1.Correlation between multi -voxel magnetic resonance spectroscopy parameter choline/creatine and distribution of glioma stem cells
Xyulei HU ; Tao HE ; Xilong WANG ; Gaoyang QI ; Xinghao DUAN ; Hui MA
Chinese Journal of Neuromedicine 2017;16(6):585-590
Objective To investigate the correlation between mutiple-voxel magnetic resonance spectroscopy (1H-MRS) parameter choline/creatine (Cho/Cr) and distribution of glioma stem cells (GSCs). Methods Sixteen patients with high-grade glioma approved by pathology, admitted to our hospital form August 2012 and March 2015, were enrolled in our study. They were performed 1H-MRS before surgery, and apparently different regions of Cho/Cr were identified. With the help of intraoperative neuronavigation, different Cho/Cr tissue samples were gained accurately (Cho/Cr hypermetabolism group and Cho/Cr hypometabolism group). The different distribution of glioma stem cells in glioma tissues of the two groups was detected via neurosphere culture; immunohistochemistry and Western blotting were employed to detect the CD133 and nestin expressions. Results Neurospheres were successfully cultured from different glioma tissues, and the sphere formation rate from Cho/Cr hypermetabolism group was significantly higher as compared with that from Cho/Cr hypometabolism group (13.94±3.55 vs. 8.04± 1.47, P<0.05). The immunohistochemistry results indicated that the expressions of CD133 and nestin in the Cho/Cr hypermetabolism group were significantly higher as compared with those in the Cho/Cr hypometabolism group ([22.96±2.28]% vs. [18.04±1.36]%, [25.47±2.43]% vs. [19.74±1.66]%, P<0.05). Western blotting showed that the relative protein expressions of CD133 and nestin in the Cho/Cr hypermetabolism group were significantly higher as compared with those in the Cho/Cr hypometabolism group (0.50±0.17 vs. 0.30±0.08, 0.45±0.13 vs. 0.27±0.07, P<0.05); and the protein expressions of CD133 and nestin were positively correlated with Cho/Cr (r=0.972, P=0.000; r=0.762, P=0.000). Conclusion 1H-MRS parameter Cho/Cr reveals the distribution differences of cancer stem cells in high-grade gliomas, which can assist in finding and resecting the glioma stem cells-rich region.
2.Influences of ginsenoside Rg1 on blood-brain barrier damage and neuroinflam-mation in rats with experimental cerebral hemorrhage by regulating miR-144-3p/FPR2/p38 signaling pathway
Yalin BAI ; Zhanhai FANG ; Chenzhe DING ; Yanping LAN ; Dailin LIU ; Gaoyang QI ; Lei CHEN ; Juncheng WANG
Chinese Journal of Immunology 2023;39(12):2534-2539
Objective:To investigate influences of ginsenoside Rg1 regulating miR-144-3p on neuroinflammation and blood-brain barrier damage in rats with experimental cerebral hemorrhage,and its regulation on formyl peptide receptor 2(FPR2)/p38 path-way.Methods:Ninety SD rats were randomly divided into control group,cerebral hemorrhage group,ginsenoside Rg1 low-dose group(10 mg/kg),ginsenoside Rg1 high-dose group(40 mg/kg),ginsenoside Rg1 high-dose+ago-miR-144-3p group(40 mg/kg ginseno-side Rg1+ago-miR-144-3p),with 18 mice in each group.Except for control group,experimental intracerebral hemorrhage rat model was constructed by injecting collagenase Ⅱ into right caudate nucleus,and then each group was given intraperitoneal administration and intracerebral injection as required.Neurological damage in rats was scored;rat brain water content was determined by dry-wet spe-cific gravity method;levels of TNF-α,IL-6 and IL-1β in rat brain tissues homogenate were determined by ELISA;ultrastructure around cerebral edema was observed by electron microscope;permeability of blood-brain barrier in rats was determined by Evans blue(EB)method;expressions of miR-144-3p/FPR2/p38 pathway were determined by qRT-PCR and Western blot.Results:Compared with control group,blood-brain barrier damage was aggravated in cerebral hemorrhage group,neurological function damage score,brain water content,miR-144-3p,TNF-α,IL-6,IL-1β,p38 mRNA,p-p38/p38 expressions in brain homogenate were increased(P<0.05),FPR2 mRNA and protein expressions were decreased(P<0.05);compared with cerebral hemorrhage group,blood-brain barrier damage was reduced in ginsenoside Rg1 low-dose group and ginsenoside Rg1 high-dose group,neurological function damage score,brain water content,miR-144-3p,TNF-α,IL-6,IL-1β,p38 mRNA,p-p38/p38 expressions in brain homogenate were decreased(P<0.05),FPR2 mRNA and protein expressions were increased(P<0.05);ago-miR-144-3p was able to reverse protective effects of gin-senoside Rg1 on blood-brain barrier and neuroinflammation in rats(P<0.05).Conclusion:Ginsenoside Rg1 may inhibit blood-brain barrier damage and neuroinflammation in rats by regulating miR-144-3p/FPR2/p38 axis.
