RESUMEN
OBJECTIVE: To observe the effect of acupuncture on activities of microglia in traumatic brain injury (TBI) rats. METHODS: Fifty-four male SD rats were randomly and equally divided into normal control, model and acupuncture groups according to the random number table (nï¼18 rats in each group). The TBI model was established by using a free fall brain injury striking device after exposing the local cranial bone (to induce the left parietal cerebral contusion). Acupoints "Baihui" (GV20), "Shuigou" (GV26), "Fengfu" (GV16), "Yamen" (GV15) and bilateral "Hegu" (LII4) were stimulated intensively by twirling the filiform needles with force at a range of >360° and a frequency of 160ï¼180 cycles/min for 10 sec in every acupoint, once every 5 min during the 15 minutes' needle retaining. The treatment was given once every day for successive 14 days. The rats of the normal and model groups were grabbed and fixed with the same procedure. The behavioral changes were tested using modified neurological severity score (mNSS). The histopathological changes of the injured cerebral cortex tissues were observed by using hematoxylin-eosin (H.E.) staining, and the fluorescence intensity of Iba-1 (marker of microglia) positive products in the surrounding tissue of the cerebral focus was displayed by immunofluorescence staining, and the contents of neuron specific enolate (NSE) and neurite outgrowth inhibitor-A (Nogo-A) in serum (indicating a secondary nerve damage) were assayed by ELISA. RESULTS: The mNSS scores were significantly increased on day 1, 3, 7 and 14 in the model group in comparison with the normal group (P<0.01) and considerably decreased at the 4 time-points after acupuncture intervention relevant to the model group (P<0.05, P<0.01). Hï¼E. staining showed that modeling induced pathological changes such as the excursion of cell nucleus, cellular swel-ling, vacuole-like change, neuron death, karyopyknosis dissolution, and proliferation of fibrous tissue were relatively milder in the acupuncture group. The average fluorescence intensity values of Iba-1-positive products, serum NSE and Nogo-A contents on day 3, 7 and 14 were significantly higher in the model group than in the normal group (P<0.05, P<0.01), and notably down-regulated in the acupuncture group than in the model group (P<0.05, P<0.01, except Nogo-A on day 3). CONCLUSION: Acupuncture intervention may accelerate neurological function recovery in TBI rats, which is closely related to its effects in inhibiting the activation of microglia and secondary nerve damage.
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Terapia por Acupuntura , Lesiones Traumáticas del Encéfalo , Animales , Masculino , Microglía , Proteínas Nogo , Ratas , Ratas Sprague-DawleyRESUMEN
OBJECTIVE: To study the effect of acupuncture on the TLR2/4-NF-κB signalling pathway in the cortex of Sprague-Dawley rats following traumatic brain injury (TBI), and investigate the possible mechanism underlying the effects of acupuncture on scar repair. METHODS: TBI was established using Feeney's free-falling epidural percussion model. In total, 108 rats were randomly divided into a normal group (n=18), untreated TBI model group (TBI group, n=36) and manual acupuncture-treated TBI group (TBI+MA, n=36). Each group of rats was subdivided into three time groups: 3-day (3d), 7-day (7d) and 14-day (14d). No treatment was given to rats in the normal and TBI groups. The TBI+MA group received manual acupuncture at GV20, GV26, GV16 through GV15, and bilateral LI4. mRNA expression of TLR2, TLR4, NF-κB and protein in the rat cortices was quantified using real-time fluorescence quantitative polymerase chain reaction (qPCR) and Western blot analyses. RESULTS: The modified neurological severity score (mNSS) scores of the TBI+MA group were improved compared with baseline scores 12 hours after modelling, and improved at 7d and 14d compared with the TBI group (P<0.05), while the score of the TBI group did not improve until 14d compared to baseline. mRNA and protein expression of TLR2, TLR4 and NF-κB in the TBI group were higher than the normal group at 3d (P<0.05), reached a peak at 7d, then began to decrease at 14d. mRNA and protein expression of TLR2, TLR4 and NF-κB were higher in the TBI+MA group compared with the TBI group at 3d (P<0.05), were significantly down-regulated at 7d (P<0.01), and decreased to normal levels at 14d. CONCLUSIONS: Acupuncture has a bidirectional regulatory effect on the TLR2/4-NF-κB signalling pathway-related genes TLR2, TLR4 and NF-κB in the TBI rat cortex, promoting their expression in the early stage and inhibiting it in the later stage.
