1.Effect of electromagnetic pulse exposure on permeability of blood-testicle barrier in mice.
Xiao-Wu WANG ; Gui-Rong DING ; Chang-Hong SHI ; Tao ZHAO ; Jie ZHANG ; Li-Hua ZENG ; Guo-Zhen GUO
Biomedical and Environmental Sciences 2008;21(3):218-221
OBJECTIVETo study the effect of electromagnetic pulse (EMP) exposure on the permeability of blood-testicle barrier (BTB) in mice.
METHODSAdult male BALB/c mice were exposed to EMP at 200 kV/m for 200 pulses with 2 seconds interval. The mice were injected with 2% Evans Blue solution through caudal vein at different time points after exposure, and the permeability of BTB was monitored using a fluorescence microscope. The testis sample for the transmission electron microscopy was prepared at 2 h after EMP exposure. The permeability of BTB in mice was observed by using Evans Blue tracer and lanthanum nitrate tracer.
RESULTSAfter exposure, cloudy Evans Blue was found in the testicle convoluted seminiferous tubule of mice. Lanthanum nitrate was observed not only between testicle spermatogonia near seminiferous tubule wall and sertoli cells, but also between sertoli cells and primary spermatocyte or secondary spermatocyte. In contrast, lanthanum nitrate in control group was only found in the testicle sertoli cells between seminiferous tubule and near seminiferous tubule wall.
CONCLUSIONEMP exposure could increase the permeability of BTB in the mice.
Animals ; Blood-Testis Barrier ; metabolism ; radiation effects ; Coloring Agents ; Electromagnetic Fields ; Evans Blue ; Lanthanum ; Male ; Mice ; Mice, Inbred BALB C ; Permeability ; radiation effects ; Seminiferous Tubules ; metabolism ; radiation effects
2.Iron mediates endothelial cell damage and blood-brain barrier opening in the hippocampus after transient forebrain ischemia in rats.
Sun Mi WON ; Jin Hwan LEE ; Ui Jin PARK ; Jina GWAG ; Byoung Joo GWAG ; Yong Beom LEE
Experimental & Molecular Medicine 2011;43(2):121-128
Blood cells are transported into the brain and are thought to participate in neurodegenerative processes following hypoxic ischemic injury. We examined the possibility that transient forebrain ischemia (TFI) causes the blood-brain barrier (BBB) to become permeable to blood cells, possibly via dysfunction and degeneration of endothelial cells in rats. Extravasation of Evans blue and immunoglobulin G (IgG) was observed in the hippocampal CA1-2 areas within 8 h after TFI, and peaked at 48 h. This extravasation was accompanied by loss of tight junction proteins, occludin, and zonula occludens-1, and degeneration of endothelial cells in the CA1-2 areas. Iron overload and mitochondrial free radical production were evident in the microvessel endothelium of the hippocampus before endothelial cell damage occurred. Administration of deferoxamine (DFO), an iron chelator, or Neu2000, an antioxidant, blocked free radical production and endothelial cell degeneration. Our findings suggest that iron overload and iron-mediated free radical production cause loss of tight junction proteins and degeneration of endothelial cells, opening of the BBB after TFI.
Animals
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Blood-Brain Barrier/*metabolism
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Capillary Permeability
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Endothelial Cells/*metabolism
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Evans Blue/metabolism
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Free Radicals/metabolism
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Hippocampus/*metabolism/pathology
;
Iron/*metabolism
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Ischemic Attack, Transient/pathology/*physiopathology
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Male
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Membrane Proteins/metabolism
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Rats
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Rats, Sprague-Dawley
3.Tanshinone ⅡA activates PI3K/AKT signaling pathway to inhibit the apoptosis of mice cochlear pericytes induced by high glucose.
