RESUMO
OBJECTIVE: To observe the effect of moxibustion at Governor Vessel acupoints on inositol requiring enzyme 1 ï¼IRE1ï¼ / X-box binding protein 1 ï¼XBP1ï¼ pathway in hippocampal CA1 region of rats with vascular dementia ï¼VDï¼, so as to explore its mechanisms in the treatment of VD. METHODS: Male SD rats were randomly divided into normal, sham operation, model, moxibustion ï¼Moxiï¼ and medication groups ï¼n=12ï¼. The VD model was established by permanent ligation of bilateral common carotid arteries. For rats of the Moxi group, mild moxibustion was given to "Baihui" ï¼GV20ï¼, "Dazhui" ï¼GV14ï¼ and "Fengfu" ï¼GV16ï¼ for 20 min each point, once a day for consecutive 6 days per week, for a total of 4 weeks. For rats of the medication group, intragastric perfusion of nimodipine was given 3 times each day with total dose of 2 mgâ¢kg-1â¢d-1 for 4 weeks. Morris water maze test was used to detect the learning and memory ability of rats before and after modeling as well as after intervention. The apoptosis rate of nerve cells in hippocampal CA1 region was detected by TUNEL staining. The proteins and mRNA expression levels of IRE1, XBP1, B-cell lymphoma/leukemia-2 ï¼Bcl-2ï¼, Bcl-2 associated X protein ï¼Baxï¼ in hippocampal CA1 region were detected by Western blot and real-time quantitative PCR, respectively. RESULTS: Compared with the sham operation group, the average escape latency was significantly prolonged ï¼P<0.01ï¼, the number of times crossing the original platform was significantly reduced ï¼P<0.01ï¼, the apoptosis rate of nerve cells in hippocampal CA1 region was significantly increased ï¼P<0.01ï¼, the proteins and mRNA expression levels of IRE1, XBP1 and Bax were significantly increased ï¼P<0.01ï¼, and the expression levels of Bcl-2 protein and mRNA were significantly decreased ï¼P<0.01ï¼ in rats of the model group. After treatment, compared with the model group, the average escape latency was significantly shortened ï¼P<0.01ï¼, the number of times crossing the original platform was increased ï¼P<0.05ï¼, the apoptosis rate of nerve cells in hippocampal CA1 region was significantly decreased ï¼P<0.01ï¼, the protein and mRNA expression levels of IRE1, XBP1 and Bax were decreased ï¼P<0.05, P<0.01ï¼, and the expression levels of Bcl-2 protein and mRNA were increased ï¼P<0.05, P<0.01ï¼ in rats of the Moxi group and medication group. There was no significant difference in the above indexes between the Moxi group and the medication group. CONCLUSION: Moxibustion at the acupoints of Governor Vessel can improve the cognitive function of VD rats, and its mechanism may be related to regulating IRE1/XBP1 pathway, inhibiting the release of pro-apoptotic protein Bax, increasing the expression of anti-apoptotic protein Bcl-2, and thus inhibiting the apoptosis of hippocampal nerve cells.
