Artemisinin derivatives inhibit adipogenic differentiation of 3T3-L1 preadipocytes through upregulation of CHOP.

Artemisinin derivatives could inhibit adipogenic differentiation of 3T3-L1 preadipocytes and prevent obesity in mice. However, the molecular mechanism remains largely unclear. Our research was designed to investigate the specific molecular target of artemisinin derivatives in adipogenic differentiation of 3T3-L1 preadipocytes. Here, we revealed that in response to dihydroartemisinin (DHA) or artesunate (ATS), intracellular lipid was decreased in a concentration dependent manner as shown by BODIPY staining. Quantitative PCR analysis showed that expression of Cebpa, Pparg, Fabp4 and Plin was significantly decreased by DHA treatment in a concentration and time dependent manner. Also, DHA treatment remarkably downregulated expression of CCAAT/enhancer-binding protein α (C/EBPα) and nuclear receptor peroxisome proliferation-activated receptor γ (PPARγ) of adipogenic induced 3T3-L1 cells as assayed by western blotting. RNA-seq analysis identified thousands of differential expression genes (DEGs), among which CHOP expression was significantly improved in DHA treated cells. Upregulation of CHOP was verified by quantitative PCR and western blotting, respectively. Knockdown of CHOP by the specific shRNA revealed that the inhibition of adipogenesis by DHA was strongly blocked, resulting in restored lipid accumulation and expression of adipogenic molecules. In conclusions, the inhibitory effect of DHA on adipogenic differentiation of 3T3-L1 preadipocytes was exerted in a concentration and time dependent manner, which was mediated by expression of CHOP.

Protective effect of microRNA-134-3p on multiple sclerosis through inhibiting PRSS57 and promotion of CD34+ cell proliferation in rats.

MicroRNAs (miRs) have been extensively studied for their involvement in multiple sclerosis (MS). We investigated the involvement of miR-134-3p on MS. The MS rat model was established, and positive expression of interleukin-17 (IL-17) was detected using the immunohistochemical method while the expression of miR-134-3p and serine protease 57 (PRSS57) was determined by means of reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Western blot analysis. Second, the miR-134-3p overexpression or short hairpin RNA against PRSS57 was introduced into the CD34+ cells to investigate the levels of proliferation and apoptosis-related genes by RT-qPCR and Western blot analysis. In addition, analysis of the targeting relations of miR-134-3p and PRSS57 was conducted using online software and dual-luciferase reporter gene assay. Furthermore, neuronal functions, inflammatory response, proliferation, and apoptosis of CD34+ cells were assayed by flow cytometry, enzyme-linked immunosorbent assay, and methyl thiazolyl tetrazolium. IL-17 and PRSS57 expression increased while miR-134-3p expression decreased in the spinal cord from MS rats. miR-134-3p could target PRSS57. miR-134-3p overexpression or PRSS57 silencing enhanced mitochondrial activity of neurons, mitochondrial membrane potential content, CD34+ cell proliferation, while decreasing Cyt C content, inflammatory response, and cell apoptosis. Collectively, overexpression of miR-134-3p promotes CD34+ cell proliferation via inhibition of PRSS57 in MS, which may serve as a promising target for MS intervention.

Phosphorylated Alpha-Synuclein in Red Blood Cells as a Potential Diagnostic Biomarker for Multiple System Atrophy: A Pilot Study.

Diagnosis of multiple system atrophy (MSA) remains a challenge, due to the complexity and overlapping of its symptoms with other Parkinsonian disorders. The critical role of alpha-synuclein (α-syn) in the pathogenesis of MSA makes it an ideal biomarker for the diagnosis of MSA. Although α-syn alterations in cerebrospinal fluid (CSF) and blood plasma have been extensively assessed for the utility in diagnosing MSA, inconsistent results have been obtained, presumably due to the contamination by hemolysis and other confounding factors. In this study, levels of serine 129-phosphorylated α-syn (pS-α-syn), a major pathologic form of α-syn, in red blood cells (RBCs), were measured using ELISA in a Chinese cohort consisting of 107 MSA patients and 220 healthy controls. A significant increase in the levels of pS-α-syn in RBCs (pS-α-syn-RBC) was observed in MSA patients than in healthy controls (14.02 ± 4.02 ng/mg versus 11.89 ± 3.57 ng/mg; p < 0.0001). Receiver operating characteristic curve (ROC) indicated that pS-α-syn-RBC discriminated the patients well from the controls with a sensitivity of 80.37% (95% confidence interval (CI): 71.58%-87.42%), a specificity of 88.64% (95% CI: 83.68%-92.51%), and an area under the curve (AUC) of 0.91 (95% CI: 0.87-0.94). The levels of pS-α-syn-RBC were negatively correlated with RBD-HK scores and differed between MSA-P and MSA-C subtypes (13.27 ± 1.91 versus 12.19 ± 3.04; p=0.025). The difference between subtypes was seen at Hoehn and Yahr stages 3 and 4, and the age at onset (AAO) between 60 and 69 years (p=0.016). The results suggest that pS-α-syn-RBC is increased in MSA patients and can be used as a potential diagnostic biomarker for MSA.

A cohort study of isolated brainstem infarction based on head MR imaging and clinical findings.

OBJECTIVE: The prognosis of patients with isolated brainstem infarction (BSI) differs on an individual patient basis. This study was undertaken to analyze the influences of different imaging and clinical features with the prognosis of patients with BSI. METHODS: The study population was derived from a multicenter study of intracranial atherosclerosis in China. In the present study, 300 patients were selected who had experienced non-cardiogenic brain stem infarction within the prior 7 days. Evaluations included clinical characteristics, location and size of the brainstem infarction, and whether the infarction was located in multiple perforating branches of the brainstem. Poor prognosis was defined as the presence of disability within 1 year from the onset of disease. RESULTS: In total, 281 patients were followed up at 1 year post-infarction. Of these 281 patients, 84 (29.9%) exhibited disability at 1 year; these patients showed a median National Institutes of Health Stroke Scale score of 6 on admission. Multiple logistic regression analysis showed that patients with BSI located in the territory of multiple perforating arteries, who were discharged without administration of statins, showed a poor 1-year prognosis. CONCLUSION: Isolated BSI involving multiple perforating arteries, without statin medication at discharge, indicated poor prognosis for patients with BSI.