Tian Feng SHI ; Jin Jing JIA ; Tian Lan HUANG ; Jing Wen MA ; Jun Qiang SI ; Ke Tao MA ; Li LI
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2023;58(7):681-689
Objective: To investigate whether tanshinone ⅡA can protect the apoptosis of mice cochlear pericytes induced by high glucose and its specific protective mechanism, so as to provide experimental evidence for the prevention and treatment of diabetic hearing loss. Methods: C57BL/6J male mice were used to prepare type 2 diabetes model, which were divided into normal (NG) group, diabetic (DM) group, diabetic+tanshinone ⅡA (HG+tanshinone ⅡA) group and tanshinone ⅡA group. Each group had 10 animals. Primary cochlear pericytes were divided into NG group, HG group (high glucose 35 mmol/L), HG+tanshinone ⅡA (1, 3, 5 μmol/L) group, HG+Tanshinone ⅡA+LY294002 (PI3K/AKT pathway inhibitor) group, LY294002 group, tanshinone ⅡA group and DMSO group. Auditory brainstem response (ABR) was used to measure hearing threshold. Evans blue was used to detect the permeability of blood labyrinth barrier in each group. TBA methods were used to detect oxidative stress levels in various organs of mice. Morphological changes of stria vascularis were observed by hematoxylin-eosin staining (HE). Evans blue was used to detect the vascular labyrinth barrier permeability in cochlea. The expression of apoptosis protein in stria vascularis pericytes was observed by immunofluorescence. Pericytes apoptosis rate was observed by flow cytometry. DCFH-DA was combined with flow cytometry to detect intracellular ROS content, and Western blot was used to detect the expression of apoptotic proteins (Cleaved-caspase3, Bax), anti-apoptotic proteins (BCL-2) and pathway proteins (PI3K, p-PI3K, AKT, p-AKT). SPSS software was used for statistical analysis. Independent sample t test was performed, and P<0.05 was considered statistically significant. Results: Animal experiments: Tanshinone ⅡA decreased the hearing threshold of DM group [(35.0±3.5) dB SPL vs. (55.3±8.1) dB SPL] (t=4.899, P<0.01), decreased the oxidative stress level in cochlea (t=4.384, P<0.05), improved the structure disorder, atrophy of cochlea vascular lines, vacuole increased phenomenon. Tanshinone ⅡA alleviated the increased permeability of the blood labyrinth barrier [Evans blue leakage (6.84±0.27) AU vs. (8.59±0.85) AU] in the cochlea of DM mice (t=2.770, P<0.05), reversed the apoptotic protein: Caspase3 (t=4.956, P<0.01) and Bax (t=4.388, P<0.05) in cochlear vascularis. Cell experiments: Tanshinone ⅡA decreased intracellular ROS content in a concentration-dependent way (t=3.569, P<0.05; t=4.772, P<0.01; t=7.494, P<0.01); Tanshinone ⅡA decreased apoptosis rate and apoptotic protein, and increased the expression of anti-apoptotic protein, p-PI3K/PI3K and p-AKT/AKT in concentration-dependent manner (all P values<0.05); LY294002 reversed the protective effect of tanshinone ⅡA on pericytes apoptosis (all P values<0.05). Conclusion: Tanshinone ⅡA can inhibit the apoptosis of cochlear pericytes induced by high glucose by reducing oxidative stress level and activating PI3K/AKT signaling pathway under high glucose environment, thus playing a protective role in diabetic hearing loss.
Animals
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Male
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Mice
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Apoptosis
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bcl-2-Associated X Protein
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Diabetes Mellitus, Type 2
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Evans Blue
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Glucose
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Hearing Loss
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Mice, Inbred C57BL
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Pericytes/metabolism*
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Phosphatidylinositol 3-Kinases/metabolism*
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Proto-Oncogene Proteins c-akt/metabolism*
;
Reactive Oxygen Species/metabolism*
;
Signal Transduction
4.Research on the relationship between thick greasy tongue fur formation and vascular endothelial cell permeability with the protein expression of zonula occludens-1.
Wen-Jie QI ; Miao-Miao ZHANG ; Hong WANG ; Yan WEN ; Bao-En WANG ; Shu-Wen ZHANG
Chinese journal of integrative medicine 2011;17(7):510-516
OBJECTIVETo determine the relationship of thick greasy tongue fur formation and permeability of vascular endothelial cells (ECs) with the protein expression of zonula occludens-1 (ZO-1).
