Apocynum venetum leaf extract alleviated doxorubicin-induced cardiotoxicity by regulating organic acid metabolism in gut microbiota.

Apocynum venetum leaf is commonly utilized for its beneficial effects in reducing blood pressure, inducing sedation, promoting diuresis, anti-aging, and cardioprotection, which also exhibit positive effects on the gut microbiota. The gut microbiota plays a role as an endocrine organ by producing bioactive metabolites that can directly or indirectly impact host physiology, specifically cardiovascular diseases. In this study, main chemical components of A. venetum leaf extract (AVLE) were identified by LC-MS, and an orally administered AVLE was employed to treat mice with doxorubicin (Dox)-induced cardiotoxicity. The results showed that AVLE contained hyperoside and oganic acids. The pharmacological findings revealed that AVLE regulated the gut microbiota, resulting in a significant increase in the levels of two organic acids, indole-3-propionic acid (IPA) and acetic acid (AA). Both IPA and AA exhibited the ability to reduce BNP, CK, and LDH levels in mice with Dox-induced cardiotoxicity. Moreover, IPA demonstrated an improvement in Dox-induced cardiac injury by inhibiting apoptosis, while AA promoted increased secretion of ghrelin through the parasympathetic nervous system, subsequently reducing cardiac fibrosis by decreasing collagen I, collagen III, and activin A. Hence, our study demonstrates that AVLE exerts a beneficial cardioprotective effect by modulating the gut microbiota, providing a potential novel target for the treatment and prevention of Dox-induced cardiotoxicity.

Progress and obstacles in transplantation of brown adipose tissue or engineered cells with thermogenic potential for metabolic benefits.

Transplantation of brown adipose tissue (BAT), engineered thermogenic progenitor cells, and adipocytes have received much attention for the improvement of obesity and metabolic disorders. However, even though the thermogenic and metabolic potential exists early after transplantation, the whitening of the brown fat graft occurs with metabolic function significantly impaired. In this review, specific experiment designs, graft outcomes, and metabolic benefits for the transplantation of BAT or engineered cells will be discussed. The current advancements will offer guidance to further investigation, and the obstacles appearing in previous studies will require innovation of BAT transplantation methods.

Condensing of Low-Density Fat by Mechanical Process Improves Fat Retention and Reduces Oil Cyst Formation in Breast Reconstruction.

BACKGROUND: Although autologous fat grafting is a useful adjunct for breast reconstruction, its indications remain limited as large-volume fat grafting results in high absorption and complication rates. Low-density fat includes small numbers of viable cells and considerable oil, resulting in nodules and oil cysts. This study evaluated the volumization effect and complications with combined fat grafting of condense low-density fat and high-density fat. METHODS: This retrospective, single-center study included 25 patients who underwent combined grafting of condensed low-density fat and high-density fat (CLDF + HDF) and 20 patients who underwent conventional Coleman fat grafting for breast reconstruction from December 2017 to January 2022. Retention rates and complications were evaluated by magnetic resonance imaging and ultrasound rates. Patient satisfaction was evaluated using a typical Likert scale. Photographs were taken and imageological examinations were performed before and after treatment. OUTCOMES: Graft retention rate was higher in patients who underwent CLDF + HDF than Coleman fat grafting for breast reconstruction (38.40 ± 4.41% vs. 31.43 ± 5.43%, p <0.05). One patient in the CLDF + HDF grafting group, compared with twelve in the Coleman fat grafting group, developed oil cysts exceeding 1 cm. Patient satisfaction rate was higher in the CLDF + HDF grafting group. CONCLUSIONS: Mechanical processes can concentrate the cellular content of LDF and remove oil, condensing LDF to the level of HDF. Combined grafting of CLDF optimized by mechanical processing and HDF is effective for breast reconstruction, with a higher retention rate and a lower incidence of complications than conventional Coleman fat grafting. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

The fates of different types of adipose tissue after transplantation in mice.

