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Objective To observe the effects of acupuncture on TLR4, inflammation factors IL-1β and IL-6 expressions in the prefrontal cortex of vascular dementia (VD) rats; To investigate the brain protective mechanisms of acupuncture. Methods A total of 50 rats were randomly divided into sham-operation group, model group, acupuncture group, and placebo-acupuncture group. The animal model of VD was replicated by permanent bilateral common carotid artery occlusion (2VO) in rats. Acupuncture was performed at three days after surgery, once daily for two weeks. Morris water maze was used to test the cognitive function. The expressions of Iba1 and TLR4 were assessed by immunohistochemical method. Inflammation factors IL-1β and IL-6 mRNA expressions were tested by RT-PCR. Results Compared with sham-operation group, the model group rats showed impaired spatial learning and memory ability, and expression of Iba1, TLR4, and IL-1β, IL-6 increased. Compared with the model group, the spatial learning and memory abilities of the acupuncture group were improved, and the expression of Iba1, TLR4 and IL-1β, IL-6 in the prefrontal cortex decreased. These effects were not found in the placebo-acupuncture group. Conclusion The effect of acupuncture may be achieved by inhibiting microglia activate, decreasing the expressions of TLR4, IL-1β and IL-6 in the prefrontal cortex, resulting in improving spatial learning and memory ability of VD rats.
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Objective To observe the effects of acupuncture on the level of inflammatory cytokines and toll-like receptor 4 in hypothalamic paraventricular nucleus (PVN) of SHR; To investigate the mechanisms of acupuncture in reduction of mean arterial blood pressure (MAP) in SHR. Methods Thirty 10-week old SHR were randomly divided into SHR group, acupuncture group and non-acupoint group, with 10 rats in each group. 10 WKY rats were set as control group. Acupuncture group received bilateral acupuncture in "Taichong" acupoint, and twisting and diarrhea method was used to stimulate; non-acupoint group received acupuncture at the back of feet, and soothing and diarrhea with twisting method was used to stimulate. Materials were taken two weeks later. The mean arterial pressure of rats was detected every day; the expression of TLR4 mRNA in PVN was detected by RT-PCR; The expression of TLR4 protein in PVN was detected by Western blot; The levels of TNF-αa nd IL-6 were detected. Results Compared with control group, the mean arterial pressure of SHR group increased; TLR4 mRNA and protein expression in PVN increased significantly; the level of TNF-α and IL-6 increased significantly (P<0.05). Compared with SHR group, the mean arterial pressure of acupuncture group decreased significantly;TLR4 mRNA and protein expression in PVN decreased significantly; the level of TNF-α and IL-6 decreased significantly (P<0.05). There was no significant change in non-acupoint group. Conclusion Acupuncture in"Taichong" acupoint can attenuate blood pressure of SHR by inhibiting expression of TLR4 in PVN and reducing the levels of TNF-α and IL-6.
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Objective In order to fully reconstruct the accident by utilizing pedestrian injuries information gained from the car-pedestrian collision, a new method based on finite element simulation and genetic neural network to deduce the car-pedestrian collision parameters in reverse is proposed. Methods Crash simulations from different contact angles (back, left, front, right) at different impact speeds (25, 40, 55 km/h) were conducted by using Hyperworks and LS-DYNA, so as to obtain the head injury criterion (HIC) value and the maximum velocity of the thoracic wall. According to the criteria of injury biomechanics, the severities of the pedestrian head and thorax and corresponding injury locations were analyzed and set as predictors, and the predictive values of collision parameters were then acquired by using genetic neural network. Finally, this method was verified by two car-pedestrian accidents with the video and exact collision parameters. Results For both cases of the car-pedestrian accidents, the car speeds at the collision of pedestrian were 54 and 49 km/h, respectively, and the car-pedestrian contact angles were both 180°. While according to the pedestrian injuries information, the predictive values of the car speeds at the collision of pedestrian were 51 and 43 km/h, and the predictive values of the car-pedestrian contact angles were 184° and 169°, respectively. The reconstruction accuracies of two cases were 0.94 and 0.88. Conclusions The proposed method in the study can be used to predict car-pedestrian collision parameters efficiently and accurately by utilizing the pedestrian injuries information, which provides a new method for cause analysis and responsibility recognition, as well as theoretical references for the treatment and protection of head and thoracic injuries occurred in the car-pedestrian collision.