METHODSSprague Dawley rats were randomly divided into a model group of severe acute pancreatitis (SAP) and a sham-operated (SO) group. The SAP rats were further divided into two subgroups on the basis of tongue-coating status: a thick greasy tongue fur group (SAP-TGF) and a normal tongue fur group (SAP-NF). Six rats were chosen randomly from every group mentioned above for an Evans blue assay 5 days after model establishment. For the histomorphology analysis, the expressions of ZO-1 protein and mRNA were studied by hematoxylin-eosin (H&E) staining, transmission electron microscope, Western blot, and Q-PCR using blood and tongue tissues, which were collected from 8 rats randomly chosen from each group.
RESULTSThe papillae density of the rat tongue surface and the caryocinesis frequency of the basal layer were significantly increased in the SAP-TGF group compared with the SO group (P<0.05). Evans blue levels in the tongue tissue of the SAP-TGF group were significantly higher than that of the SO and SAP-NF groups (P<0.05). Vascular ECs were wider and obviously fissured in the SAP-TGF group under transmission electron microscope observation. The protein and mRNA expression of ZO-1 in the SAP-TGF group were lower than those in the SAP-NF (P<0.05).
CONCLUSIONSReproductive activity enhancement of glossal epithelial cells was one of the main characteristics of thick greasy tongue fur formation. An increase in vasopermeability was closely associated with thick greasy tongue fur formation. Tight junction structural variation of vascular ECs might play an important role in the pathological and physiological process of thick greasy tongue fur formation.
Animals ; Blotting, Western ; Cell Membrane Permeability ; Endothelial Cells ; cytology ; Evans Blue ; metabolism ; Gene Expression Regulation ; Male ; Membrane Proteins ; genetics ; metabolism ; Phosphoproteins ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tongue ; pathology ; ultrastructure ; Zonula Occludens-1 Protein
5.Effect of hyaluronic acid chitosan-based microemulsion on the permeability of blood brain barrier in mice.
Jing YAO ; Jian-ping ZHOU ; Qi-neng PING ; Yun LU ; Li YU
Acta Pharmaceutica Sinica 2006;41(7):615-618
AIMTo study the effect of hyaluronic acid chitosan-based microemulsion (HAC-ME) on the permeability of blood brain barrier( BBB) by using Evans blue (EB) as the indicator.
METHODSA formamide extraction-ultraviolet spectrophotometry method was employed to determine the concentrations of EB in each of the tissues. The in vivo distribution of HAC-ME groups containing EB in mice and the fluorescence intensity and diffusion domain of brain slices were all studied.
RESULTSContrasting to the common microemulsion (ME), HAC-ME at the lower concentration of HAC (<5 mg x mL(-1)) could further improve the transporting of EB across the BBB while EB concentration in other tissues decreased, and Tmax was delayed about 30 min.
CONCLUSIONHAC-ME could facilitate the transporting of EB across the BBB and it was concentration dependent. While the brain targeting absorptive capability of HAC-ME was enhanced.
Animals ; Blood-Brain Barrier ; drug effects ; metabolism ; Brain ; metabolism ; Chitosan ; chemistry ; pharmacology ; Dose-Response Relationship, Drug ; Emulsions ; Evans Blue ; chemistry ; pharmacokinetics ; Female ; Hyaluronic Acid ; chemistry ; pharmacokinetics ; pharmacology ; Male ; Mice ; Particle Size ; Tissue Distribution
6.Ultrasound-mediated microbubble destruction increases capillary permeability in rat skeletal muscles.
Yi LAO ; Jian-cheng XIU ; Chang-lian XIE ; Xiang-hui CHEN ; Jue-fei WU ; Jian-ping BIN ; Yi-li LIU
Journal of Southern Medical University 2008;28(4):542-544
OBJECTIVETo investigate the effects of ultrasound mediated microbubble destruction on capillary permeability in rat skeletal muscles.
METHODSEighteen SD rats were randomized into 3 groups, namely the Evans blue (EB) group, EB+ultrasound (E+U) group and EB+microbubble+ultrasound (U+E+M) group with corresponding treatments, using EB injected into the carotid artery as the indicator for capillary permeability. The microbubbles were injected through the carotid artery with fixed ultrasound parameters. The spillover of EB was estimated under fluorescence microscope according to the visual staining scores. The contents of EB in the skeletal muscles were calculated according to the standard curve and spectrophotometry.