Fat grafting is one of the most commonly applied procedure for soft-tissue repair. However, it remains unclear whether the type of adipose tissue would have any effects on fat graft survival. The present study aimed to determine fates of fat grafting of three different types of fat tissue. In this study, mice were randomly divided into three groups, white adipose tissue (WAT) group, beige adipose tissue (beige AT) group and brown adipose tissue (BAT) group. Before transplantation, donor mice were injected with rosiglitazone or phosphate-buffered saline (PBS). The WAT and BAT were obtained from PBS-treated mice while beige AT was obtained from the rosiglitazone-treated mice. Three types of fat tissue (150 mg each) were transplanted in three groups, respectively, and harvested at 2, 4 or 12 weeks. The BAT and beige AT contained smaller adipocytes and expressed higher level of uncoupling protein-1 gene. The retention rate of the transplanted fat was significantly higher for beige than for white fat, but was significantly lower for brown than for white fat. Transplanted brown fat was characterized by upregulated inflammation and high endoplasmic reticulum stress. By contrast, fat grafts in beige AT group showed the best adipogenic capacity, moderate inflammation level and superior angiongenesis. In vitro, under hypoxic condition, fewer apoptotic cells were found in beige adipocyte group than that in brown and white adipocyte group. Conditioned medium from brown adipocytes induced M1 polarization of RAW 264.7 macrophages while that from beige adipocytes effectively promoted M2 polarization. Therefore, we suggest that beige AT provides a new potential choice for fat grafting because of low inflammation and superior survival but BAT might not be ideal for fat grafting due to its poor survival.

Adipose-Derived Stem Cell-Enriched Lipotransfer Reverses Skin Sclerosis by Suppressing Dermal Inflammation.

BACKGROUND: Scleroderma is a chronic autoimmune disease with an incidence of 2.7 per 100,000 people. Traditional lipotransfer has been used to treat atrophic sclerotic skin. Enzymatically processed cell-assisted lipotransfer and mechanically processed stromal vascular fraction gel are fat products with abundant adipose-derived stem cells. This study assessed whether adipose-derived stem cell-enriched lipotransfer elicits superior therapeutic effects on scleroderma. METHODS: Scleroderma was induced in nude mice by injections of bleomycin for 4 weeks. Human-derived Coleman fat, cell-assisted lipotransfer, or stromal vascular fraction gel (0.1 ml) was injected into sclerotic lesions. Histologic examinations, terminal deoxynucleotidyl transferase dUTP nick end labeling, and expression analyses of inflammatory factors in skin lesions and transferred fat were performed at 4 weeks after implantation. RESULTS: Dermal thickness was lower in the groups injected with Coleman fat (339.0 ± 19.66 µm), cell-assisted lipotransfer (271.0 ± 16.15 µm), and stromal vascular fraction gel (197.8 ± 12.99 µm) than in the group injected with phosphate-buffered saline (493.3 ± 28.13 µm) ( p < 0.05). The numbers of terminal deoxynucleotidyl transferase dUTP nick end labeling + and Mac2 + cells in fat tissue were significantly higher in the group injected with Coleman fat than in those injected with stromal vascular fraction gel and cell-assisted lipotransfer. Expression of monocyte chemotactic protein-1 and interleukin-6 was significantly lower in the adipose-derived stem cell-enriched groups than in the Coleman fat group. Histologic analysis showed there were far fewer macrophages and myofibroblasts in skin lesions in the adipose-derived stem cell-enriched groups than in the Coleman fat group. CONCLUSIONS: Transplantation of stromal vascular fraction gel and cell-assisted lipotransfer, which contain abundant adipose-derived stem cells, reduces the levels of apoptotic cells and inflammation, significantly reverses skin sclerosis, and elicits superior anti-inflammatory and antifibrotic effects on scleroderma. CLINICAL RELEVANCE STATEMENT: This study provided an alternative adipose-based therapy, adipose-derived stem cell-enriched fat, for sclerotic lesions and showed its validity for interfering with the inflammation and fibrosis.

Skeletal muscle provides a pro-browning microenvironment for transplanted brown adipose tissue to maintain its effect to ameliorate obesity in ob/ob mice.

Brown adipose tissue (BAT) transplantation is a promising means of increasing whole-body energy metabolism to ameliorate obesity. However, the changes in BAT following transplantation and the effects of the microenvironment of the recipient site on graft function have yet to be fully characterized. Therefore, we aimed to determine the effects of transplanting BAT from C57BL/6 mice into the dorsal subcutaneous region or deep to the quadriceps femoris muscle of leptin-deficient ob/ob mice. Subcutaneously transplanted BAT lost features of BAT and demonstrated greater inflammatory cell infiltration and more oil cysts 16 weeks following transplantation. By contrast, the sub-muscularly transplanted BAT maintained features of BAT and was more highly vascularized. Interestingly, sub-muscular BAT transplantation led to a significant increase in oxygen consumption and less inflammation in subcutaneous fat, which was associated with long-term reductions in insulin resistance and body mass gain, whereas the subcutaneous transplants failed after 16 weeks. These results demonstrate that the beneficial effects of BAT transplantation depend upon the microenvironment of the recipient site. Skeletal muscle may provide a microenvironment that maintains the inherent features of BAT grafts over a long period of time, which facilitates a reduction in obesity and improvements in glucose homeostasis.

