Effects of nano lead oxide exposure on cognitive function and inflammatory mechanism of leukocyte infiltration in mice / 中华行为医学与脑科学杂志

Objective:To investigate the effect of nano lead oxide (nano-PbO) exposure on learning and memory as well as spatial exploration ability in the mice, and the role of leukocyte infiltration of brain tissue in neurobehavioral damage caused by nano-PbO exposure.Methods:A total of 60 male SPF grade Kunming mice were divided into control group, low-dose nano-PbO group, medium-dose nano-PbO group and high-dose nano-PbO group according to body mass matching method, with 15 mice in each group.Mice in low, medium and high dose groups of nano-PbO were intraperitoneally injected with 5 mg·kg -1, 10 mg·kg -1, 20 mg·kg -1 nano-PbO, respectively. And mice in the control group were intraperitoneally injected with the same volume of 0.9% normal saline.The frequency of intervention was once a day for 28 days.Morris water maze test and open field test were used to detect the ability of learning and memory and spatial exploration of mice. Western blot was used to detect the protein expression of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) in hippocampus of mice, intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) in mouse microvessels and lymphocyte function-associated antigen-1 (LAF-1) in mouse blood leukocyte. The proportion of leukocytes in mouse brain was detected by flow cytometry. All statistical analyses were performed by SPSS 20.0. Morris water maze data were analyzed by repeated measures ANOVA, the other data among multiple groups were compared by one-way ANOVA, and Tukey's test was used for further pairwise comparison.Pearson correlation analysis was performed to evaluate the correlation between neurobehavioral indexes and the proportion of white blood cells, TNF-α and IL-1β in brain tissue. Results:Morris water maze results showed that the escape latency of the four groups of mice had a significant interaction between group and time( F=3.21, P<0.05). The escape latencies of mice in middle and high dose groups of nano-PbO were higher than that in the control group (both P<0.05), and the numbers of crossing the platform of the two groups were lower than that in the control group (both P<0.05). The results of open field test showed that there was a statistically significant difference in the residence time of the mice in the four groups ( F=119.10, P<0.01). The total standing times of mice in the middle group and high dose group of nano-PbO were lower than that in the control group (both P<0.01). The results of Western blot showed that the levels of TNF-α and IL-1β in hippocampus tissue of mice were significant differences among the four groups ( F=7.21, 9.89, both P<0.05). The levels of TNF-α and IL-1β in the hippocampus of mice in the high-dose nano-PbO group were higher than those in the control group (TNF-α: (0.35±0.10), (1.03±0.30), P<0.05; IL-1β: (0.32±0.10), (0.50±0.15), P<0.05). The results of flow cytometry analysis showed that the proportions of leukocytes in the brain tissue of mice in the low, medium and high dose groups of nano-PbO were (9.99±1.09)%, (13.03±0.94)% and (16.51±3.89)%, respectively. Among them, the proportions of leukocytes in the middle and high dose groups of nano-PbO were significantly higher than that in the control group((8.13±1.29)%) (both P<0.05). The results of correlation analysis showed that the proportion of leukocytes, levels of TNF-α, IL-1β protein of hippocampus in the medium, high dose groups of nano-PbO were negatively correlated with the behavioral indexes ( r=-0.815, -0.744, -0.578, all P<0.01; r=-0.771, -0.836, -0.704, all P<0.05; r=-0.823, -0.876, -0.695, all P<0.05). The results of Western blot showed that the levels of ICAM-1 and VCAM-1 in cerebral microvessels of mice in the four groups were significantly different ( F=5.51, 16.19, both P<0.05). The levels of ICAM-1 and VCAM-1 in the middle and high dose groups of nano-PbO were higher than those in the control group(ICAM-1: (1.07±0.16), (1.21±0.35), (0.59±0.19), all P<0.05; VCAM-1: (0.68±0.12), (1.92±0.23), (0.23±0.05), both P<0.05). In addition, there was a significant difference in the level of LFA-1 protein in blood leukocytes of mice in the four groups ( F=41.80, P<0.05). The levels of LFA-1 in the middle and high dose groups of nano-PbO were higher than that in the control group((0.33±0.06), (0.89±0.23), (0.05±0.01), both P<0.05). Conclusion:The nano-PbO exposure can lead to cognitive impairment and increased inflammatory factors in the hippocampus of mice, which may be related to the increase of ICAM-1 and VCAM-1 in vascular endothelial cells, which promotes leukocyte infiltration into brain tissue.