RESULTSEB spillover was observed around the capillaries in E+U+M group, which had a significantly higher visual score than EB group and E+U group (0 and 0-1, respectively, P<0.05). The EB content was 51.57-/+3.89 microg/g in E+U+M group, also significantly higher than those in EB group (28.99-/+4.67 microg/g) and E+U group (30.99-/+4.11 microg/g) (P<0.05).
CONCLUSIONExposure to both ultrasound and microbubble contrast agents results in increased capillary permeability of rat skeletal muscles, which might be an important mechanisms for gene delivery enhancement by ultrasound contrast agents.
Animals ; Capillary Permeability ; physiology ; Coloring Agents ; administration & dosage ; pharmacokinetics ; Contrast Media ; administration & dosage ; Evans Blue ; administration & dosage ; pharmacokinetics ; Female ; Male ; Microbubbles ; Microscopy, Fluorescence ; Muscle, Skeletal ; blood supply ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spectrophotometry ; Ultrasonics
7.Effect of RMP-7 and its derivatives on the transportation of liposome into the brain.
Xiao-bin ZHANG ; Yi-guang JIN ; Ying XIE ; Kun XU ; Xin-pu HOU
Acta Pharmaceutica Sinica 2003;38(11):867-870
AIMTo study the action of RMP-7 and its derivative on transporting liposome across the blood brain barrier (BBB) into the brain.
METHODSRMP-7 and DSPE-PEG-NHS [[1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-n-[poly (ethylene-glycol)]-hydroxy succinamide]] were conjugated together in mild condition and MALDI-TOF-MS (Matrix-Assisted Laser Desorption-Ionization Time-of-Flight Mass Spectrometry) was used to determine their molecular ratio. An in vitro BBB model was established and used to determine in vitro bioactivity of RMP-7 and its derivative. The fluorescence of brain slices and the Evens Blue (EB) concentration in the brain, liver, spleen, lung and kidney of each group were used to evaluate the in vivo bioactivity of RMP-7 and its derivative on transporting liposome across the BBB.
RESULTSThe average molecular weight (MW) of the reaction product was 4,900, while those of DSPE-PEG-NHS and RMP-7 were 3,224 and 1,098. The results demonstrated that RMP-7 was conjugated to DSPE-PEG-NHS at the molecular ratio of 1:1, so the product was DSPE-PEG-RMP-7. RMP-7 and DSPE-PEG-RMP-7 was shown to improve the transporting of peralcohol enzyme across the in vitro BBB model 2-3 times higher than the peralcohol enzyme only. DSPE-PEG-RMP-7 could facilitate the transporting of EB into brain more easily than RMP-7.
CONCLUSIONBoth RMP-7 and DSPE-PEG-RMP-7 could facilitate the transporting of liposome across the BBB, especially DSPE-PEG-RMP-7.
Animals ; Biological Transport ; Blood-Brain Barrier ; drug effects ; Bradykinin ; analogs & derivatives ; pharmacology ; Brain ; metabolism ; Drug Carriers ; Drug Delivery Systems ; Evans Blue ; pharmacokinetics ; Liposomes ; pharmacokinetics ; Phosphatidylethanolamines ; Polyethylene Glycols ; Rats ; Rats, Sprague-Dawley ; Tissue Distribution
8.Protective effect of jiedu tongluo injection on cerebral edema in rats with lesion of cerebral ischemia/reperfusion.
Li-Fei WU ; Yue XING ; Ya-Lan GUAN ; Zhen-Quan LIU ; Wen-Sheng ZHANG
China Journal of Chinese Materia Medica 2014;39(6):1088-1092
OBJECTIVETo investigate the protective effects of Jiedu Tongluo injection on cerebral edema induced by focal lesion of cerebral ischemia/reperfusion, the hydrous content of brain and the expressions of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), E-selectin and MMP-9 in rats.