Assuntos
Demência Vascular , Moxibustão , Masculino , Animais , Ratos , Ratos Sprague-Dawley , Região CA1 Hipocampal , Proteína X Associada a bcl-2/genética , Proteína 1 de Ligação a X-Box , Demência Vascular/genética , Demência Vascular/terapia , Proteínas Proto-Oncogênicas c-bcl-2 , InositolRESUMO
OBJECTIVES: To observe the effect of electroacupuncture (EA) on microglia (MG), Janus kinase-2 (JAK2) and signal transducer and activator of transcription-3 (STAT3) in hippocampal CA1 region of Alzheimer's di-sease (AD) rats, so as to explore its mechanisms in the treatment of AD. METHODS: Thirty-six male SD rats were randomly divided into sham operation, model and EA groups, with 12 rats in each group. The AD rat model was established by intraperitoneal injection of D-galactose combined with intrahippocampal injection of aggregated Aß25-35. The rats in the EA group were given EA (2 Hz/20 Hz, 2 mA) at "Baihui"(GV20) and"Shenting"(GV24) for 30 min, once daily, 6 days a week for 4 weeks. Morris water maze test was used to detect the learning and memory ability and spatial exploration ability of rats. HE staining was used to observe the pathological changes of hippocampus. The ultrastructure of hippocampal neurons was observed by transmission electron microscopy. The positive expression of MG marker io-nized calcium adaptor protein (Iba-1) in hippocampus was observed by immunofluorescence staining. The expression levels of serum inflammatory factor interferon-γ (IFN-γ) and transforming growth factor beta 1 (TGF-ß1) were detected by ELISA. The mRNA expression levels of JAK2, STAT3, inducible nitric oxide synthase (iNOS) and arginase-1 (Arg-1) in hippocampal CA1 region were detected by real-time quantitative PCR. The protein and phosphorylation levels of JAK2 and STAT3 in hippocampal CA1 region were detected by Western blot. RESULTS: Compared with the sham operation group, the escape latency of the model group was significantly prolonged (P<0.01), and the number of crossing the original platform was significantly reduced (P<0.01), the positive expression of Iba-1 in CA1 region, the content of serum IFN-γ, the relative mRNA expressions of JAK2, STAT3 and iNOS, and the protein and phosphorylation levels of JAK2 and STAT3 were significantly increased (P<0.01), while the content of serum TGF-ß1 and the relative expression of Arg-1 mRNA were significantly decreased (P<0.01). Compared with the model group, the escape latency of rats in the EA group was significantly shortened (P<0.01), the number of crossing the original platform was significantly increased (P<0.01), the positive expression of Iba1, the content of serum IFN-γ, the mRNA expressions of JAK2, STAT3 and iNOS, and the protein and phosphorylation levels of JAK2 and STAT3 were significantly decreased (P<0.05, P<0.01), while the content of serum TGF-ß1 and the expression of Arg-1 mRNA were significantly increased (P<0.01). Moreover, pathological and ultrastructural observation showed a reduction in the number of hippocampal neurons, changement of nuclear morphology, dilation of intercellular space, and decreased number of mitochondria in the model groupï¼these situations were relatively milder in the EA group. CONCLUSIONS: EA can improve the learning and memory function of AD rats, which may be associated with its functions in decreasing MG activities, and inhibiting the JAK2 / STAT3 signaling pathway in the hippocampus.
Assuntos
Doença de Alzheimer , Disfunção Cognitiva , Eletroacupuntura , Ratos , Masculino , Animais , Doença de Alzheimer/genética , Doença de Alzheimer/terapia , Microglia , Fator de Crescimento Transformador beta1/genética , Ratos Sprague-Dawley , Hipocampo , Disfunção Cognitiva/genética , Disfunção Cognitiva/terapia , RNA MensageiroRESUMO
The purpose of this preclinical study in a sheep model was to confirm the feasibility and safety of the LuX-Valve transjugular tricuspid valve (TV) replacement apparatus and to optimize the implantation procedure before beginning first-in-man study. The LuX-Valve was implanted in a sheep model (n = 8) via transjugular approach. Six of eight sheep underwent successful implantation procedure on beating heart. The first two sheep died during the prostheses deployment. In the remaining 6 sheep that survived, postoperative echocardiography results showed there was no paravalvular leakage (PVL) and central tricuspid regurgitation in 5 animals, whereas 1 animal had mild PVL. The mean transvalvular gradient was 1.1 ± 0.9 mm Hg at the 4-week follow-up. No right ventricular outflow tract (RVOT) obstruction, device malposition, pericardial effusion, coronary artery compression, or arrhythmias were observed. This technology may be a promising alternative for TR patients who are at high risk for open-heart surgery. Transjugular tricuspid valved-stent implantation. a Transjugular tricuspid valve replacement in a sheep model. b and c Valved stent. d, e, and f Schematic depiction of the implantation procedure.