Therapeutic effects of the extract of Sancao Formula, a Chinese herbal compound, on imiquimod-induced psoriasis via cysteine-rich protein 61 / 中西医结合学报

OBJECTIVE@#Psoriasis is a common chronic inflammatory skin disease that is prone to recurrence, and the proinflammatory factor, cysteine-rich protein 61 (Cyr61), is important in its pathophysiology. Long-term clinical practice has shown that Sancao Formula (SC), a Chinese herbal compound, is effective in the treatment of psoriasis, but the precise mechanism remains unknown. In this study, we investigate the mechanism by which SC extract alleviates imiquimod (IMQ)-induced psoriasis.@*METHODS@#The expression of Cyr61 in psoriatic lesions and normal healthy skin was detected using immunohistochemical analysis to investigate the biological role of Cyr61 in models of psoriatic inflammation. A psoriatic mouse model was established by topical application of IMQ, and the effect of topical application of SC extract was evaluated using the psoriasis area and severity index (PASI) score, hematoxylin-eosin staining, and histopathological features of the skin. Next, a HaCaT cell inflammation model was established using interferon-γ (IFN-γ), and the effect of SC extract on the mRNA and protein levels of Cyr61 and intercellular cell adhesion molecule-1 (ICAM-1) was confirmed using Western blot and quantitative real-time polymerase chain reaction analyses.@*RESULTS@#Immunohistochemical staining showed that the expression of Cyr61 in psoriatic lesions was higher than that in normal skin samples (78.26% vs 41.18%, P < 0.05), and the number of Cyr61-positive cells in psoriatic lesions was also significantly higher than in normal skin (18.66 ± 2.51 vs 4.33 ± 1.52, P < 0.05). Treatment in mice with IMQ-induced psoriasis showed that SC extract could significantly improve the inflammatory phenotype, PASI score (10.875 ± 0.744 vs 3.875 ± 0.582, P < 0.05), and pathological features compared with those in IMQ model group; SC treatment was also associated with decreased levels of Cyr61 and ICAM-1. In the IFN-γ-induced inflammatory cell model, the mRNA and protein levels of Cyr61 and ICAM-1 were upregulated, while the SC extract downregulated the levels of Cyr61 and ICAM-1.@*CONCLUSION@#The results provide a theoretical basis for the involvement of Cyr61 in the pathogenesis of psoriasis, and suggest that SC should be used to target Cyr61 for the prevention of psoriasis recurrence.

Cornuside ameliorated experimental autoimmune encephalomyelitis by limiting the recruitment of CD4+ T lymphocytes in the spinal cord

We conducted this study to determine whether cornuside could improve the neurological deficit symptoms of experimental autoimmune encephalomyelitis (EAE) rats, as well as determine the potential involvement of CD4+ T lymphocytes, vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), and tumor necrosis factor-α (TNF-α). Altogether, 32 Lewis rats were randomly divided into control, EAE, EAE/prednisolone, and EAE/cornuside, wherein their neurological function was assessed every day. CD4+ T lymphocyte recruitment into the spinal cord (SC) was evaluated using immunohistochemistry. The VCAM-1, ICAM-1 and TNF-α mRNA expressions in the SC were determined by real-time quantitative PCR, and the VCAM-1 and ICAM-1 proteins were determined by western blotting. Compared to the control group, the EAE group rats with neurological deficits had enhanced CD4+ T lymphocyte infiltration and higher expression levels of VCAM-1, ICAM-1, and TNF-α in the SC. Meanwhile, compared with the EAE group, the EAE/cornuside and EAE/prednisolone groups had lower neurological scores, less CD4+ T lymphocyte infiltrations, and lower expression levels of VCAM-1, ICAM-1, and TNF-α in the SC. Thus, cornuside ameliorated EAE, which could be owed to the inhibition of CD4+ T lymphocyte recruitment and VCAM-1, ICAM-1, and TNF-α expressions in the SC

Effect of Modified Danggui Shaoyao San on MMP-2, ICAM-1, Emorheology and Inflammation in Patients with Chronic Pelvic Inflammatory Disease / 中国实验方剂学杂志