Improving Low-Density Fat by Condensing Cellular and Collagen Content through a Mechanical Process: Basic Research and Clinical Applications.

BACKGROUND: Large-volume fat grafting results in high absorption and complication rates. Low-density fat includes small numbers of viable cells and considerable oil, resulting in nodules and oil cysts. This study evaluated a strategy for large-volume fat grafting using a mechanical process to condense low-density fat and transplanting it with high-density fat. METHODS: Low-density fat, defined as the upper half of centrifuged lipoaspirates, was emulsified by intersyringe shifting and centrifuged to obtain condensed low-density fat. Fresh condensed low-density fat was analyzed by counting cells in the stromal vascular fraction, and by electron scanning and Western blotting. The retention rate and histologic changes of the product were analyzed using a fat grafting model in nude mice. Transplantation with a combination of condensed low-density fat and high-density fat was tested in patients undergoing breast reconstruction and breast augmentation. RESULTS: The condensed low-density fat derived from low-density fat contained a large number of stromal vascular fraction cells and collagens, comparable to that of high-density fat and much higher than in low-density fat and Coleman fat. Retention rates 12 weeks after transplantation were higher for condensed low-density fat (55.0 ± 7.5 percent) than for low-density fat (31.1 ± 5.7 percent) and Coleman fat (41.1 ± 6.8 percent), with condensed low-density fat having fewer oil cysts and lower macrophage infiltration. Patients grafted with combined condensed low-density fat and high-density fat showed good long-term volume retention. CONCLUSIONS: Using mechanical methods to condense low-density fat to a level comparable to that of high-density fat is a practical method of improving fat graft retention and avoiding severe complications. This new strategy may improve the quality of lipoaspirates for patients requiring large-volume augmentation.

Conditioned medium from primary cytotrophoblasts, primary placenta-derived mesenchymal stem cells, or sub-cultured placental tissue promoted HUVEC angiogenesis in vitro.

BACKGROUND: As a large capillary network, the human placenta plays an important role throughout pregnancy. Placental vascular development is complex and delicate and involves many types of placental cells, such as trophoblasts, and mesenchymal stem cells. There has been no systematic, comparative study on the roles of these two groups of placental cells and the whole placental tissue in the placental angiogenesis. In this study, primary cytotrophoblasts (CTBs) from early pregnancy and primary human placenta-derived mesenchymal stem cells (hPDMSCs) from different stages of pregnancy were selected as the cell research objects, and full-term placental tissue was selected as the tissue research object to detect the effects of their conditioned medium (CM) on human umbilical vein endothelial cell (HUVEC) angiogenesis. METHODS: We successfully isolated primary hPDMSCs and CTBs, collected CM from these placental cells and sub-cultured placental tissue, and then evaluated the effects of the CM on a series of angiogenic processes in HUVECs in vitro. Furthermore, we measured the levels of angiogenic factors in the CM of placental cells or tissue by an angiogenesis antibody array. RESULTS: The results showed that not only placental cells but also sub-cultured placental tissue, to some extent, promoted HUVEC angiogenesis in vitro by promoting proliferation, adhesion, migration, invasion, and tube formation. We also found that primary placental cells in early pregnancy, whether CTBs or hPDMSCs, played more significant roles than those in full-term pregnancy. Placental cell-derived CM collected at 24 h or 48 h had the best effect, and sub-cultured placental tissue-derived CM collected at 7 days had the best effect among all the different time points. The semiquantitative angiogenesis antibody array showed that 18 of the 43 angiogenic factors had obvious spots in placental cell-derived CM or sub-cultured placental tissue-derived CM, and the levels of 5 factors (including CXCL-5, GRO, IL-6, IL-8, and MCP-1) were the highest in sub-cultured placental tissue-derived CM. CONCLUSIONS: CM obtained from placental cells (primary CTBs or hPDMSCs) or sub-cultured placental tissue contained proangiogenic factors and promoted HUVEC angiogenesis in vitro. Therefore, our research is helpful to better understand placental angiogenesis regulation and provides theoretical support for the clinical application of placental components, especially sub-cultured placental tissue-derived CM, in vascular tissue engineering and clinical treatments.

Downregulation of Siah1 promotes colorectal cancer cell proliferation and migration by regulating AKT and YAP ubiquitylation and proteasome degradation.