Assuntos
Bioprótese , Implante de Prótese de Valva Cardíaca , Próteses Valvulares Cardíacas , Animais , Ovinos , Valva Tricúspide/diagnóstico por imagem , Ecocardiografia , Desenho de Prótese , Cateterismo Cardíaco , Resultado do TratamentoRESUMO
OBJECTIVE: To observe the effect of moxibustion preconditioning on learning-memory ability, Toll like receptor 4(TLR4)/nuclear factor-kappa B(NF-κB) signal pathway related proteins and microglia in rats with Alzheimer's disease (AD), so as to explore its possible mechanisms underlying improvement of AD. METHODS: Male SD rats were randomly divided into normal, sham operation, AD model and pre-moxibustion groups, with 9 rats in each group. Moxibustion was applied to "Baihui"(GV20), "Shenshu"(BL23) and "Zusanli"(ST36) for 15 min, once daily, 6 days as a course of treatment for 3 courses. At the end of moxibustion, the AD model was established by injection of Aß25-35 aggregation solution into the bilateral hippocampus. The sham operation group was only injected with the same amount of 0.9% Nacl solution. The spatial learning-memory ability of rats was detected by Morris water maze test, the ultrastructure of hippocampal neurons was observed by transmission electron microscope (TEM). The histopathological changes of hippocampus tissue were observed by HE staining, and the protein expression levels of TLR4 and NF-κB p65 in the hippocampus detected by Western blot, and the positive expressions of Iba-1, CD80 and CD206 in the hippocampal CA1 region were detected by immunofluorescence labeling. The contents of inflammatory factors IL-1ß, TNF-α and IL-10 in the hippocampus were measured by ELISA. RESULTS: Compared with the sham operation group, the escape latency was significantly increased (P<0.01), and the number of platform quadrant crossing times was decreased (P<0.01) in the model group. In comparison with the model group, the increased escape latency and the decreased platform quadrant crossing times were reversed in the pre-moxibustion group (P<0.01). TEM and light microscope observation showed loose arrangement of cells, enlarged cell space, degeneration, swelling and deformation of hippocampal neurons, rupture of membranes of a large number of cells, reduction of mitochondria, dilation of endoplasmic reticulum, and matrix vacuoles, uneven distribution of organelles and cytoplasm, and being difficult in distinguishing the nuclear cytoplasm in the model group, which was relatively milder in the pre-moxibustion group. The expression levels of hippocampal NF-κB p65 and TLR4, the mean immunofluorescence density of Iba-1 and CD80, as well as the contents of IL-1ß and TNF-α in hippocampal CA1 region were significantly increased in the model group than those in the sham operation group (P<0.01), and obviously decreased in the pre-moxibustion group than those in the model group (P<0.05, P<0.01). Whereas the expression of CD206 and the content of IL-10 were evidently decreased in the model group than those in the sham operation group (P<0.01), and strikingly increased in the pre-moxibustion group than those in the model group (P<0.01). No significant differences were found between the sham operation group and the normal group in all the indexes mention above (P>0.05). CONCLUSION: Pre-moxibustion at GV20, BL23 and ST36 can improve learning-memory ability in AD rats, which may be associated with its functions in promoting the polarization of microglia from M1 to M2 and reducing the neuroinflammatory response by way of TLR4/NF-κB signaling pathway.
Assuntos
Doença de Alzheimer , Moxibustão , Masculino , Animais , Ratos , Ratos Sprague-Dawley , NF-kappa B/genética , Interleucina-10 , Microglia , Doença de Alzheimer/genética , Doença de Alzheimer/terapia , Receptor 4 Toll-Like/genética , Fator de Necrose Tumoral alfa , Transdução de SinaisRESUMO
BACKGROUND: The Makorin ring finger protein 1 (MKRN1) gene, also called RNF61, is located on the long arm of chromosome 7 and is a member of the RING finger protein family. The E3 ubiquitin ligase MKRN1 is closely linked to tumour development, but the exact mechanism needs to be elucidated. In this study, we aimed to investigate the specific mechanism and role of MKRN1 in colorectal cancer (CRC) development. METHODS: MKRN1 expression in CRC was analysed using the Cancer Cell Line Encyclopaedia and the Cancer Genome Atlas (TCGA) databases. Rectal tumour tissues were frozen to explore the MKRN1 expression in CRC and its clinical significance. The impact of MKRN1 on CRC cell proliferation and migration was observed using CCK8, colony formation, wound healing, and transwell assays. A combination of MKRN1 quantitative proteomics, ubiquitination modification omics analysis, and a string of in vitro and in vivo experiments revealed the potential mechanisms by which MKRN1 regulates CRC metastasis. RESULTS: MKRN1 expression was significantly elevated in CRC tissues compared to paracancerous tissues and was positively linked with prognosis (P < 0.01). MKRN1 downregulation inhibits CRC cell proliferation, migration, and invasion. Conversely, MKRN1 overexpression promotes the proliferation, migration, and invasion of CRC cells. Mechanistically, MKRN1 induces epithelial-mesenchymal transition (EMT) in CRC cells via ubiquitination and degradation of Smad nuclear-interacting protein 1 (SNIP1). Furthermore, SNIP1 inhibits transforming growth factor-ß (TGF-ß) signalling, and MKRN1 promotes TGF-ß signalling by degrading SNIP1 to induce EMT in CRC cells. Finally, using conditional knockout mice, intestinal lesions and metastatic liver microlesions were greatly reduced in the intestinal knockout MKRN1 group compared to that in the control group. CONCLUSIONS: High MKRN1 levels promote TGF-ß signalling through ubiquitination and degradation of SNIP1, thereby facilitating CRC metastasis, and supporting MKRN1 as a CRC pro-cancer factor. The MKRN1/SNIP1/TGF-ß axis may be a potential therapeutic target in CRC.