3.Effect of Modified Erchentang on Bronchioles of Rats with Chronic Obstructive Pulmonary Disease by HMGB1/RAGE/NF-κB Signaling Pathway
Lizhi SHANG ; Yaoyang LI ; Shu JI ; Wenying XIE ; Haofan SHANG ; Zhuang CHEN ; Gaoyang LIU ; Qi WANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(6):44-54
ObjectiveTo study the effect of modified Erchentang on the expression of key molecules in the high mobility group Box 1 protein (HMGB1)/receptor for advanced glycation endproduct (RAGE)/nuclear factor-κB (NF-κB) signaling pathway in bronchioles of rats with chronic obstructive pulmonary disease (COPD), to explore the mechanism of modified Erchentang against bronchiolar inflammation of COPD rats via HMGB1/RAGE/NF-κB signaling pathway. MethodSixty SD rats were randomly divided into normal group, model group, modified Erchentang low-, medium- and high-dose groups (5, 10, 20 g·kg-1·d-1) and ethyl pyruvate (HMGB1 inhibitor) group, with 10 in each group. The COPD rat model was prepared by cigarette smoke combined with tracheal injection of lipopolysaccharide (LPS). After modeling, the modified Erchentang groups were given corresponding drugs (ig) and Ringer's solution (4 mL, ip), while the EP group was treated with equal volume of normal saline (ig) and EP (0.04 g·kg-1·d-1, ip). The normal group and the model group received equal volume of normal saline (ig) and Ringer's solution (ip) for 21 consecutive days. The contents of HMGB1, chemokine (C-X-C motif) ligand 1 (CXCL1), CXCL2 and monocyte chemotactic protein-1 (MCP-1) in bronchoalveolar lavage fluid (BALF) were detected by enzyme-linked immunosorbent assay (ELISA). The mRNA expressions of HMGB1, RAGE and NF-κB p65 were determined by Real-time polymerase chain reaction (Real-time PCR), and the protein expressions of HMGB1, RAGE, p-NF-κB p65, and alpha-smooth muscle actin (α-SMA) in bronchioles tissue of rats were determined by immunohistochemistry (IHC). ResultCompared with the conditions in the normal group, the forced vital capacity (FVC), forced expiratory volume in the first second (FEV1) and FEV1/FVC in the model group were decreased (P<0.01) while the contents of HMGB1, CXCL1, CXCL2 and MCP-1 in BALF were increased (P<0.01). And the model group presented higher mRNA expressions of HMGB1, RAGE and NF-κB p65 (P<0.01) and protein expressions of HMGB1, RAGE, p-NF-κB p65 and α-SMA (P<0.05, P<0.01) than the normal group. Compared with the model group, the modified Erchentang medium- and high-dose groups had increased FEV1/FVC (P<0.05, P<0.01), lowered contents of HMGB1, CXCL1, CXCL2 and MCP-1 in BALF (P<0.05, P<0.05), and reduced mRNA expressions of HMGB1, RAGE and NF-κB p65 (P<0.05, P<0.01) and protein expressions of HMGB1, RAGE, p-NF-κB p65 and α-SMA (P<0.05, P<0.01). ConclusionModified Erchentang can resist bronchiolar inflammation of COPD rats. The mechanism may be related to down-regulating the mRNA expressiona of HMGB1 and RAGE, inhibiting the activity of NF-κB, and reducing the release of HMGB1, CXCL1, CXCL2 and MCP-1, thus suppressing the inflammatory injury and abnormal repair of bronchioles.
4.Optimization of the experimental condition on separating lipoproteins by self-constructed asymmetrical flow field-flow fractionation.
Di WU ; Bai Ling QIU ; Chen Qi ZHU ; Shuang GUO ; Ya Ya GAOYANG ; Qi Hui LIANG ; Ye GAO ; Yu SONG ; Nan Yin HAN
Journal of Peking University(Health Sciences) 2018;50(3):564-571
OBJECTIVE:
To analyze and characterize the separation effectiveness of self-constructed asymmetrical flow field-flow fractionation system on proteins and lipoproteins, to achieve the optimization of the experimental conditions when separating lipoproteins by orthogonal design test and to investigate the carrier viscosity's influence on separation effectiveness.
METHODS:
The evaluation of asymmetrical flow field-flow fractionation separation capacity was conducted by using two standard proteins (carbonic anhydrase and thyroglobulin). Under the optimized separation conditions of carbonic anhydrase and thyroglobulin, the channel actual thickness (after assembling, the actual thickness of separation channel was less than initial thickness) was calculated by the analytes' elution time based on the hydrokinetic theory. With orthogonal design test the optimized experimental conditions were studied and statistical analysis was carried on to find out the factors with statistical significance which needed further exploration.
RESULTS:
According to the hydrodynamics principle and Stoke's function, the channel actual thickness was measured to be 164 μm by separating the two standard proteins, carbonic anhydrase and thyroglobulin, under proper experimental conditions. By the optimization based on orthogonal design test, base-line separation (the resolution had to be higher than 1.50) was achieved. The resolutions of the two experiments were 1.61 and 1.58. According to previous study/ pre-study and supporting theory, in the orthogonal design test, the total 5 factors were integrated for comprehensive investigation: the total flow rate (3.00, 3.50, 4.00, 4.50 mL/min), focus time (3.00, 3.50, 4.00, 4.50 min), transition time (0.5, 1.0, 1.5, 2.0 min), pH of the carrier fluid(6.8, 7.00, 7.20, 7.40) and viscosity of the carrier fluid hydroxypropylmethylcellulose concentration: 0.00%, 0.03%, 0.06%, 1.00%). Among the 5 factors, viscosity was found to have the statistical significance on separation effectiveness which was further investigated. The resolution of high density lipoprotein and low density lipoprotein was increased by the increasing viscosity which also caused more obvious negative spikes.
CONCLUSION
The separating capacities of self-constructed asymmetrical flow field-flow fractionation system on lipoproteins were verified to be effective and an optimized experimental condition was found to achieve the base-line separation of high density lipoprotein and low density lipoprotein. Viscosity of the carrier fluid was proved to have the statistical significance on lipoprotein separation.
Fractionation, Field Flow
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Lipoproteins
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Lipoproteins, LDL