PURPOSE: This study aimed to elucidate the molecular mechanisms of the anti-pancreatic fibrosis effects of matrine in rats. MATERIALS AND METHODS: Trinitrobenzene sulfonic acid was administrated to rats to establish a pancreatic fibrosis model. Rats were divided into four groups: Control, Sham, Model, and Matrine (n=8). Hematoxylin-eosin staining, Masson staining, and Azan staining were performed to evaluate pancreatic fibrosis. Expression of transforming growth factor-β1 (TGF-β1), α-smooth muscle actin (α-SMA), and collagen I in pancreatic tissues was evaluated by immunohistochemical staining. mRNA and protein levels of TGF-β receptor 1 (TβR1), TβR2, and Smad2 in pancreatic tissues were determined by RT-PCR and Western blot, respectively. RESULTS: In the model group, hyperplasia of glandules around the glandular ducts, mitochondrial swelling of acinous cells, and severe fibrosis were found. Interestingly, in the Matrine group, mitochondrial swelling was only found in a small number of acinous cells, and the fundamental structures of pancreatic tissues were intact. Moreover, pancreatic fibrosis was markedly alleviated. Comparing to the Sham group, expression of α-SMA, TGF-β1, and collagen I was sharply elevated in the Model group (p < 0.05); however, their expressions were much lower in the Matrine group, compared to the Model group (p < 0.05). Compared with the Sham group, mRNA and protein levels of Smad2, TβR1, and TβR2 in the Model group were notably raised (p < 0.05). However, their high expression was significantly downregulated in the Matrine group (p < 0.05). CONCLUSION: Matrine suppressed pancreatic fibrosis by regulating TGF-β/Smad signaling in rats.
Acinar Cells ; Actins ; Animals ; Blotting, Western ; Collagen ; Fibrosis* ; Hyperplasia ; Mitochondrial Swelling ; Rats* ; RNA, Messenger ; Signal Transduction

BACKGROUND: Intravenous palonosetron-HCl, a second-generation antagonist of selective serotonin type 3 (5-HT3) receptors, can prevent chemotherapy-induced nausea and vomiting (CINV) and postoperative nausea and vomiting (PONV). 5-HT3 receptors are abundant in the lower brainstem and the substantia gelatinosa of the spinal cord, which provides a theoretical rationale for neuraxial administration of 5-HT3 receptor antagonists for CINV, PONV, and opioid-induced nausea and vomiting. However, there are no reports of neuraxial administration of palonosetron-HCl. Before neuraxial administration of a drug is accepted for clinical use, its safety must be proven. This study was conducted to determine whether neuraxial administration of palonosetron-HCl produces neurologic injury. METHODS: Male Sprague-Dawley rats under general anesthesia were catheterized intrathecally and the catheter tip was advanced caudally to the L1 vertebra. After 7 days, 20 µl of normal saline (N group, n = 6) or 20 µl (1 µg) of palonosetron-HCl (P group, n = 6) were injected intrathecally once per day for 2 weeks. Neurotoxic changes were evaluated by light microscopy (LM) and electron microscopy (EM) of the spinal cord. Behavioral changes were also evaluated in both groups. RESULTS: One of the N group rats and three of the P group rats demonstrated abnormal behavior during intrathecal drug injection, but otherwise their behavior was normal. The spinal cords of the N group did not have any abnormal findings by LM or EM. The spinal cords of the P group had multiple vacuoles in the white matter by LM, especially in the dorsal funiculus, and EM revealed myelin, axonal, and mitochondrial swelling. CONCLUSIONS: Results suggest that chronic intrathecal administration of palonosetron-HCl produced microscopic morphologic changes in the spinal cords of rats.
Anesthesia, General ; Animals ; Axons ; Brain Stem ; Catheters ; Humans ; Injections, Spinal ; Male ; Microscopy ; Microscopy, Electron ; Mitochondrial Swelling ; Myelin Sheath ; Nausea ; Postoperative Nausea and Vomiting ; Rats* ; Rats, Sprague-Dawley ; Receptors, Serotonin, 5-HT3 ; Serotonin ; Spinal Cord* ; Spine ; Substantia Gelatinosa ; Vacuoles ; Vomiting ; White Matter

