共查询到10条相似文献,搜索用时 62 毫秒
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探讨流式细胞术在检测高糖诱导的早期雪旺细胞凋亡中应用。选用培养大鼠的雪旺细胞,将细胞分为正常对照组(5.6mmol/L葡萄糖),高糖组(50mmol/L葡萄糖),高渗出组(50mmg/L甘露醇),培养7天后,收集细胞。48h后应用流式细胞术采用Annexin-V/PI检测法和JC-1检测早期细胞的细胞膜凋亡率和线粒体膜电位变化。结果表明:高糖与正常糖浓度处理的雪旺细胞比较,高浓度葡萄糖诱导雪旺凋亡率明显高于正常葡萄组的凋亡率,线粒体膜电位明显高于正常对照组。高渗透组与正常对照组凋亡率及线粒体无明显升高。通过采用流式细胞细胞术结合单克隆抗体,能够快速、灵敏和准确检测雪旺细胞在细胞膜的通透性改变和线粒体膜电位变化。为研究雪旺细胞凋亡提供客观、直接、准确的方法。 相似文献
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流式细胞术快速检测T细胞亚群和NKT细胞亚群的免疫荧光变化 总被引:1,自引:0,他引:1
利用单克隆抗体与免疫荧光结合的特异性,探讨流式细胞仪检测T细胞亚群和NKT细胞亚群免疫荧光变化。选用培养C57BL/7小鼠脾细胞分别经SEB.COA活化,收集体外扩增10d淋巴细胞为效应细胞,经免疫荧光探针FITC、PI、PerCP和APC标记可同时检测各亚群免疫荧光变化,结果表明SEB活化主要是NKT细胞,它们与COA活化NKT细胞亚群相比比率显著增高。COA活化细胞主要是T细胞亚群。依据免疫荧光原理应用流式细胞术,为进一步研究NKT细胞结构和功能提供一种快速的检测手段。 相似文献
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流式细胞术在检测大肠埃希氏菌抗生素后效应中的应用 总被引:1,自引:0,他引:1
目的:探讨一种快速、准确检测细菌抗生素后效应(PAT)的流式细胞术.方法:选用碘化丙啶作为荧光染料,应用FACS Calibur型流式细胞仪(FCM)检测大肠埃希氏菌ATCC25922.以CellQuest软件采集、储存和分析数据.结果:清除加替沙星(GTLX 4MIC)270min后,流式细胞术可同时检测到菌体FSC,FL2增大.峰值右移,荧光强度增强.本方法是一种快速、多参数、理想的PAE检测手段. 相似文献
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Malignant glioma is one of the most common and deadly tumors in the central nervous system while developing effective treatments for this devastating disease remains a challenge. Previously, we demonstrated that the vitamin nicotinic acid (NA) inhibits glioma invasion. Here, we show that high-dose NA induces apoptosis of malignant glioma cells in vitro and in vivo. In cultured U251 glioma cells treated with NA, we detected ER stress that was likely caused by elevated intracellular calcium levels. The elevated calcium can be attributed to the activation of TRPV1, a cation channel that has been implicated in cutaneous flushing caused by NA administration. Our data further suggested that NA-induced apoptosis is mediated by the calcium-dependent proteases called calpains, whose activities are drastically upregulated by NA. NA-induced apoptosis of U251 cells can be attenuated by blocking calpain activity or knocking down TRPV1. These results reveal a novel function of NA in regulating glioma cell apoptosis via the calcium-dependent ER stress pathway and imply a potential application of NA for the treatment of malignant glioma. 相似文献
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XIRU XU YUAN FANG BIAO ZHANG SHICHAO TENG XIANG WU JING ZHANG XIAOQUN GU MEIXIA MA 《Biocell》2023,47(7):1595-1609
Background: While the Bushen Yizhi Formula can treat Alzheimer’s disease (AD), the yet to be ascertained
specific mechanism of action was explored in this work. Methods: Different concentrations of the Bushen Yizhi
Formula and amyloid-beta peptide (Aβ) were used to treat rat pheochromocytoma cells (P12) and human
neuroblastoma cells (SH-SY5Y). Cell morphological changes were observed to determine the in vitro cell damage. Cell
Counting Kit (CCK)-8 assay and flow cytometry were employed to identify cell viability and apoptosis/cell cycle,
respectively. Western blotting and immunohistochemistry were employed to measure the expressions of endoplasmic
reticulum stress (ERS)-related proteins (GRP78 and CHOP), p-IRE1α, IRE1α, ASK1, p-JNK, JNK, Bax, Bcl-2, XBP-1,
and Bim. Fura 2-acetoxymethyl ester (Fura-2/AM) was used to determine the intracellular calcium (Ca2+)
concentration. Also, an AD model was constructed by injecting Aβ into the CA1 area of the hippocampus in Sprague
Dawley rats. AD model rats were gavaged with different concentrations of Bushen Yizhi Formula for 14 consecutive
days. The Morris water maze experiment was conducted to test the learning and memory of rats. Hematoxylin &
Eosin (H&E) and Terminal-deoxynucleotidyl Transferase (TdT)-mediated dUTP Nick-End Labeling (TUNEL)
staining were done to determine histopathological changes in the brain. Results: Bushen Yizhi Formula relieved the
Aβ-induced effects including cell injury, decreased viability, increased apoptosis, G0/G1 phase cell cycle arrest, upregulation of GRP78, CHOP, p-IRE1α, p-JNK, Bax, XBP-1 and Bim, as well as down-regulation of Bcl-2. These
results were also seen with IRE1α silencing. While Aβ suppressed the learning and memory abilities of rats, the
Bushen Yizhi Formula alleviated these effects of Aβ. Brain nerve cell injury induced by Aβ could also be treated with
Bushen Yizhi Formula. Conclusion: Bushen Yizhi Formula could influence ERS through the IRE1α signaling pathway
to achieve its therapeutic effects on AD. 相似文献
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Serpil Ünver Saraydin Dursun Saraydin Zeynep Deniz ahin nan 《Microscopy research and technique》2020,83(4):436-445
