ABSTRACT
Nerve regeneration in adult mammalian spinal cord is poor because of the lack of intrinsic regeneration of neurons and extrinsic factors - the glial scar is triggered by injury and inhibits or promotes regeneration. Recent technological advances in spatial transcriptomics (ST) provide a unique opportunity to decipher most genes systematically throughout scar formation, which remains poorly understood. Here, we first constructed the tissue-wide gene expression patterns of mouse spinal cords over the course of scar formation using ST after spinal cord injury from 32 samples. Locally, we profiled gene expression gradients from the leading edge to the core of the scar areas to further understand the scar microenvironment, such as neurotransmitter disorders, activation of the pro-inflammatory response, neurotoxic saturated lipids, angiogenesis, obstructed axon extension, and extracellular structure re-organization. In addition, we described 21 cell transcriptional states during scar formation and delineated the origins, functional diversity, and possible trajectories of subpopulations of fibroblasts, glia, and immune cells. Specifically, we found some regulators in special cell types, such as Thbs1 and Col1a2 in macrophages, CD36 and Postn in fibroblasts, Plxnb2 and Nxpe3 in microglia, Clu in astrocytes, and CD74 in oligodendrocytes. Furthermore, salvianolic acid B, a blood-brain barrier permeation and CD36 inhibitor, was administered after surgery and found to remedy fibrosis. Subsequently, we described the extent of the scar boundary and profiled the bidirectional ligand-receptor interactions at the neighboring cluster boundary, contributing to maintain scar architecture during gliosis and fibrosis, and found that GPR37L1_PSAP, and GPR37_PSAP were the most significant gene-pairs among microglia, fibroblasts, and astrocytes. Last, we quantified the fraction of scar-resident cells and proposed four possible phases of scar formation: macrophage infiltration, proliferation and differentiation of scar-resident cells, scar emergence, and scar stationary. Together, these profiles delineated the spatial heterogeneity of the scar, confirmed the previous concepts about scar architecture, provided some new clues for scar formation, and served as a valuable resource for the treatment of central nervous system injury.
Subject(s)
Mice , Animals , Gliosis/pathology , Cicatrix/pathology , Spinal Cord Injuries , Astrocytes/metabolism , Spinal Cord/pathology , Fibrosis , Mammals , Receptors, G-Protein-CoupledABSTRACT
Objective To investigate the expression of miRNA-16 in peripheral blood monouclear cells (PBMC)from systemic lupus erythematosus( SLE)patients. Methods Sixteen SLE patients who meet the diagnostic criteria of SLE revised in 1997 American rheumatology and 12 healthy individuals were selected as our subjects. Their peripheral blood were sampled. Total RNAs were extracted and purified. The level of miRNA-16 was determined by quantitative reverse transcription PCR( qRT-PCR). U6 was used as housekeeping control. The amount of target miRNA was normalized relative to the amount of U6(ΔCt =ΔCt miRNA-ΔCtU6 ). Relative expression levels were expressed as 2-ΔCt . Results The expression level of miRNA-16 in the SLE patients was 919. 87 ± 715. 45,significantly higher than that in the healthy control group(413. 6 3 ± 330. 69;t= -2. 497,P﹤0. 05). And miRNA-16 expression in SLE active group was 1 298. 79 ± 803. 79,significantly higher than that in SLE stable group(540. 95 ± 350. 15;t= -2. 445,P﹤0. 05). The level of miRNA-16 was related with AnuA (r=0. 669,P=0. 005),ESR(r=0. 608,P=0. 012)and SLEDAI(r=0. 530,P=0. 035). Conclusion The expression of miRNA-16 is high in SLE patients and it is related with SLE activity.
ABSTRACT
ObjectiveTo investigate the expression of miR-155 and miR-146a in peripheral blood mononuclear cells (PBMC) and plasma of rheumatoid arthritis (RA) patients.MethodsPBMC and plasma were separated from the peripheral blood of 34 RA patients and 15 healthy individuals.Total RNAs were isolated and miRNAs were purified.The levels of miR-155 and miR-146a were determined by quantitative reverse transcription PCR(qRT-PCR).U6 was used as housekeeping control.The amount of target miRNA was normalized relative to the amount of U6(ΔCt=ΔCtmiRNA-ΔCtU6).Relative expression levels were expressed as 2 △-ΔCt.Data were analyzed using SPSS 13.0 software.The test of homogeneity of variance and unpaired t-test was used to compare between groups.P values(2-tailed) less than 0.05 were considered as statistically significant.ResultsThe expressions of PBMC and plasma miR-155 were higher in RA patients than those in the healthy control individuals(0.08±0.08 vs 0.05±0.03,t=-2.225,P＜0.05; 5.9±6.7 vs 1.3±2.0,t=-3.677,P＜0.05).The expression of miR-146a in PBMC and plasma of RA patients and controls were(1.3±1.2 vs 0.8±0.6,t=-2.154,P＜0.05)and(741±1001 vs 300±295,t=-1.669,P＞0.05).According to their DAS28 value,RA patients were divided into high activity group (23 cases,DAS28≥5.0) and low disease activity group( 11cases,DAS28＜5.0).The plasma miR-155 and miR-146a expressions were significantly higher in high activity group than those in low activity group.There were no significant differences in the expression of PBMC miR-155 and miR-146a between the two groups.ConclusionThe expression of PBMC and plasma miR-155 and miR-146a are higher in RA patients.The expression of plasma miR-155 and miR-146a are associated with RA patients' activity.Plasma miR-155 and miR-146a may be potential non-invasive biomarkers for RA diagnosis anddisease activity assessment.
ABSTRACT
Aim To study the preventive and therapeutic effects of Buyang Huanwu Decoction(BYHWD)on denervated tibial muscle atrophy of rats.Methods After 60 Sprague-Dawley rats were subjected to Common Peroneal nerve crush model of 5 mm injury, they were randomly divided into 6 groups for daily intragastric administration of drugs:BYHWD high-dose, medium-dose, low-dose groups, mecobalamin group(positive control), model group, sham operation group.The drug administration lasted for 18 days.18 days after operation, slice, masson staining and analysis morphology were performed. The wet weight ratio and section area of tibial muscle were also measured.Results Tibial muscle section area of sham operation group was large, morphous was regular, while that of model group significantly diminished, structure was chaotic, and connective tissue hyperplasia was more obvious;Tibial muscle section area of BYHWD group and mecobalamin group was fairly diminished, morphous was fairly regular, and connective tissue hyperplasia was not obvious.Compared with model group, the section area of BYHWD group all increased significantly(P <0.01)in a dose-dependent manner.Compared with mecobalamin group, the cross section area of BYHWD high-dose group was significantly larger(P <0.05).Compared with model group, the wet weight ratio of BYHWD high-dose and medium-dose group all increased significantly(P <0.01 or P <0.05)in a dose-dependent manner.Conclusion BYHWD has a significant effect of prevention and therapy on denervated tibial muscle atrophy of rats.