PURPOSE: Cell-in-cell structures are created by one living cell entering another homotypic or heterotypic living cell, which usually leads to the death of the internalized cell, specifically through caspase-dependent cell death (emperitosis) or lysosome-dependent cell death (entosis). Although entosis has attracted great attention, its occurrence is controversial, because one cell line used in its study (MCF-7) is deficient in caspase-3. METHODS: We investigated this issue using MCF-7 and A431 cell lines, which often display cell-in-cell invasion, and have different levels of caspase-3 expression. Cell-in-cell death morphology, microstructures, and signaling pathways were compared in the two cell lines. RESULTS: Our results confirmed that MCF-7 cells are caspase-3 deficient with a partial deletion in the CASP-3 gene. These cells underwent cell death that lacked typical apoptotic properties after staurosporine treatment, whereas caspase-3-sufficient A431 cells displayed typical apoptosis. The presence of caspase-3 was related neither to the lysosome-dependent nor to the caspase-dependent cell-in-cell death pathway. However, the existence of caspase-3 was associated with a switch from lysosome-dependent cell-in-cell death to the apoptotic cell-in-cell death pathway during entosis. Moreover, cellular hypoxia, mitochondrial swelling, release of cytochrome C, and autophagy were observed in internalized cells during entosis. CONCLUSION: The occurrence of caspase-independent entosis is not a cell-specific process. In addition, entosis actually represents a cellular self-repair system, functioning through autophagy, to degrade damaged mitochondria resulting from cellular hypoxia in cell-in-cell structures. However, sustained autophagy-associated signal activation, without reduction in cellular hypoxia, eventually leads to lysosome-dependent intracellular cell death.
Apoptosis ; Autophagy ; Caspase 3* ; Cell Death ; Cell Hypoxia ; Cell Line ; Cytochromes c ; Entosis ; MCF-7 Cells* ; Mitochondria ; Mitochondrial Swelling ; Staurosporine

OBJECTIVETo observe the impairing effects of triptolide on liver mitochondria in isolated rat-liver mitochondria and human normal liver HL7702 cell line.

METHODSRat-liver mitochondria were isolated from adult female Sprague-Dawley (SD) rats. Liver mitochondria were incubated with 0, 1.25, 2.5, 5 and 10 μmol/L triptolide for detecting mitochondrial swelling and with 0, 2.5, 5 and 10 μmol/L triptolide for mitochondrial permeability transition pore (MPTP) activity. Mitochondrial swelling was estimated by measuring the apparent absorbance change during 600 s in the mitochondrial suspensions at 520 nm with a mitochondrial swelling examining kit. The effect of triptolide on MPTP was determined with a fluorescence detection kit by detecting the fluorescence intensity at an excitation wavelength of 488 nm emitted at 527 nm. Human normal liver HL7702 cells were treated without or with 0.02, 0.1 and 0.5 μmol/L triptolide for 24 h for analyzing mitochondrial transmembrane potential (Δψm) and reactive oxygen species (ROS). Δψm was measured using the fluorescent probe 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolylcarbocyanine iodide (JC-1). ROS was measured using fluorescent probe 2',7'-dichlorofluorescin diacetate (DCFH-DA). The cells were harvested and dyed with JC-1 and DCFH-DA, and analyzed by flow cytometry, respectively.

RESULTSIncubation of isolated mitochondria with triptolide results in swollen mitochondria in a concentration-dependent manner. Moreover, triptolide significantly activated mitochondrial permeability transition at 5 and 10 μmol/L (P<0.05 and P<0.01). When HL7702 cells were exposed to a various concentration triptolide for 24 h, mitochondrial membrane depolarization and increase of ROS were caused by triptolide in a concentration-dependent manner. Triptolide significantly induced the mitochondrial membrane depolarization at 0.1 and 0.5 μmol/L (P<0.05 and P<0.01) and the increase of ROS at 0.1 and 0.5 μmol/L (P<0.05 and P<0.01).

