aluminum-tetrasulfophthalocyanine and Necrosis

Oxidative stress is the reason of diverse neuropathological processes. Photodynamic therapy (PDT), an effective inducer of oxidative stress, is used for cancer treatment, including brain tumors. We studied the role of various signaling pathways in photodynamic injury and protection of single neurons and satellite glial cells in the isolated crayfish mechanoreceptor. It was photosensitized with alumophthalocyanine Photosens in the presence of inhibitors or activators of various signaling proteins. PDT eliminated neuronal activity and killed neurons and glial cells. Inhibitory analysis showed the involvement of protein kinases Akt, glycogen synthase kinase-3β (GSK-3β), mammalian target of rapamycin (mTOR), mitogen-activated protein kinase kinases 1 and 2 (MEK1/2), calmodulin, calmodulin-dependent kinase II (CaMKII), adenylate cyclase, and nuclear factor NF-κB in PDT-induced necrosis of neurons. Nitric oxide (NO) and glial cell-derived neurotrophic factor (GDNF) reduced neuronal necrosis. In glial cells, protein kinases Akt, calmodulin, and CaMKII; protein kinases C and G, adenylate cyclase, and p38; and nuclear transcription factor NF-κB also mediated PDT-induced necrosis. In contrast, NO and neurotrophic factors nerve growth factor (NGF) and GDNF demonstrated anti-necrotic activity. Phospholipase Cγ, protein kinase C, GSK-3β, mTOR, NF-κB, mitochondrial permeability transition pores, and NO synthase mediated PDT-induced apoptosis of glial cells, whereas protein kinase A, tyrosine phosphatases, and neurotrophic factors NGF, GDNF, and neurturin were involved in protecting glial cells from photoinduced apoptosis. Signaling pathways that control cell survival and death differed in neurons and glia. Inhibitors or activators of some signaling pathways may be used as potential protectors of neurons and glia from photooxidative stress and following death.

Topics: Animals; Apoptosis; Astacoidea; Enzyme Inhibitors; In Vitro Techniques; Indoles; Light; Mechanoreceptors; Necrosis; Nerve Growth Factors; Nerve Tissue Proteins; Neuroglia; Neurons; NF-kappa B; Nitric Oxide; Organ Specificity; Organometallic Compounds; Oxidative Stress; Phospholipase C gamma; Phosphoprotein Phosphatases; Photochemotherapy; Protein Kinases; Radiation-Sensitizing Agents; Signal Transduction

Photodynamic therapy (PDT) is a technique that can be used as a complementary therapy in cancer treatment combined with other therapeutic modalities. Quercetin (QCT) is known to be effective in the treatment of cancer, by reducing the cell viability of different cancer cell lines. This study aimed to evaluate the influence of different concentrations of QCT in PDT on the viability, mitochondrial membrane potential and induction of apoptosis/necrosis in the human larynx carcinoma cells (HEp-2). The HEp-2 cells were treated with aluminum phthalocyanine tetrasulfonate (AlPcS4) and QCT and subsequently irradiated with a diode laser light (685 nm, 35 mW, 4.5 J/cm(2)). The results demonstrated that treatment of HEp-2 cells with high concentrations of QCT (at least 50 μM) reduced cell viability. This response was enhanced in cells subjected to PDT supplemented with high concentrations of QCT. In addition, was observed decrease in the mitochondrial membrane potential and characteristics of late apoptosis and/or initial necrosis process. QCT at concentrations from 50 μM improves PDT-induced cytotoxicity by significantly reducing cell viability by apoptosis and/or necrosis, and mitochondrial membrane potential of Hep-2 cells.

Topics: Antineoplastic Agents; Apoptosis; Cell Line, Tumor; Cell Survival; Fluorescent Dyes; Humans; Indoles; Laryngeal Neoplasms; Larynx; Membrane Potential, Mitochondrial; Necrosis; Organometallic Compounds; Photochemotherapy; Photosensitizing Agents; Quercetin

Epigenetic processes are involved in regulation of cell functions and survival, but their role in responses of neurons and glial cells to oxidative injury is insufficiently explored. Here, we studied the role of DNA methylation and histone deacetylation in reactions of neurons and surrounding glial cells to photodynamic treatment that induces oxidative stress and cell death. Isolated crayfish stretch receptor consisting of a single mechanoreceptor neuron surrounded by glial cells was photosensitized with aluminum phthalocyanine Photosens that induced neuron inactivation, necrosis of the neuron and glia, and glial apoptosis. Inhibitors of DNA methylation 5-azacytidine and 5-aza-2'-deoxycytidine (decitabine) reduced the level of PDT-induced necrosis of glial cells but not neurons by 1.3 and 2.0 times, respectively, and did not significantly influence apoptosis of glial cells. Histone deacetylase inhibitors valproic acid and trichostatin A inhibited PDT-induced both necrosis and apoptosis of satellite glial cells but not neurons by 1.6-2.7 times. Thus, in the crayfish stretch receptor DNA methylation and histone deacetylation are involved in epigenetic control of glial but not neuronal necrosis. Histone deacetylation also participates in glial apoptosis.