Objective: To explore the effect of modified Danggui Shaoyao San on matrix metalloproteinase-2(MMP-2), intercellular adhesion molecule-1 (ICAM-1), emorheology and inflammation in patients with chronic pelvic inflammatory disease. Method: Patients with chronic pelvic inflammatory disease during May 2017 to June 2018 were randomly divided into treatment group and control group, with 37 cases in each group. The control group was orally treated with levofloxacin tablets and metronidazole tablets. In addition to the therapy of the control group, the treatment group was also given modified Danggui Shaoyao San. Traditional Chinese medicine(TCM) symptom scores, MMP-2 and ICAM-1, interleukin-1β(IL-1β), IL-6, IL-4, IL-10, and transforming growth factor-β (TGF-β), grain-megakaryocyte colony stimulating factor (GM-CSF), tumor necrosis factor-alpha (TNF-α), whole blood viscosity (ηb), plasma viscosity (ηp), erythrocyte aggregation index (AI), fibrinogen (Fib) before and after treatment were compared between two groups. The efficacy, adverse reactions and recurrence were observed in two groups. Result: The clinical efficacy of treatment group was better than that of the control group (Z=2.791, PPα, IL-1β, IL-6, GM-CSF,ηb,ηp, AI, Fib levels in the treatment group after treatment were significantly lower than those in control group (Pβ1 levels in treatment group after treatment were significantly higher than those in control group (Pχ2=6.198, PConclusion: Modified Danggui Shaoyao San has a significant clinical efficacy in the treatment of CPID, and can effectively relieve clinical symptoms and greatly reduce the recurrence rate, which may be related to the improvement of the regulation of MMP-2 and ICAM-1 levels, the inhibition of inflammatory reactions and the improvement of hemorheology.

TNF-α promotes the quantity and ICAM-1 expression of endothelial microparticles / 医用生物力学

Objective To study the role of cyclic strain-modulated tumor necrosis factor-α (TNF-α) played in the quantity and intercellular cell adhesion molecule-1(ICAM-1) expression of endothelial microparticles (EMPs). Methods The endothelial cells (ECs) primarily cultured from rat aorta were applied with 5% cyclic strain (to simulate normal physiological condition) and 18% cyclic strain (to simulate hyper-tension condition), respectively, by using FX-4000T cyclic stain loading system for 24 hours at the loading frequency of 1.25 Hz. The mRNA expression of TNF-α under different amplitudes of cyclic strain was determined by real time-PCR. The TNF-α was then used to stimulate the ECs from rat aorta, and the supernatants were collected and ultracentrifuged to get endothelial microparticles (EMPs), which were then identified by lipophilic styryl membrane staining and transmission electron microscope for morphological identification. The quantities of Annexin V positive EMPs under TNF-α stimulation were counted by flow cytometer and ICAM-1 expression on EMPs was detected as well. Results Compared with the 5% normal cyclic strain, under 18% high cyclic strain condition,the mRNA expression of TNF-α in ECs increased significantly. TNF-α could then significantly up-regulate the production of Annexin V positive EMPs and promote the expression of ICAM-1 on EMPs. Conclusions The over-expression of TNF-α in ECs under high cyclic strain might mediate the high production of EMPs and over-expression of ICAM-1 on EMPs. The research findings will provide new experiment evidence for further studying the role of EPCs in the mechanobiological mechanism of vascular remodeling.

Early intervention of Didang Decoction on macroangiopathy in type 2 diabetic rats and its mechanisms / 中草药

Objective: To observe the effects of the early intervention with Didang Decoction (DDD) on macroangiopathy in type 2 diabetic rats. Methods: The type 2 diabetic rat model was established using high-fat diet and Streptozotocin (STZ), and the rats were divided into control, model, Pioglitazone (2.7 mg/kg), Simvastatin (1.8 mg/kg), early-, mid-, and late-term DDD-intervene groups (ig administered with 3.24 g/kg DDD once daily, before 4 weeks, at the same time, and after 4 weeks of STZ administration, respectively), the rats in each group were administered until 24 weeks of STZ administration. The immunohistochemical and pathological changes of intercellular cell adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in the aorta were observed, and the expression of nuclear factor-kappa B (NF-κB) and matrix metalloproteinases-9 (MMP-9) in aorta was detected using Western blotting. Results: Compared with the model group, the ICAM-1 expression decreased in early- and mid-term DDD-intervene groups, and Simvastatin group (P < 0.05), the VCAM-1 expression decreased in early-term DDD-intervene and Simvastatin groups (P < 0.05), the protein expression of NF-κB and MMP-9 was lower in early- and mid-term DDD-intervene groups (P < 0.05), and it is the most obvious in early-term DDD-intervene group. Conclusion: The contents of ICAM-1 and VCAM-1 as well as the protein expression of NF-κB and MMP-9 in type 2 diabetic rats decrease after early-term DDD-intervene, which could regulate NF-κB signaling pathway to delay the development of diabetic macroangiopathy.

