ObjectiveTo summarize the relationship between exosome and thyroid diseases.MethodThe literatures reports on exosomes and the physiology, pathology and diseases of thyroid were collected and reviewed.ResultsExosomes were secreted by cells and could be found in various body fluids, which could mediate the normal physiological development of the thyroid gland and play an important role in the progression of Graves’ disease. Exosomes could be used as diagnostic and differential diagnostic biomarkers for thyroid cancer and affect the growth, invasion, and metastasis of thyroid cancer. As a drug carrier for anti-thyroid cancer, exosome had a good targeting ability.ConclusionExosomes play an important role in the development of various diseases of the thyroid gland, which have good application prospects in biomarkers for early diagnosis and prognostic evaluation, as well as targeted drug carriers for thyroid cancer.
ObjectiveTo investigate the regulatory role of MSC-derived exosomes in obliterative bronchiolitis after lung transplantation. MethodsThe murine lung transplantation model was established with male C57BL/6 mice, and the mice were divided into a sham group (sham, n=6), a surgery group (OB, n=6), and a treatment group (OB+MSC-exo, n=6). The in vitro model was created by stimulating RAW264.7 with lipopolysaccharide+nigericin (LPS+Nigericin), and comprised a PBS group, a LPS+Nigericin group, and a LPS+Nigericin+MSC-exo group. Immunofluorescence and hematoxylin-eosin (HE) staining were used to analyze gasdermin D (GSDMD) expression, as well as lumen stenosis in lung grafts. Bioinformatics methods were employed to predict and screen target gene collagen type V alpha 1 (COL5A1). Q-PCR was used to measure mRNA expression levels of interleukin (IL)-1β, IL-18, IL-6, tumor necrosis factor-α (TNF-α), and COL5A1 in lung grafts and macrophages. Western blot was performed to detect Cleaved-Caspase 1 protein expression in lung grafts and GSDMD protein expression in macrophages. ResultsImmunofluorescence and HE staining revealed that in vivo infusion of MSC-exo reduced GSDMD expression in grafts, ameliorated tracheal epithelial cilia loss and lumen stenosis, and decreased Cleaved-Caspase 1 protein as well as IL-1β and IL-18 mRNA expression. MSC-exo treatment or COL5A1 knockdown reduced IL-1β, IL-18, IL-6, and TNF-α mRNA expression in macrophages, with comparable efficacy. MSC-exo infusion also decreased the number of COL5A1+ cells and mRNA expression levels in lung grafts. ConclusionMSC-derived exosomes alleviate obliterative bronchiolitis after lung transplantation by inhibiting COL5A1.
Objective To explore the effects of adipose-derived stem cell released exosomes (ADSC-Exos) on the proliferation, migration, and tube-like differentiation of human umbilical vein endothelial cells (HUVECs). Methods Adipose tissue voluntarily donated by liposuction patients was obtained. The ADSCs were harvested by enzyme digestion and identified by flow cytometry and adipogenic induction. The ADSC-Exos were extracted from the supernatant of the 3rd generation ADSCs and the morphology was observed by transmission electron microscopy. The surface proteins (Alix and CD63) were detected by Western blot. The nanoparticle tracking analyzer NanoSight was used to analyze the size distribution of ADSC-Exos. After co-culture of PKH26 fluorescently labeled ADSC-Exos with HUVECs, confocal microscopy had been used to observe whether ADSC-Exos could absorbed by HUVECs. ADSC-Exos and HUVECs were co-cultured for 1, 2, 3, 4, and 5 days. The effect of ADSC-Exos on the proliferation of HUVECs was detected by cell counting kit 8 (CCK-8) assay. The