Nowadays, apoptosis is mostly evaluated visually in histological studies. By using the quantitative digital image analysis, this study aimed to investigate the effect of acrylamide‐based monomers (acrylamide [AAm], methacrylamide [MAAm], N‐isopropylacrylamide [NIPAm]) on the cerebrum tissues in rats, which are the most common water‐soluble monomers in the production of polymeric hydrogels used as biomaterials. The Wistar albino rats weighing ~220–240 g were divided into control and three test groups. The control group received 1 mL of saline, and the test groups received 1 mL of aqueous 50 mg/kg/day intramuscular injection of AAm, MAAm, and NIPAm, respectively. At the end of the experiments, brain tissues of all rats euthanized by intramuscular injection of sodium pentobarbital were removed. Terminal deoxynucleotide transferase dUTP nick and labeling (TUNEL) method was applied to brain tissue sections. The monomers have been shown to cause apoptosis due to oxidative stress in cerebrum tissue. Based on apoptosis by tunneling method, quantitative digital image analysis of cell fragments was performed with Olympus cellSens Dimension 1.15 software, and the number, total count area, selected area, average area, and ROI% values of the fragments were found. In addition, the total area and ROI% values of the fragments increased linearly with increasing the molar mass of monomers from the digital image analysis data. Quantitative digital image analysis can facilitate the monitoring of apoptosis caused by the oxidative stress of monomers used in the production of the biomaterials. 相似文献
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Work materials experience a broad range of strains, strain rates, and temperatures in many manufacturing processes such as machining, forming, etc. Strain rate has an important effect on the yield and flow stress of work materials, especially metals, since at higher strain rates there is less time for thermally activated events; consequently, it is equivalent to a lowering of the temperature of the materials. On the other hand, it is also true that, for high strain rate deformations such as metal cutting, adiabatic plastic flow may produce significant temperature changes in the materials. Flow stress is significantly affected by the strain rate history; hence, mechanical behavior may not be fully described in terms of a mechanical equation of state relating the instantaneous stress, strain, strain rate, and temperature.Based on the concept of dislocation mechanics, a micromechanical approach with the new concept of temperature coefficient has been explored to overcome the model issues such as negative or constant flow stress above the critical temperatures. The flow stresses of aluminum 6061-T6 and titanium Ti-6Al-4V have been predicted, for the first time, using the modified micromechanical model based on the available baseline high strain rates test data. The constitutive model was further modified and extended to predict flow stress below as well as above the critical temperature. The corresponding model predictions were compared with the experimental data, attaining good agreement. 相似文献
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The pathogenesis of high altitude-related gastric mucosal injury remains poorly understood, this study aimed to
investigate the role of autophagy in hypoxia-induced apoptosis of rat gastric mucosal cells. Rats were randomized into
four groups which were maintained at an altitude of 400 m (P) or received no treatment (H), autophagy inducer
rapamycin (H+AI) or autophagy inhibitor 3-MA (H+AB) at an altitude of 4,300 m for 1, 7, 14 and 21 days,
respectively, and the morphology, ultrastructure, autophagy, and apoptosis of gastric mucosal tissues were examined.
Gastric mucosal epithelial cells CC-R039 were cultured under conditions of normoxia, 2% O2 (hypoxia), or 2%
O2+anti-mTORC1 for 0, 24, 48, and 72 h, respectively, and the autophagy and apoptosis were analyzed. CC-R039
cells were transfected with siHIF-1α, siTERT, or siRNA and the autophagy was examined. The results showed that the
exposure to hypoxia increased the autophagy and apoptosis of gastric mucosal cells in rats, and apoptosis was
aggravated by rapamycin treatment but alleviated by 3-MA treatment. Increased duration of hypoxia from 0 to 72 h
could increase the autophagy and apoptosis but decrease the proliferation of gastric mucosal cells. Inhibition of
mTORC1 with rapamycin led to further increase in apoptosis and even substantial cell death, and inhibition of HIF-
1α and TERT increased mTORC1 expression and reduced autophagy. Moreover, the inhibition of HIF-1α reduced
TERT expression. In conclusion, hypoxia could induce apoptosis of rat gastric mucosal cells by activating autophagy
through HIF-1α/TERT/mTORC1 pathway 相似文献