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Terapia por Acupuntura , Lesiones Traumáticas del Encéfalo/terapia , FN-kappa B/metabolismo , Receptor Toll-Like 2/metabolismo , Puntos de Acupuntura , Animales , Lesiones Traumáticas del Encéfalo/genética , Lesiones Traumáticas del Encéfalo/metabolismo , Humanos , Masculino , FN-kappa B/genética , Ratas , Ratas Sprague-Dawley , Transducción de Señal , Receptor Toll-Like 2/genéticaRESUMEN
OBJECTIVE: To explore a new method of establishing HepG2 cell model of steatosis and observe the expression and significance of nuclear factor erythroid-2p45-related factor 2(Nrf2)/antioxidative response element (ARE) pathway related factors in HepG2 cells of steatosis. METHODS: HepG2 cells were induced with DMEM containing 25% fetal bovine serum, 0.1% MCT/LCT Fat Emulsion and 0.1 mmol/L free fatty acid (FFA) at different stages and the control group cells were cultured with normal DMEM medium. After the cell models were successfully established, lipid droplets in cytoplasm were observed with Oil Red 0 staining, and the triglyceride (TG) accumulation in HepG2 cells were tested by biochemical assay. Intracellular reactive oxygen species (ROS) concentration were detected by flow cytometry. Nitric oxide (NO), superoxide dismutase(SOD), malonyldialdehyde(MDA) and glutathione peroxidase(GSH-Px) were tested by biological reagent kit, while the protein expression of nuclear factor erythroid-2p45-related factor 2(Nrf2), heme oxygenase-1 (HO-1) and NAD(P)H: quinone oxidoreductase-1(NQO1) were analyzed by Western blot. RESULTS: Compared with that in the control group, red cytoplasmic lipid droplets were visible in model group; TG,ROS, NO, MDA concentration (P < 0.05, P < 0.01) and the protein expression of Nrf2, HO-1 and NQO1 (P < 0.05, P < 0.01)were significantly higher in model group, while SOD, GSH-Px concentration reduced significantly (P < 0.01). CONCLUSION: The in vitro cell model of steatosis and oxidative stress was successfully established. The activation of Nrf2/ARE pathway related factors maybe relevant to the overreaction of oxidative stress in HepG2 cells of steatosis.
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Elementos de Respuesta Antioxidante , Hígado Graso , Factor 2 Relacionado con NF-E2 , Estrés Oxidativo , Humanos , Medios de Cultivo , Ácidos Grasos no Esterificados , Hígado Graso/metabolismo , Glutatión Peroxidasa/metabolismo , Hemo-Oxigenasa 1/metabolismo , Células Hep G2 , Malondialdehído/metabolismo , NAD(P)H Deshidrogenasa (Quinona)/metabolismo , Factor 2 Relacionado con NF-E2/metabolismo , Óxido Nítrico/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Superóxido Dismutasa/metabolismo , Triglicéridos/metabolismoRESUMEN
OBJECTIVE: To observe the effects of acupuncture treatment on the expression of Wnt/ß-catenin signalling pathway-related genes (Wnt3a, ß-catenin and Sox2) in the injured cerebral cortex of rats with traumatic brain injury (TBI). METHODS: A controlled impact model of TBI was established using Feeney's free-drop method. Seventy-eight Sprague-Dawley rats were randomly divided into the following three groups: a normal group (n=18) that was left untreated; a model group (n=30) that received no treatment after TBI; and an acupuncture group (n=30) that received acupuncture (at LI4, GV20, GV26 and GV16) after TBI. Rats in each group were randomly and equally divided into 3-day, 7-day and 14-day subgroups according to the duration of therapy. Real-time fluorescence quantitative PCR (RT-qPCR) was used to measure mRNA expression of Wnt3a, ß-catenin and Sox2. Western blots were performed to determine the expression levels of WNT3a, ß-Catenin and SOX2. RESULTS: Wnt3a mRNA was upregulated in the 7-day and 14-day acupuncture subgroups compared with the corresponding model subgroups (p<0.05). ß-catenin expression was significantly increased in the 7-day and 14-day acupuncture subgroups compared with the corresponding model subgroups (p<0.01). In the 3-day and 7-day acupuncture subgroups, Sox2 expression was significantly higher than that in the normal and model groups (p<0.01 each). The levels of WNT3a, ß-catenin and SOX2 were generally consistent with the corresponding mRNA levels. CONCLUSIONS: Acupuncture exerts a regulatory effect on the Wnt/ß-catenin signalling pathway, which may in turn influence the proliferation and differentiation of endogenous neural stem cells.