expression of VEGF protein in the supernatant of HUVECs with or without ADSC-Exos had been detected by ELISA after 12 hours. Transwell migration assay was used to detect the effect of ADSC-Exos on the migration ability of HUVECs. The effect of ADSC-Exos on the tubular structure formation of HUVECs was observed by Matrigel experiments in vitro. The formation of subcutaneous tubular structure in vivo was observed in BALB/c male nude mice via the injection of HUVECs and Matrigel with or without ADSC-Exos. After 2 weeks, the neovascularization in Matrigel was measured and mean blood vessel density (MVD) was calculated. The above experiments were all controlled by the same amount of PBS. Results After identification, the cultured cells were consistent with the characteristics of ADSCs. ADSC-Exos were circular or elliptical membranous vesicle with uniform morphology under transmission electron microscopy, and expresses the signature proteins Alix and CD63 with particle size ranging from 30 to 200 nm. Confocal microscopy results showed that ADSC-Exos could be absorbed by HUVECs. The CCK-8 analysis showed that the cell proliferation of the experimental group was better than that of the control group at each time point (P<0.05). The result of Transwell showed that the trans-membrane migration cells in the experimental group were significantly more than that in the control group (t=9.534, P=0.000). In vitro, Matrigel tube-forming experiment showed that the number of tube-like structures in the experimental group was significantly higher than that of the control group (t=15.910, P=0.000). In vivo, the MVD of the experimental group was significantly higher than that of the control group (t=16.710, P=0.000). The ELISA assay showed that the expression of VEGF protein in the supernatant of the experimental group was significantly higher than that of the control group (t=21.470, P=0.000). Conclusion ADSC-Exos can promote proliferation, migration, and tube-like structure formation of HUVECs, suggesting that ADSC-Exos can promote angiogenesisin vitro and in vivo.
Plant-derived exosome-like nanoparticle (PELN) is a nanoscale vesicle secreted by plant cells, which has important biological functions. On the one hand, PELN can exert anti-osteoporosis (OP) effects by affecting the functions of osteoclasts, osteoblasts, and bone marrow mesenchymal stem cells. On the other hand, PELN can also inhibit inflammatory reactions, protect chondrocytes, and has potential value in treating osteoarthritis (OA). This article summarizes the basic concepts, formation and components, separation and characterization methods of PELN, and focuses on the intervention effect and molecular mechanism of PELN on OP and OA.
ObjectiveTo summarize the research progress of mesenchymal stem cells derived exosomes (MSCs-EXOs) in wound repair in recent years.MethodsThe literature about the role of MSCs-EXOs in wound repair at home and abroad was extensively consulted. The mechanism of MSCs-EXOs in wound repair and its clinical application prospects were summarized and analyzed.ResultsMSCs-EXOs can inhibit early inflammatory reaction, promote angiogenesis, proliferation, and migration of epithelial cells, regulate collagen synthesis, and inhibit scar proliferation in the later stage of wound healing. Compared with MSCs, MSCs-EXOs have many advantages, such as high stability, easy storage, non-tumorigenicity, no proliferation, easy quantitative use, and so on. It has broad clinical application prospects.ConclusionMSCs-EXOs can promote wound repair and hopefully develop into a clinical product to promote the repair of acute or chronic wounds.