Assuntos
Neoplasias Colorretais , Proteínas de Ligação a RNA , Ribonucleoproteínas , Animais , Camundongos , Linhagem Celular , Proliferação de Células , Neoplasias Colorretais/genética , Proteólise , Humanos , Ribonucleoproteínas/metabolismo , Proteínas de Ligação a RNA/genética , Fator de Crescimento Transformador beta/metabolismo , Transdução de SinaisRESUMO
OBJECTIVE: To investigate the effect of melatonin (MLT) on the proliferation and apoptosis of human multiple myeloma cell line RPMI 8226 and its possible mechanism. METHODS: RPMI 8226 cells were cultured in vitro, and different concentrations of MLT were treated on RPMI 8226 cells. The effects of MLT on RPMI 8226 cell proliferation were detected by CCK-8 assay and methylcellulose cloning assay, and the effects of MLT on cell apoptosis were detected by AnnexinV-FITC /PI, flow cytometry. Western blot was used to determine the expression of apoptosis and endoplasmic reticulum stress-related proteins in each group, and CCK-8 assay was used to determine the effect of MLT combined with bortezemib on the viability of RPMI 8226 cells. RESULTS: MLT inhibited the proliferation of RPMI 8226 cells in a dose- and time-dependent manner (r=-0.9777ï¼r=-0.9951). With the increase of MLT concentration, the number of clones decreased, the apoptosis of RPMI 8226 cells increased (P<0.05), the expression of anti-apoptotic protein XIAP decreased, the expression of apoptotic proteins Bax and Caspase3 increased, and the expression of endoplasmic reticulum stress-related proteins increased. Compared with the control group, the survival of RPMI 8226 cells in the MLT and BTZ combined group significantly decreased (P<0.01). CONCLUSION: MLT can inhibit the proliferation of RPMI 8226 cells, promote the apoptosis of RPMI 8226 cells, and enhance the anti-tumor effect of BTZ on RPMI 8226 cells. The mechanism may be related to endoplasmic reticulum stress.
Assuntos
Melatonina , Mieloma Múltiplo , Apoptose , Linhagem Celular Tumoral , Proliferação de Células , Estresse do Retículo Endoplasmático , Humanos , Melatonina/farmacologia , Mieloma Múltiplo/patologiaRESUMO
OBJECTIVE: To observe the effect of electroacupuncture (EA) on miRNA-126-3p and mammalian target of rapamycin (mTOR)/hypoxia-inducible factor-1α (HIF-1α) signaling pathway in rats with cerebral ischemia (CI), so as to explore the underlying mechanism of EA on angiogenesis. METHODS: Male SD rats were randomly divided into control group, model group, EA group and EA+inhibitor group (inhibitor group), which were further divided into 3, 7 and 14 d subgroups, with 12 rats in each sub-group. The CI model was established by occlusion of the middle cerebral artery. EA (2 Hz/20 Hz, 0.5 mA) was applied to "Dazhui" (GV14), "Baihui" (GV20) for 20 min, once daily for 14 days at most. Rats of the inhibitor group were given an intraperitoneally injection of mTOR inhibitor (0.1 mg/mL, 0.3 mg/kg) before daily EA. The neurological function was evaluated by modified neurological severity score (mNSS). The ultrastructure of cortical neurons and microvascular endothelial cells in ischemic penumbra was observed by transmission electron microscope, and the microvessel density (MVD) of cortical endothelium in ischemic penumbra was detected by immunohistochemistry. Western blot and quantitative real-time PCR were used to detect the protein and mRNA expression of mTOR, HIF-1α and the expression of miR-126-3p in the cortex of ischemic penumbra, respectively. RESULTS: After modeling, compared with the control group at the same time point, the mNSS of the model group was increased (P<0.01), and decreased over time (P<0.01). The cortical neurons and brain microvascular endothelial cells in the ischemic penumbra were edema, and the cell structure was damaged obviously in the model group.The MVD value and the expressions of mTORãHIF-1α proteins and mRNAs were increased (P<0.01), while the expression of miR-126-3p decreased (P<0.01) in the model group relative to the control group. Compared with the model group at the same time point, the mNSS of both intervention groups was significantly reduced (P<0.01, P<0.05), the neuron and cerebral