Effects of polydatin on intercellular cell adhesion molecule-1 in cortex of neonatal rats with hypoxicischemia brain damage / 国际儿科学杂志

Objective To investigate the protective effects and possible mechanisms of polydatin(PD) on hypoxic-ischemia brain damage(HIBD) in neonatal rat by means of the expression of intercellular cell adhesion molecule( ICAM)-1 in cortex. Methods Fifty-four SD rats were divided into 3 groups at random, shame group (no HIBD), HIBD group (no medication) ,and PD treatment group. 7day-old rat's HIBD model was established by Rice's method. ICAM-1 expression in brain after HIBD was measured in different time by Immunohistochemitry technique. Results In sham group, there were less brain microvessel immunostained positively. In HIBD group,the number of ICAM-1 immuno-positive staining blood vessels increased significantly after 6h, 12h reached peak point. ICAM-1 immunoreactive staining of blood vessels levels continued in the peak after 24h. In PD treatment group, ICAM-1 expression on brain microvascular endothelial decreased after HIBD 6h, 12h, 24h, which was significant compared with HIBD group( P < 0. 05 or P < 0. 01 separately). Conclusion The expression of ICAM-1 was involved in the procedure induced by hypoxic-ischemia. After HIBD, polydatin would downregulate ICAM-1 expression in cerebral microvascular endothelial, and inhibite the inflammatory response.

Immunohistochemical Study of Immune Cells, with a Special Emphasis on Macrophage Subpopulations in the Rat Thymus after Cyclophosphamide Treatment / 대한해부학회지

This study was undertaken to investigate the in vivo effects of cyclophosphamide (CY) on immune cells, with a special emphasis on macrophage subpopulations in the thymus of rats. After a single dose of CY (150 mg/kg) was administered to Sprague-Dawley rats by intraperitoneal injection, the rats were sacrificed at days 1, 3, 7, 14 and 28. The immunohistochemical characterization of the tissues were carried out using various monoclonal antibodies in cryostat-cut sections. CD4(+/-) and CD8(+/-) T cells were greatly decreased in number after CY treatment. However, macrophages, including the ED1(+/-) ED2(+/-) and ED3(+/-) macrophages exhibited signs of cellular activation such as an increase in number and size of cell, and an upregulation of the ED1, ED2 and ED3 reactive surface molecule expression. Contrarily, CY elicited a decrease in number of thymic dendritic cells (DCs). CY induced a conspicuous upregulation of ICAM-1 expression in the thymic cortex. Most of these features began to detectable from the first day and reached the maximun on the third and seventh days, but two weeks after CY administration, these phenomena began to disap. In conclusion, the results of the present study shed more light on the effects of CY on various subpopulations of macrophages and other types of immune cells and on ICAM-1 expression in the rat thymus.

Immunohistochemical Study of Immune Cells, with a Special Emphasis on Macrophage Subpopulations in the Rat Spleen after Cyclophosphamide Treatment / 대한해부학회지

This study was undertaken to investigate the in vivo effects of cyclophosphamide (CY) on subpopulations of macrophages and other types of immune cells including dendritic cells (DCs) as well as on ICAM-1 expression in the spleen of rats. After a single dose of CY (150 mg/kg) was administered to Sprague-Dawley rats by intraperitoneal injection, the rats were sacrificed at 1, 3, 7, 14 and 28 days. The immunocytochemical characterization of the tissues were carried out using the monoclonal antibodies W3/25, OX8, HIS24, 8A2, OX6, OX62, ED1, ED2, ED3, and TLD-4C9 for analysis of macrophage subpopulations, DC(s), CD4(+) and CD8(+) T cells, B cells and ICAM-1 expression in cryostat-cut sections. CY exhibited a profound immunosuppressive effect on CD4(+) and CD8(+) T cells as well as B cells as was expected. However, it was found that CY induced an increase in number of certain subpopulations of macrophages, including ED1(+), ED2(+) and ED3(+) macrophages. Contrarily, CY elicited a decrease in number of DCs. CY induced a conspi-cuous upregulation of ICAM-1 on certain populations of leukocytes. This increased expression of ICAM-1 after CY treatment appears to be related with the recruitment of certain populations of leukocytes. Most of these features began to appear from the first day and reached the maximun on the third and especially, the seventh day, but two weeks after CY administration, these phenomena declined. In conclusion, the present study provided a new insight into the differential effects of CY on various populations and subpopulations of immune cells in the rat spleen.