ObjectiveTo evaluate the effects of icariin on autophagy induced by low-concentration of glucocorticoid and exosome production in bone microvascular endothelial cells (BMECs).MethodsBMECs were isolated from femoral heads resected in total hip arthroplasty and then intervened with hydrocortisone of low concentration (0, 0.03, 0.06, 0.10 mg/mL), which were set as groups A, B, C, and D, respectively. On the basis of hydrocortisone intervention, 5×10?5 mol/L of icariin was added to each group (set as groups A1, B1, C1 and D1, respectively). Western blot was used to detect the expressions of microtubule-associated protein 1 light chain 3B (LC3B) and dead bone slice 1 (p62) after 24 hours. Exosomes were extracted from BMECs treated with icariin (intervention group) and without icariin (non-intervention group), and the diameter and concentration of exosomes were evaluated by nanoparticle tracking analysis technique. The total protein content of exosomes was detected by BCA method, and the expressions of proteins carried by exosomes including CD9, CD81, transforming growth factor β1 (TGF-β1), and vascular endothelial growth factor A (VEGFA) were assessed by Western blot. The BMECs were further divided into three groups: BMECs in the experimental group and the control group were co-cultured with exosomes secreted by BMECs treated with or without icariin, respectively; the blank control group was BMECs without exosome intervention. The three groups were treated with hydrocortisone and Western blot was used to detect the expressions of LC3B and p62. The scratching assay was used to detect cell migration ability; angiogenic ability of BMECs was also assessed.ResultsWith the increase of hydrocortisone concentration, the protein expression of LC3B-Ⅱ increased gradually, and the protein expression of p62 decreased gradually (P<0.01). Compared with group with same concentration of hydrocortisone, the protein expression of LC3B-Ⅱ decreased and the protein expression of p62 increased after the administration of icariin (P<0.01). The concentration of exosomes in the intervention group was significantly higher than that in the non-intervention group (t=?10.191, P=0.001); and there was no significant difference in exosome diameter and total protein content between the two groups (P>0.05). CD9 and CD81 proteins were highly expressed in the non-intervention group and the intervention group, and the relative expression ratios of VEGFA/CD9 and TGF-β1/CD9 proteins in the intervention group were significantly higher than those in the non-intervention group (P<0.01). After co-culture of exosomes, the protein expression of p62 increased in blank control group, control group, and experimental group, while the protein expression of LC3B-Ⅱ decreased. There were significant differences among groups (P<0.05). When treated with hydrocortisone for 12 and 24 hours, the scratch closure rate of the control group and experimental group was significantly higher than that of the blank control group (P<0.05), and the scratch closure rate of the experimental group was significantly higher than that of the control group (P<0.05). When treated with hydrocortisone for 4 and 8 hours, the number of lumens, number of sprouting vessels, and length of tubule branches in the experimental group and the control group were significantly greater than those in the blank control group (P<0.05); the length of tubule branches and the number of lumens in the experimental group were significantly greater than those in the control group (P<0.05).ConclusionIcariin and BMECs-derived exosomes can improve the autophagy of BMECs induced by low concentration of glucocorticoid.
Objective To explore whether blood exosome carrying miR-140-3p can regulate the malignant progression of small cell lung cancer (SCLC) through targeting ubiquitin-conjugating enzyme E2C (UBE2C). MethodsThis study was consisted of bioinformatics analysis, clinical research, cell analysis, and animal experiments. We searched GEO database for data of SCLC related microRNA (miRNA) dataset GSE19945, mRNA dataset GSE40275, and GSE60052. T-test was used to detect the differential expression of miR-140-3p in normal tissues and SCLC tissues in the dataset, and the expression of miR-140-3p in different tissues and extracellular vesicles was analyzed through a database. SCLC tissue and paired cancerous tissues excised at Yongzhou Central Hospital were collected between December 2021 and December 2022, and healthy volunteers 7 days before the start of the study was selected. Quantitative real-time polymerase chain reaction was used to detect the expression level distribution of miR-140-3p and UBE2C in tissue samples of SCLC patients and healthy volunteers. SCLC patients were divided into low expression and high expression groups based on the median expression level, and the correlation between the expression levels of miR-140-3p and UBE2C and patient pathological parameters was analyzed. 20 male nude mice was selected. The nude mice were randomly divided into 4 groups: miR-140-3p, UBE2C analog negative control group, and analog control group, with 5 mice in each group. Immunohistochemical detection system was used to detect tumor tissue sections in nude mice. Results A total of 45 patients and 30 healthy volunteers were included. SCLC malignant progression was significantly associated with the expression of miR-140-3p and UBE2C. The expression of miR-140-3p was low in blood-derived exosomes from SCLC patients. Overexpression of miR-140-3p inhibited the proliferation (47.33±2.52 vs. 107.67±10.69, P<0.05), migration [(11.63±2.62)% vs. (31.77±4.30)%, P<0.05] and invasion (44.33±3.06 vs. 102.67±8.50, P <0.05) and promoted their apoptosis [(14.48±1.20)% vs. (10.14±1.21)%, P<0.05]. Bioinformatics analysis yielded the target gene UBE2C of miR-140-3p. In vitro experiments further demonstrated that miR-140-3p directly targetd UBE2C to inhibit SCLC cell proliferation, migration, invasion, epithelial mesenchymal transition, and promote apoptosis. Mouse xenotransplantation experiments showed that miR-140-3p mimic significantly inhibited tumor growth. ConclusionTherefore, the miR-140-3p extracellular vesicle and the oncogenic gene UBE2C may be potential targets for inhibiting the malignant progression of SCLC.