BACKGROUND: Colorectal cancer (CRC) is one of the most common malignant tumors in the world. Siah E3 ubiquitin protein ligase 1 (Siah1) has been identified as a tumor suppressor gene and plays an important role in the development of malignant tumors. However, the potential role and molecular mechanism of Siah1 in the development and progression of CRC is still unclear. METHODS: To explore the role and molecular mechanism of Siah1 in the development and progression of CRC, we examined the expression of Siah1 in CRC tissue samples and analyzed its association with progression and prognosis in CRC. In addition, overexpression and knockdown of Siah1 was used to investigate its activity in CRC cells. We also use bioinformatics to analyze and verify the significant roles of Siah1 in critical signaling pathways of CRC. RESULTS: We found that the expression of Siah1 was significantly downregulated in CRC tissues, and low expression of Siah1 was associated with aggressive TNM staging and poor survival of CRC patients. Moreover, we revealed that overexpression of Siah1 in CRC cells markedly inhibited CRC cell proliferation and invasion in vitro and in vivo, while knockdown of Siah1 enhanced CRC cell proliferation and invasion. Furthermore, we found that Siah1 prohibited cell proliferation and invasion in CRC partially through promoting AKT (the serine-threonine protein kinase) and YAP (yes associated protein) ubiquitylation and proteasome degradation to regulate the activity of MAPK(mitogen-activated protein kinase 1), PI3K-AKT (phosphatidylinositol 3-kinase-the serine-threonine protein kinase) and Hippo signaling pathways. CONCLUSIONS: These findings suggested that Siah1 is a novel potential prognostic biomarker and plays a tumor suppressor role in the development and progression of CRC.

Fat Grafting for Facial Rejuvenation Using Stromal Vascular Fraction Gel Injection.

This study used stromal vascular fraction gel (SVF-gel), a mechanically processed fat-derived product, to treat eye bag and tear trough deformity. SVF-gel is prepared by a process of centrifugation and intersyringe shifting and is particularly rich in SVF cells and native adipose extracellular matrix. SVF-gel injection is used alone or combined with transconjunctival eye bag removal. High satisfaction was noted among patients treated with SVF-gel injection for periorbital rejuvenation with fairly low complication rates. SVF-gel injection is a good alternative to assist transconjunctival lower eyelid blepharoplasty and correct the palpebromalar groove, tear trough deformity, and supraorbital hollow.

TLE3 represses colorectal cancer proliferation by inhibiting MAPK and AKT signaling pathways.

BACKGROUND: Transducin-like enhancer of Split3 (TLE3) serves as a transcriptional corepressor during cell differentiation and shows multiple roles in different kinds of cancers. Recently, TLE3 together with many other genes involved in Wnt/ß-catenin pathway were detected hyper-methylated in colorectal cancer (CRC). However, the potential role and the underlying mechanism of TLE3 in CRC progression remain scarce. METHODS: Gene expression profiles were analyzed in The Cancer Genome Atlas (TCGA) microarray dataset of 41 normal colorectal intestine tissues and 465 CRC tissues. Western blot and Real-time Quantitative PCR (RT-qPCR) were respectively performed to detect protein and mRNA expression in 8 pairs of CRC tissue and matched adjacent normal mucosa. Immunohistochemistry (IHC) was conducted to evaluate TLE3 protein expression in 105 paraffin-embedded, archived human CRC tissues from patients, whose survival data were analyzed with Kaplan-Meier method. In vitro experiments including MTT assay, colony formation assay, and soft agar formation assay were used to investigate the effects of TLE3 on CRC cell growth and proliferation. Additionally, subcutaneous tumorigenesis assay was performed in nude mice to confirm the effects of TLE3 in vivo. Furthermore, gene set enrichment analysis (GSEA) was run to explore potential mechanism of TLE3 in CRC, and then we measured the distribution of CRC cell cycle phases and apoptosis by flow cytometry, as well as the impacts of TLE3 on MAPK and AKT signaling pathways by Western blot and RT-qPCR. RESULTS: TLE3 was significantly down-regulated in 465 CRC tissues compared with 41 normal tissues. Both protein and mRNA expressions of TLE3 were down-regulated in CRC compared with matched adjacent normal mucosa. Lower expression of TLE3 was significantly associated with poorer survival of patients with CRC. Besides, knock down of TLE3 promoted CRC cell growth and proliferation, while overexpression of TLE3 showed suppressive effects. Furthermore, overexpression of TLE3 caused G1-S phase transition arrest, inhibition of MAPK and AKT pathways, and up-regulation of p21Cip1/WAF1 and p27Kip1. CONCLUSION: This study indicated that TLE3 repressed CRC proliferation partly through inhibition of MAPK and AKT signaling pathways, suggesting the possibility of TLE3 as a biomarker for CRC prognosis.