MSG (2.0g/(kg·d)], the animals in the MSG+SF groups received simultaneously ig administration of MSG [2.0 g/(kg·d)] and intraperitoneal (ip) administration of SF [20,40,80mg/(kg·d)], the animals in SF group received ip administration of SF [40mg/ (kg·d)], and the animals in control group received ig and ip administration of normal saline, respectively, once-daily for 10d. On day 1 after the last ig administration of MSG or (and) SF the behavioural tests (test of Y-maze discrimination learning and open field test) were performed, and on day 4 after the treatment of MSG or (and) SF the histopathology of the animal brains was studied to analyze the MSG-induced functional and morphological changes and the possible protective effect of SF. Results The correct responses of Y-maze test on day 6 after the last administration of MSG and/or SF in MSG-treated group (13.83/20) were significantly less than those in control (16.42/20)(P<0.01), and those in MSG[2.0g/(kg·d)]+SF[40mg/(kg·d)]-treated mice (16.30/20) were close to those in control (P>0.05). Examination of histopathology displayed MSG-treated hippocampal lesions characterized by intracellular edema, degeneration and necrosis of neurons, and hyperplasia, and the hippocampal lesion did not appear in the MSG [2.0g/(kg·d)]+SF[40mg/(kg·d)]-treated mice. Conclusions SF partially countered the behavior disorders and hippocampal lesions induced by MSG; therefor, SF has a potent neuroprotection against MSG-induced neurotoxicity in adult mice.

OBJECTIVETo investigate a possibility of repairing damaged brain by intracerebroventricular transplantation of neural stem cells (NSCs) in the adult mice subjected to glutamate-induced excitotoxic injury.

METHODSMouse NSCs were isolated from the brains of embryos at 15-day postcoitum (dpc). The expression of nestin, a special antigen for NSC, was detected by immunocytochemistry. Immunofluorescence staining was carried out to observe the survival and location of transplanted NSCs. The animals in the MSG + NSCs group received intracerebroventricular transplantation of NSCs (approximately 1.0 x 10(5) cells) separately on day 1 and day 10 after 10-d MSG exposure (4.0 g/kg per day). The mice in control and MSG groups received intracerebroventricular injection of Dulbecco's minimum essential medium (DMEM) instead of NSCs. On day 11 after the last NSC transplantation, the test of Y-maze discrimination learning was performed, and then the histopathology of the animal brains was studied to analyze the MSG-induced functional and morphological changes of brain and the effects of intracerebroventricular transplantation of NSCs on the brain repair.

RESULTSThe isolated cells were Nestin-positive. The grafted NSCs in the host brain were region-specifically survived at 10-d post-transplantation. Intracerebroventricular transplantation of NSCs obviously facilitated the brain recovery from glutamate-induced behavioral disturbances and histopathological impairs in adult mice.

CONCLUSIONIntracerebroventricular transplantation of NSCs may be feasible in repairing diseased or damaged brain tissue.

Animals ; Cell Count ; Disease Models, Animal ; Embryo, Mammalian ; Glutamic Acid ; toxicity ; Injections, Intraventricular ; methods ; Intermediate Filament Proteins ; metabolism ; Mice ; Mice, Inbred Strains ; Nerve Tissue Proteins ; metabolism ; Nestin ; Neurons ; physiology ; Neurotoxicity Syndromes ; etiology ; pathology ; surgery ; Stem Cell Transplantation ; methods ; Stem Cells ; physiology ; Time Factors

Child ; Humans ; von Willebrand Disease, Type 2