ObjectiveTo review the advances in utilizing paracrine effect of stem cells in knee osteoarthritis (OA) treatment.MethodsThe researches in applying stem cells derived conditioned medium, extracellular matrix, exosomes, and microvesicles in knee OA treatment and cartilage repair were reviewed and analyzed.ResultsThe satisfying outcomes of using different products of stem cells paracrine effect in knee OA condition as well as cartilage defect is revealed in studies in vitro and in vivo. The mechanism including suppressing the intraarticular inflammation, the apoptosis of chondrocytes, and the degradation of cartilage matrix, while enhancing the synthesis of cartilage matrix, the differentiation of in-situ stem cells into chondrocytes and the migration to the affected area. The effectiveness can be further improved supplemented with the tissue engineering methods or gene modification.ConclusionCompared with the traditional stem cell therapy, applying the products from paracrine effect of stem cells in knee OA treatment is more economical and safer, presenting great potential in clinical practice.
ObjectiveTo summarize the molecular mechanisms and clinical treatment of gastric cancer with liver metastasis (GCLM), in order to provide new ideas for future treatment. MethodThe literatures about mechanism and treatment strategy of GCLM in recent years were searched and reviewed. ResultsMost patients with gastric cancer were in advanced stage or had developed distant metastases when they were first diagnosed, among which liver was the common site of metastasis. The complex molecular mechanisms of GCLM had not been fully clarified. Molecular mechanisms at different levels, including non-coding RNA, circulating tumor cells, exosomes, tumor microenvironment and signaling pathways, were relatively independent and interacted with each other, providing potential biomarkers and therapeutic targets for GCLM. At present, the best treatment method for patients with GCLM was mainly divided into local and systemic treatment. The local treatment included surgical treatment, radiofrequency ablation and proton beam therapy, while the systemic treatment included systemic chemotherapy, targeted therapy and immunotherapy, among which the targeted therapy and immunotherapy were the focus of recent research. ConclusionsThe mechanism of GCLM is the result of the interaction between tumor cells and the microenvironment at the site of metastasis. Understanding them is of great significance to guide clinical treatment and prognosis. At present, there is no unified treatment standard for GCLM. To achieve the ideal treatment effect, we should not only rely on single therapy, but also adopt multi-disciplinary and individual therapy according to the specific disease status of patients and the nature of tumors.
ObjectiveTo understand the research progress of related biomarkers in early diagnosis of gastric cancer in recent years.MethodThe domestic and foregin literatures on studies of biomarkers of early diagnosis of gastric cancer in recent years were reviewed.ResultsAt present, the sensitivity and specificity of serum tumor biomarkers of gastric cancer such as CEA and CA19-9 were lower, so the molecular markers that could predict, screen, and diagnose gastric cancer in the early stage were further explored. The recent studies suggested that microRNAs, long non-coding RNAs, circular RNAs, exosome, etc. molecular markers in early diagnosis of gastric cancer had better prospects of clinilal application.ConclusionWith the continuous development of molecular biology technology, the values of microRNAs, long non-coding RNAs, circular RNAs, DNA, etc. in early diagnosis of gastric cancer would be further explored.