microvascular structure improved to varying degrees, and the MVD value, the expressions of mTOR and HIF-1α protein and mRNA, and the expression of miR-126-3p of the two treatment groups were increased (P<0.01, P<0.05) at all time points (excep MVD at day 7 in the inhibitor group). Compared with the EA group at the same time point, MVD, the expressions of mTOR, HIF-1α proteins and mRNAs and miR-126-3p in the inhibitor group were all decreased (P<0.05,P<0.01). Compared with the group itself at 4 hours after modeling and day 3 and day 7, the mNSS was decreased at day 14 ï¼P<0.01ï¼ in the model, EA and inhibitor groups. Compared with the group itself at day 3, the MVD value and the expression of mTOR protein were increased at day 7 and day 14 in the model, EA and inhibitor groups (P<0.01, P<0.05). Compared with the group itself at day 3 and day 7, the expression of mTOR mRNA and miR-126-3p were up-regulated at day 14 in the model and EA groups (P<0.01, P<0.05).Compared with the group itself at day 3, the mRNA expressions of mTOR and HIF-1α were increased at day 7 and day 14 (P<0.01, P<0.05) in the inhibitor group. CONCLUSION: EA at GV14 and GV20 can alleviate neurological deficit and improve angiogenesis in rats with CI, which may be related with its effect in up-regulating the expression of mTOR and HIF-1α, improving activation of miR-126-3p in the cortex of ischemic penumbra.
Assuntos
Isquemia Encefálica , Eletroacupuntura , MicroRNAs , Animais , Isquemia Encefálica/genética , Isquemia Encefálica/terapia , Infarto Cerebral , Células Endoteliais , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Isquemia , Masculino , MicroRNAs/genética , Neovascularização Fisiológica , RNA Mensageiro , Ratos , Ratos Sprague-Dawley , Transdução de Sinais , Serina-Treonina Quinases TOR/genéticaRESUMO
OBJECTIVE: To observe the effect of pre-moxibustion at "Baihui"(GV20), "Shenshu"(BL23) and "Zusanli"(ST36) on expression of Tau protein and related protein kinases as glycogen synthase kinase-3ß (GSK-3ß), etc. in the hippocampal CA3 region of Alzheimer's disease (AD) rats, so as to explore its mechanism underlying prevention and treatment of AD cognitive impairment. METHODS: Male SD rats were randomly divided into 4 groups: normal control, sham operationï¼ model and pre-moxibustionï¼with 9 rats in each group. Rats of the pre-moxibustion group received moxibustion of GV20, BL23 and ST36 for 15 min, once a day, 6 days a week for 3 weeks. After completion of moxibustion, the AD model was reproduced by injection of amyloid beta-peptide 25-35(Aß 25-35) aggregation solution 1 µL (5 µg/µL) into the bilateral hippocampus, rats of the sham operation group received injection of the same dose of normal saline into the hippocampus. The spatial learning-memory ability was detected using Morris water maze test, and changes of the ultrastructure of hippocampal neurons were observed using electron microscope, and those of histopathological changes of hippocampus tissue observed using hematoxylin eosin (H.E.) staining. The expression levels of hippocampal GSK-3ß, p-Tau, CDK5 and Synapsin I proteins were detected by Western blot and immunohistochemistry, respectively. RESULTS: No significances were found between the normal control and sham groups in all the indexes (P>0.05). Compared with the control group, the escape latency of place navigation test of Morris water maze test, expression of GSK-3ß and CDK5 and the immunoactivity of GSK-3ß, CDK5 and p-Tau were significantly increased (P<0.01), and the residence time in the platform quadrant and the number of platform crossing of spatial prob test and the expression of Synapsin â significantly reduced in the model group (P<0.01). Following the intervention, the increase of escape latency, expression of GSK-3ß and CDK5 and the immunoactivity of GSK-3ß, CDK5 and p-Tau, and the decrease of residence time in the platform quadrant, number of platform crossing and the expression of Synapsin â were reversed in the pre-moxibustion group (P<0.05, P<0.01). Outcomes of ultrastructure and histopathological observations respectively showed edema of hippocampal nerve cells at varying