CONCLUSIONTriptolide could induce mitochondrial impairment, which may be one of the mechanisms by which hepatotoxicity occurs.

Animals ; Cell Line ; Diterpenes ; chemistry ; pharmacology ; Epoxy Compounds ; chemistry ; pharmacology ; Female ; Humans ; Membrane Potential, Mitochondrial ; drug effects ; Mitochondria, Liver ; drug effects ; metabolism ; Mitochondrial Membrane Transport Proteins ; metabolism ; Mitochondrial Swelling ; drug effects ; Phenanthrenes ; chemistry ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species ; metabolism

BACKGROUND: Role of cytochrome c (Cyt c) is an apoptogenic agent under certain conditions. The mitochondrial permeability transition pore (MPTP) plays an important role in cell death since it opens, leading to mitochondrial swelling and release of Cyt c, which initiates apoptosis. By inhibiting the opening of MPTP, cyclosporine A (CSA) may contribute to maintaining mitochondrial homeostasis. We investigate the effects of the partial sciatic nerve injury (PSNI)-induced neuropathic pain model on mitochondrial Cyt c release and the effects of CSA on neuroprotection by mitochondrial stabilizing activity in PSNI rats. METHODS: Rats were assigned to two groups that received different operations (Group P; PSNI operation, Group S; sham operation). The changes of cyt c and GABAergic neuron were evaluated in the spinal cord tissue. After which, PSNI rats randomly received CSA (Group C) or saline (Group S), and the changes of mechanical thresholds with Cyt c and GABAergic neuron were checked. RESULTS: PSNI in rats increased the release of cytosolic Cyt c. However, GABAergic cells were not decreased in the spinal cord level on the ipsilateral side to the PSNI. The second experiment reveal a reduction in Cyt c release, using CSA in PSNI model. Rats receiving CSA were afforded the antiallodynia without decrease of GABAergic cell. CONCLUSIONS: The Cyt c probably contributes to nerve dysfunction after PSNI. PSNI induced neuropathic pain was profoundly linked to mitochondrial stabilization. Thus, the potent neuroprotector, CSA, might produce antiallodynia through its capability to inhibit the opening of MPTP.
1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine ; Animals ; Apoptosis ; Cell Death ; Cyclosporine ; Cytochromes ; Cytochromes c ; Cytosol ; GABAergic Neurons ; Homeostasis ; Hyperalgesia ; Mitochondrial Membrane Transport Proteins ; Mitochondrial Swelling ; Neuralgia ; Permeability ; Rats ; Salicylamides ; Sciatic Nerve ; Spinal Cord

OBJECTIVETo study effects of three pentacyclic triterpenoids, oleanolic acid (OA), ursolic acid (UA) and asiatic acid (AA) on Ca(2+)-induced liver mitochondrial permeability transition (MPT).

METHODEffects of three compounds on liver MPT induced by Ca2+ were assessed by measuring the change in mitochondrial swelling, mitochondrial membrane potential and release of matrix Ca2+ in vitro.

RESULTObvious mitochondrial swelling, loss of mitochondrial membrane potential and release of matrix Ca2+ occurred after the addition of 50 micromol x L(-1) Ca2+. However, preincubation with 50 mg x L(-1) OA, UA or AA significantly blocked the above changes. In addition, it was also found that there are differences in the inhibitions of three compounds on liver MPT induced by Ca2+.

CONCLUSIONThree pentacyclic triterpenoids, OA, UA and AA, have significant mitochondrial protection through blocking on liver MPT and the inhibition on liver MPT of AA is stronger than that of UA and OA.

Animals ; Calcium ; metabolism ; pharmacology ; Membrane Potential, Mitochondrial ; drug effects ; Mice ; Mice, Inbred ICR ; Mitochondria, Liver ; drug effects ; metabolism ; Mitochondrial Membrane Transport Proteins ; drug effects ; Mitochondrial Swelling ; drug effects ; Pentacyclic Triterpenes ; pharmacology

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

OBJECTIVETo investigate the effects of resveratrol (Res) on mitochondrial opening and Ca(2+)-induced Ca(2+) release (CICR) from rat liver cell mitochondria mediated by Ca(2+).