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Terapia por Acupuntura , Lesiones Encefálicas/terapia , Factores de Transcripción SOXB1/metabolismo , Vía de Señalización Wnt , Proteína Wnt3A/metabolismo , beta Catenina/metabolismo , Animales , Lesiones Encefálicas/genética , Lesiones Encefálicas/metabolismo , Humanos , Masculino , Ratas , Ratas Sprague-Dawley , Factores de Transcripción SOXB1/genética , Proteína Wnt3A/genética , beta Catenina/genéticaRESUMEN
OBJECTIVE: To observe the effect of acupuncture on proliferation and differentiation of neural stem cells in brain tissues of rats with traumatic brain injuny. METHODS: Thirty SD rats were randomly and equally allocated to the sham-operated, the model and the acupuncture groups. The traumatic brain injury model was established by the free drop method. For the rats in the acupuncture group, acupuncture was applied once a day for 7 days. Brain histotomy was carried out when treatments were completed. Immunohistochemical techniques were adopted to detect the cells that express nestin, neurofilament proteins (NF)-200 and glial fibrillary acidic proteins (GFAP), the markers of neural stem cells, neurons, astrocytes respectively. RESULTS: Compared to the sham-operated group, the number of nestin-positive cells and NF-200-positive cells in brain tissues was decreased significantly in the model group (P < 0.01), whereas the number of GFAP-positive cells was significantly increased P<0.01). Compared to the model group, the positive cells of nestin, NF-200, GFAP in brain tissues in the acupuncture group were increased obviously (P<0.01). CONCLUSIONS: Acupuncture can significantly increase the number of nestin-positive cells, NF-200-positive cells and GFAP-positive cells, indicating the significant increase of neural stem cells, neurons and astrocytes in number. Acupuncture can improve neuranagenesis by promoting the proliferation and differentiation of neural stem cells in brain tissues. This might be one of the mechanisms for acupuncture to treat traumatic brain injury and to promote the repair of nervous function.
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Terapia por Acupuntura , Lesiones Encefálicas/patología , Lesiones Encefálicas/terapia , Encéfalo/patología , Diferenciación Celular , Células-Madre Neurales/patología , Animales , Proliferación Celular , Corteza Cerebral/patología , Proteína Ácida Fibrilar de la Glía/metabolismo , Proteínas de Filamentos Intermediarios/metabolismo , Masculino , Proteínas del Tejido Nervioso/metabolismo , Nestina , Células-Madre Neurales/metabolismo , Proteínas de Neurofilamentos/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
Primary splenic diffuse large B-cell lymphoma (DLBCL) is a rare clinical condition, which is generally treated by six to eight cycles of chemotherapy involving a combination of rituximab and the cyclophosphamide, adriamycin, vincristine, and prednisolone (CHOP) regimen. However, the treatment for chemorefractory primary splenic DLBCL remains controversial. Therapeutic splenic irradiation (SI) might be a reasonable and possibly the only treatment option with curative intention for patients with chemorefractory primary splenic DLBCL. However, the efficacy and safety of therapeutic SI are unclear. Herein, we present the case of a primary splenic DLBCL patient who was refractory to multiple chemotherapy regimens but achieved complete remission after administration of therapeutic SI. However, his condition was complicated with severe gastric variceal bleeding due to splenic venous thrombosis, which was successfully treated via splenectomy and short gastric vein ligation. On the basis of our findings, we concluded that the splenic venous thrombosis-induced gastric variceal bleeding was a rare but life-threatening adverse effect of the therapeutic SI administered for primary splenic DLBCL. Surgical intervention involving splenectomy and short gastric vein ligation is mandatory and should be performed as soon as possible for such patients.