degrees, moderate edema of the cytoplasma, chromatin condensation at the edge of the nucleus, partial mitochondrial vacuole-like degeneration, fracture of tubular crest, edema and expansion of Golgi body, disappearance of polarity, fracture of the rough endoplasmic reticulum, degeneration of ribosome and partial myelin axon and reduced synaptic vesicles in the presynaptic capsule; and reduced number of neurons with shrank body, disappearance of nucleolus and blurred nuclear boundary and vacuole-like degeneration in some of them in the model group, which were relatively milder in the pre-moxibustion group. CONCLUSION: Pre-moxibustion at GV20, BL23 and ST36 plays a role in slowing down the occurrence and development of cognitive impairment in AD rats, which may be related to its functions in inhibiting tau protein hyperphosphorylation and reducing the expression of some related protein kinases in the hippocampus.
Assuntos
Doença de Alzheimer , Moxibustão , Doença de Alzheimer/genética , Doença de Alzheimer/terapia , Peptídeos beta-Amiloides , Animais , Região CA3 Hipocampal , Glicogênio Sintase Quinase 3 beta/genética , Hipocampo , Masculino , Proteínas Quinases , Ratos , Ratos Sprague-Dawley , Sinapsinas , Proteínas tau/genéticaRESUMO
OBJECTIVE: To observe the protective effect of electroacupuncture (EA) on neurovascular unit, neurological function in rats with cerebral ischemia (CI), so as to explore its mechanisms underlying improvement of ischemic cerebral tissue. METHODS: Male SD rats, SPF grade, were randomly and equally divided into sham operation group, model group, EA group â and EA group â ¡ï¼27 rats in each group. The CI model was established by occlusion of the middle cerebral artery (MCAO). EA (2 Hz/20 Hz, 0.5 mA) was applied to "Quchi"(LI11), "Hegu"(LI4), "Zusanli"(ST36) and "Shuigou"(GV26) for rats of the EA group â , and to "Baihui"(GV26), "Fengfu"(GV16), "Neiguan"(PC26) and "Xinshu"(BL15) for rats of the EA group â ¡ for 20 min, once a day for 14 days. The modified neurologic severity score (mNSS) was calculated according to the state of locomotor, sensory, and reflex parameters. Transmission electron microscope (TEM) was used to observe the neuronal structure of the ischemic cerebral area. The CD34 positive cells (for microvessels) of the ischemic brain tissue were detected by using immunohistochemistry, and the expression levels of cerebral phosphatidylinositol-3 kinase (PI3K) and protein kinase B (Akt) mRNAs were detected by quantitative real-time-PCR, respectively. RESULTS: Along with the extension of time, the mNSS at 4 h, and 3, 7 and 14 d after CI were apparently decreased, and the number of CD34 positive cells from 3 d to 14 d after CI, and the expression of PI3K mRNA and Akt mRNA from 3 d to 7 d were significantly increased in the modelï¼EAâ and EA â ¡ group ï¼P<0.01, P<0.05ï¼. Compared with the sham operation group, the mNSS at 4 h, and 3, 7 and 14 d, and CD34-positive number and PI3K mRNA and Akt mRNA expression levels at 3, 7 and 14 d were significantly increased in the model group (P<0.01, P>0.05). In comparison with the model group, the mNSS at 3, 7 and 14 d were obviously decreased (P<0.01), and the CD34-positive number and PI3K and Akt mRNA expression levels at 3, 7 and 14 d considerably increased in both EA group â and â ¡ (P<0.01, P<0.05). The therapeutic effect of EA group â ¡ was significantly superior to that of EA group â in lowering mNSS at 14 d, up-regulating the CD34-positive number at 7 and 14 d,and PI3K mRNA at 3, 7 and 14 d and Akt mRNA at 3 and 7 d (P<0.05, P<0.01). Results of TEM showed an irregular shape of neurons with nuclear pyknosis, non-uniform chromatin, more organelle loss, swollen mitochondrial Golgi complex and expansion of rough endoplasmic reticulum, being relatively milder in the EA group â , particularly in the EA group â ¡. CONCLUSION: EA therapy can improve the neurological function in cerebral ischemia rats, which may be related to its effects in protecting the neurovascular unit and up-regulating PI3K/Akt signal pathway. The effects of EA at GV26, GV16, PC26 and BL15 are better than those of EA at LI11, LI4, ST36 and GV26.