METHODThe model of brain middle cerebral artery ischemia/reperfusion was established by the thread approach. After 24 hours of reperfusion, cerebral edema formation was determined by the hydrous content of brain. The permeability of blood brain barrier was evaluated based on the leakage of Evans blue. Enzyme-linked immunoadsordent assay (ELISA)was used to examine the expression of ICAM-1, VCAM-1, E-selectin. The expression of MMP-9 was measured by immunohistochemistry.
RESULTJDTL, in the dose of 2 mL x kg(-1) and 4 mL x kg(-1), relieved cerebral edema (P < 0.05, P < 0.01), reduced the expressions of ICAM-1, VCAM-land E-selectin and decreased MMP-9 activity (P < 0. 05, P < 0.01) in model rats.
CONCLUSIONJiedu Tongluo injection has a protective effect on rat brain from cerebral edema induced by the injury of focal cerebral ischemia/reperfusion. The mechanism is related to that Jiedu Tongluo injection can reduce the expressions of ICAM-1, VCAM-1 and E-selectin and inhibit of MMP-9 activation in rat brain.
Animals ; Blood-Brain Barrier ; drug effects ; metabolism ; Brain Edema ; etiology ; metabolism ; prevention & control ; Brain Ischemia ; complications ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; E-Selectin ; metabolism ; Evans Blue ; metabolism ; Gene Expression Regulation, Enzymologic ; drug effects ; Injections ; Intercellular Adhesion Molecule-1 ; metabolism ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Permeability ; drug effects ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; complications ; Vascular Cell Adhesion Molecule-1 ; metabolism
9.Schisandrin B Inhibits NLRP3 Inflammasome Pathway and Attenuates Early Brain Injury in Rats of Subarachnoid Hemorrhage.
Song CHEN ; Yi-Hang DING ; Song-Sheng SHI ; Xian-Kun TU
Chinese journal of integrative medicine 2022;28(7):594-602
OBJECTIVE:
To determine whether Schisandrin B (Sch B) attenuates early brain injury (EBI) in rats with subarachnoid hemorrhage (SAH).
METHODS:
Sprague-Dawley rats were divided into sham (sham operation), SAH, SAH+vehicle, and SAH+Sch B groups using a random number table. Rats underwent SAH by endovascular perforation and received Sch B (100 mg/kg) or normal saline after 2 and 12 h of SAH. SAH grading, neurological scores, brain water content, Evan's blue extravasation, and terminal transferase-mediated dUTP nick end-labeling (TUNEL) staining were carried out 24 h after SAH. Immunofluorescent staining was performed to detect the expressions of ionized calcium binding adapter molecule 1 (Iba-1) and myeloperoxidase (MPO) in the rat brain, while the expressions of B-cell lymphoma 2 (Bcl-2), Bax, Caspase-3, nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3), apoptosis-associated specklike protein containing the caspase-1 activator domain (ASC), Caspase-1, interleukin (IL)-1β, and IL-18 in the rat brains were detected by Western blot.
RESULTS:
Compared with the SAH group, Sch B significantly improved the neurological function, reduced brain water content, Evan's blue content, and apoptotic cells number in the brain of rats (P<0.05 or P<0.01). Moreover, Sch B decreased SAH-induced expressions of Iba-1 and MPO (P<0.01). SAH caused the elevated expressions of Bax, Caspase-3, NLRP3, ASC, Caspase-1, IL-1β, and IL-18 in the rat brain (P<0.01), all of which were inhibited by Sch B (P<0.01). In addition, Sch B increased the Bcl-2 expression (P<0.01).
CONCLUSION
Sch B attenuated SAH-induced EBI, which might be associated with the inhibition of neuroinflammation, neuronal apoptosis, and the NLRP3 inflammatory signaling pathway.
Animals
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Apoptosis
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Brain/pathology*
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Brain Injuries/pathology*
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Caspase 3/metabolism*
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Cyclooctanes
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Evans Blue
;
Inflammasomes/metabolism*
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Interleukin-18/metabolism*
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Lignans
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NLR Family, Pyrin Domain-Containing 3 Protein/metabolism*
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Polycyclic Compounds
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Proto-Oncogene Proteins c-bcl-2/metabolism*
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Rats
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Rats, Sprague-Dawley
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Subarachnoid Hemorrhage/drug therapy*
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Water
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bcl-2-Associated X Protein/metabolism*