Topics: Action Potentials; Animals; Apoptosis; Astacoidea; Azacitidine; Decitabine; DNA Methylation; Enzyme Inhibitors; Epigenesis, Genetic; Histone Deacetylase Inhibitors; Hydroxamic Acids; In Vitro Techniques; Indoles; Lasers; Mechanoreceptors; Necrosis; Neuroglia; Organometallic Compounds; Photic Stimulation; Photosensitizing Agents; Valproic Acid

Neuroglial interaction is very important for functioning and survival of nerve and glial cells. In the present work, we studied the influence of the intense neuronal activity on survival of the isolated crayfish stretch receptor neuron and surrounding glial cells subjected to photodynamic treatment, which induces intense oxidative stress. In the experimental group, neurons were stimulated by multiple extensions of the receptor muscle for 1h so that the firing rate did not fall below 10-15 Hz, whereas in the control group, the receptor muscles were relaxed and neurons were silent. After stimulation, the preparations were photosensitized with alumophthalocyanine Photosens and irradiated by 670 nm laser diode. The isolated stretch receptors were stained with propidium iodide and Hoechst 33342, which reveal the nuclei of the necrotic and the apoptotic cells, respectively. The level of apoptosis of photosensitized glial cells was significantly lower in the experimental group compared to the resting control. Necrosis of neurons and glial cells was not significantly influenced. Therefore, elevated neuronal activity increased the resistance of the surrounding glial cells to photoinduced apoptosis. This could be attributed to the depletion of the energetic resources, which are transferred from glia into the neuron to support its firing, or to the neurotrophic neuron-to-glia signaling.

Topics: Action Potentials; Animals; Apoptosis; Astacoidea; Cell Survival; In Vitro Techniques; Indoles; Lasers; Mechanoreceptors; Necrosis; Neuroglia; Organometallic Compounds; Photosensitizing Agents

The effects of Photodynamic Therapy using 2nd generation photosensitizers have been widely investigated aiming clinical application treatment of solid neoplasms. In this work, ultrastructure changes caused by the action of two 2nd generation photosensitizers and laser irradiation on CHO-K1 and HeLa (neoplastic) cells were analyzed by transmission electron microscopy. Aluminum phthalocyanine chloride, aluminum phthalocyanine tetrasulfonate chloride and radiation from a semiconductor laser at a fluency of 0.5 J/cm2 (Power=26 mW; lambda=.670 nm) were used. The results showed induction of apoptosis. Such alterations where observed in HeLa but not in CHO-K1 cells after Aluminum phthalocyanine tetrasulfonate chloride (AlPcS4, photodynamic treatment. The Aluminum phthalocyanine chloride (AlPc) photodynamic treatment induced necrosis on the neoplastic cell line, and cytoplasm and nuclear alterations on the normal cell line.

Topics: Animals; Apoptosis; Cell Nucleus; CHO Cells; Cricetinae; Cytoplasm; Female; HeLa Cells; Humans; Indoles; Lasers; Light; Microscopy, Electron; Mitochondria; Necrosis; Organometallic Compounds; Ovary; Photic Stimulation; Photochemotherapy; Radiation-Sensitizing Agents; Uterine Cervical Neoplasms

Topics: Animals; Cat Diseases; Cats; Disseminated Intravascular Coagulation; Indoles; Liver; Male; Necrosis; Organometallic Compounds; Photosensitizing Agents

The effects of photodynamic therapy (PDT) on normal brain tissue and depth of brain necrosis were evaluated in rats receiving 2.5 mg/kg aluminum phthalocyanine tetrasulfonate. Twenty-four hours later brains were irradiated with 675 nm light at a power density of 50 mW/cm2 and energy doses ranging from 1.6 to 121.5 J/cm2. Brains were removed 24 h after PDT and evaluated microscopically. When present, brain lesions consisted of well-demarcated areas of coagulation necrosis. When plotting the depth of necrosis against the natural log of energy dose, the data fit a piecewise linear model, with a changepoint at 54.6 J/cm2 and an x intercept of 7.85 J/cm2. The slopes before and after the changepoint were 2.04 and 0.21 mm/ln J cm-2, respectively. The x intercept suggests a minimum light dose below which necrosis of normal brain will not occur, whereas the changepoint indicates the energy density corresponding to an approximate maximum depth of necrosis.

Topics: Animals; Brain; Edema; Indoles; Light; Male; Necrosis; Organometallic Compounds; Photochemotherapy; Radiation-Sensitizing Agents; Rats; Rats, Inbred F344

We have previously reported photodynamic therapy of normal rat colon using aluminium sulphonated phthalocyanine (AISPc). In that study, the AISPc used was a mixture of phthalocyanines of different degrees of sulphonation. Phthalocyanines of defined degrees of sulphonation have recently become available and we compared the distribution of the di- and tetra-sulphonates (AIS2Pc and AIS4Pc) in rat colon and colon wall structures employing both chemical extraction and fluorescence photometry using a charge coupled device imaging system. Also, the photodynamic effects produced by these components in rat colon were compared at various times after photosensitization. After intravenous photosensitizer administration using equimolar doses, the concentration of AIS2Pc in colon fell off more rapidly with time than AIS4Pc. Differences were noted in the microscopic distribution of these compounds, with the di-sulphonate exhibiting peak fluorescence in colon wall structures by 1 h after photosensitization, while mucosal fluorescence with the tetra-sulphonate peaked at 5 h. Fluorescence was also lost from the colon wall much more slowly with the tetra-sulphonate, which tended to be retained in the submucosa. Maximum photosensitizing capability was seen at 1 h with AIS2Pc and no lesions could be produced with photodynamic therapy at 1 week, with up to 5.65 mumol/kg. With AIS4Pc (5.65 mumol/kg), while no lesions could be produced with light treatment at 1 h, photodynamic therapy at 1 week produced lesions only slightly smaller than those produced with treatment at 48 h (the time of maximum effect), and significant photosensitization was present at 2 weeks.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Aluminum; Animals; Colon; Female; Indoles; Necrosis; Organometallic Compounds; Photochemotherapy; Radiation-Sensitizing Agents; Rats; Rats, Inbred Strains