Assuntos
Isquemia Encefálica , Eletroacupuntura , AVC Isquêmico , Traumatismo por Reperfusão , Acidente Vascular Cerebral , Pontos de Acupuntura , Animais , Isquemia Encefálica/genética , Isquemia Encefálica/terapia , Masculino , Fosfatidilinositol 3-Quinases/genética , Proteínas Proto-Oncogênicas c-akt/genética , Ratos , Ratos Sprague-Dawley , Transdução de Sinais , Acidente Vascular Cerebral/genética , Acidente Vascular Cerebral/terapiaRESUMO
OBJECTIVE: To observe the effect of electroacupuncture (EA) combined with rehabilitation training on regional cerebral blood flow (rCBF) and angiogenesis in rats with acute cerebral ischemia (ACI), so as to explore its mechanisms underlying improvement of ACI. METHODS: A total of 135 male SD rats were divided into 5 groups: sham-operation (sham), model, EA, rehabilitation training and EA+rehabilitation training (combined treatment) groups (n=27 rats in each group). The ACI model was established by occlusion of the middle cerebral artery with thread embolus. EA (2 Hz/20 Hz, 3-5 V) was applied to "Baihui" (GV20), "Shuigou" (GV26) and bilateral "Neiguan" (PC6) for 20 min, once daily for 14 days. The rehabilitation training including hair-brushing in an enriched environment (10 min), round wooden-stick turning (10 min), grid-board climbing (10 min), and treadmill running (30 min/d) was condacted once daily for 14 days. The rCBF was measured by Doppler ultrasound. The cerebral infarct volume (CIV) was measured after 2, 3, 5-triphenyltetrazolium chloride (TTC) staining. The expression of CD34+ in the ischemic penumbra region of brain tissue was detected by immunohistochemistry, and the expressions of angiogenesis-related factors as vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 2 (VEGFR2) and basic fibroblast growth factor (bFGF) proteins in the ischemic brain tissue were detected by Western blot. RESULTS: Following modeling, the rCBF levels at the 5 min, 3rd, 7th and 14th day were significantly decreased in the model group relevant to the sham group (P<0.01). After the intervention, the rCBF levels were significantly increased on day 3, 7 and 14 in the combined treatment group and on day 7 and 14 in both the EA and rehabilitation training groups in comparison with the model group (P<0.01). The CIV was obvious in the model group in comparison with the sham group (P<0.01), but was markedly smaller in the EA, rehabilitation training and combined treatment groups on day 3,7 and 14 than in the model group (P<0.01). The number of CD34+ positive cells, and the expression levels of VEGF, VEGFR2, and bFGF proteins in ischemic brain tissues were significantly higher on day 3, 7 and 14 in the model group than in the sham group (P<0.01, P<0.05), and were further up-regulated considerably at the 3 time-points in the 3 treatment groups (P<0.01, P<0.05). The therapeutic effect of EA+rehabilitation training was significantly superior to that of simple EA and simple rehabilitation training in up-regulating rCBF, CD34+ positive cell number, and expression levels of VEGF, VEGFR2 and bFGF, and in down-regulating the CIV on day 3,7 and 14 (P<0.05, P<0.01). No significant differences were found between the EA and rehabilitation groups in the above-mentioned 6 indexes (P>0.05). CONCLUSION: EA combined with rehabilitation training can reduce the infarct volume and increase rCBF in ACI rats, which is probably associated with its effects in promoting the expression of angiogenesis-related factors of ischemic brain tissues. The effect of EA combined with rehabilitation training is markedly better than that of EA and rehabilitation training alone.