METHODSWistar rat liver cell mitochondria was extracted and Res-induced mitochondrial swelling was assessed spectrophotometrically at 540 nm to examine the permeability transition pore (PTP) opening. The membrane potential changes of Res-treated mitochondria were measured with fluorescence spectrophotometery. Ca(2+) uptake and release by the mitochondria was determined by absorbance change of arsenazo III at 685-675 nm monitored by dual wavelength spectrophotometry.

RESULTSRes promoted Ca(2+)-mediated PTP opening, and this effect was completely inhibited by CsA and lowered by trifluoperazine. CICR accelerated by Res treatment was completely blocked by ruthenium red and partly by trifluoperazine.

CONCLUSIONRes can promote PTP opening by inducing CICR, which may be one of the pathways that Res induces cell apoptosis.

Animals ; Calcium ; metabolism ; Cells, Cultured ; Female ; Hepatocytes ; cytology ; metabolism ; Mitochondria, Liver ; drug effects ; metabolism ; Mitochondrial Membrane Transport Proteins ; metabolism ; Mitochondrial Swelling ; drug effects ; Rats ; Rats, Wistar ; Stilbenes ; pharmacology

OBJECTIVETo elvaulate the antioxidant activity of the pigment of Lycium ruthenicum.

METHODThe antioxidant activities were measured by the effects of the reducing ability, scavenging DPPH. H2O2-induced hemolysis of mice erythrocyte, serum resistance of reactive oxygen species, content of MDA in liver tissue, and swelling effect of mitochondria in liver tissue.

RESULTThe pigment of L. ruthenicum could scaveng DPPH* remarkably with IC50 0.164 mg x mL(-1), inhibitte hemolysis of mice erythrocyte evidently with IC50 0.112 mg x mL(-1). The resistant of reactive oxygen species was enhanced by the tested substances, simultanously. The concentration of MDA of peroxidation of lipid in mice liver could be reduced, and the swelling of mice liver mitochondria alse be restrained.

CONCLUSIONThe pigment of L. ruthenicum has antioxidant activity in tested concentration.

Animals ; Antioxidants ; pharmacology ; Biphenyl Compounds ; metabolism ; Erythrocytes ; drug effects ; Free Radical Scavengers ; pharmacology ; Hemolysis ; drug effects ; Hydrazines ; metabolism ; Lipid Peroxidation ; drug effects ; Liver ; metabolism ; Lycium ; chemistry ; Malondialdehyde ; metabolism ; Mice ; Mitochondria, Liver ; pathology ; Mitochondrial Swelling ; drug effects ; Phenols ; isolation & purification ; pharmacology ; Picrates ; Pigments, Biological ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Reactive Oxygen Species

OBJECTIVEThe aim of this study was to investigate the mechanisms of C. minima in the treatment of allergic rhinitis.

METHODAn allergic rhinitis animal model induced by ragweed pollen was established. After treatment with an active extract of C. minima, histopathological changes in the nasal mucosa of guinea pig were observed by transmission electron microscope.

RESULTIn the allergeic rhinitis model group, there appear a large number of lysosomes in the nasal epithelium with organelles vacuolated and nucleus deformed. Cells in the proper lamina of connective tissue were disarranged with organelles damaged, and there was also infiltration of eosinophils and mast cells in the connective tissue. However, in the treatment group receiving C. minima extract, the pathological changes mentioned above were significantly decreased.

CONCLUSIONC. minima is effective in treating allergic rhinitis.

Animals ; Asteraceae ; chemistry ; Epithelium ; ultrastructure ; Female ; Guinea Pigs ; Lysosomes ; drug effects ; Male ; Mitochondrial Swelling ; drug effects ; Nasal Mucosa ; pathology ; ultrastructure ; Oils, Volatile ; isolation & purification ; pharmacology ; Phytotherapy ; Plants, Medicinal ; chemistry ; Rhinitis, Allergic, Seasonal ; drug therapy ; pathology