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<p><b>OBJECTIVE</b>To explore the effects of safflower injection on prevention and treatment of hypoxic pulmonary hypertension and clarify the function of the endoplasmic reticulum stress apoptosis pathway during the process.</p><p><b>METHODS</b>Thirty male SD rats were randomly grouped as normal control group, hypoxia-hypercapnia group and hypoxia+safflower group. The latter two groups were put in the cabin with oxygen concentration ranged from 9% to 11% and carbon dioxide concentration from 5% to 6%. The pulmonary artery pressure and the index of right ventricular hypertrophy were determined after hypoxia exposure (8 h/dx28 d). Changes in morphology of lung tissue were observed by electron microscopy. To explore the possible mechanisms, we also detected apoptosis and apoptosis-related genes/proteins in lung tissue by TUNEL reactivity and PCR and Western blot.</p><p><b>RESULTS</b>Compared with the normal control group, pulmonary artery pressure and the index of right ventricular hypertrophy in hypoxia group were 45% and 33.4% higher, respectively. Tiny blood vessel wall of lungs was thickened and edema, and proliferation of collagen fibers was obvious under the electron microscope. TUNEL staining of apoptotic cells in lung tissues showed more high brightness green fluorescence (+-++), but less green fluorescence showed in the pulmonary vascular smooth muscle cell layer, and apoptosis index (AI) value was 150% higher; gene and protein expression levels of endoplasmic reticulum stress pathway were increased. Compared with hypoxia-hypercapnia group, pulmonary artery pressure and the index of right ventricular hypertrophy in the hypoxia+safflower group were 18% and 15.6% lower, respectively; collagen fibers were decreased, and smooth muscle cells and epithelial cells were got apoptotic-like changes under the electron microscope. TUNEL staining of apoptotic cells in lung tissues showed brighter green fluorescence (++-+++); the high brightness green fluorescence showed in pulmonary vascular smooth muscle cell layer, and apoptotic index (Al) value was 40% higher; gene and protein expressions of endoplasmic reticulum stress pathway were significantly upregulated.</p><p><b>CONCLUSION</b>Our findings demonstrate that safflower injection could activate endoplasmic reticulum stress-induced apoptosis and especially promote apoptosis in pulmonary vascular smooth muscle cells.</p>
Subject(s)
Animals , Male , Rats , Apoptosis , Carthamus tinctorius , Chemistry , Endoplasmic Reticulum Stress , Hypercapnia , Hypertension, Pulmonary , Drug Therapy , Hypoxia , Lung , Cell Biology , Myocytes, Smooth Muscle , Rats, Sprague-DawleyABSTRACT
<p><b>OBJECTIVE</b>To investigate the changes of endoplasmic reticulum stress-induced apoptosis in pulmonary tissue of rats with hypoxic pulmonary hypertension.</p><p><b>METHODS</b>Twenty two male SD rats were randomly divided into control group and 4-week hypoxia-hypercapnia group (n=11). The mean pulmonary arterial pressure (mPAP) and the mean carotid arterial pressure (mCAP) were monitored, and the weight ratio of right ventricle (RV) to left ventricle plus septum (LV + S) were measured. The rattish pathological model were assessed by mPAP, mCAP, RV/(LV+ S), vessel wall area/total area (WA/TA), vessel cavity area/total area (CA/TA) and media thickness of pulmonary arteriole (PAMT). The pulmonary apoptotic cells were detected by Hoechst staining. RT-PCR was used to study the genetic expression of caspasel2, glucose regulated protein 78 (GRP78) and GRP94 in pulmonary tissue. The expression of GRP94 and GRP78 proteins in pulmonary tissue were determined by using immunohistochemistry.</p><p><b>RESULTS</b>(1) (The mPAP, RV/(LV + S), WA/TA and PAMT were respectively higher by 50.5%, 37.3%, 72.5% and 137% in hypoxic group than those in control group, while CA/TA was lower by 41.9% (all P < 0.01). There was not significant difference of mCAP between the two groups. (2) Hoechst staining showed that the pulmonary apoptotic cells in hypoxic group outnumbered markedly than those in control group, and the apoptotic cells were mainly in pulmonary tissue, while they were rare in pulmonary vascular smooth muscle cell. (3) Compared with control group, the expression of pulmonary caspasel2, GRP78 and GRP94 mRNA in hypoxic group were higher by 144%, 137% and 80.7% (all P < 0.05), respectively. (4) The expression of pulmonary GRP78 and GRP94 proteins were up-regulated in hypoxic group, and these proteins mainly localized in pulmonary vascular endothelial cell.</p><p><b>CONCLUSION</b>The endoplasmic reticulum stress-induced apoptosis may be one of the mechanism of hypoxic pulmonary hypertension and pulmonary vascular wall remodeling.</p>