Mini Review - Stem Cell Research and Regenerative Medicine (2023) Volume 6, Issue 2
Promotes a coronary in Acute Cardiovascular Cells via Modulating Transcription
Department of Stem Cell and Research, India
Department of Stem Cell and Research, India
Received: 01-Apr-2023, Manuscript No. srrm-23-95867; Editor assigned: 04-Apr-2023, Pre-QC No. srrm-23- 95867 (PQ); Reviewed: 18-Apr-2023, QC No. srrm-23-95867; Revised: 22- Apr-2023, Manuscript No. srrm-23- 95867 (R); Published: 28-Apr-2023, DOI: 10.37532/srrm.2023.6(2).24-27
Diabetes mellitus has a higher incidence of cardiovascular disease, including Impaired microcirculation in the lower extremities where angiogenic disturbances are the main factor. The endothelium acts as a barrier between the blood and the vascular wall. Dysfunction of vascular endothelial cells by hyperglycemia is a major factor leading to impaired angiogenesis. Hydrogen sulfide (H2S) and miR-126-3p are known for their pro-angiogenic effects. However, little is known about how H2S regulates miR-126-3p to promote angiogenesis under high glucose conditions. The primary aim of this study was to investigate how H2S regulates miR-126-3p levels under high glucose conditions. We evaluated the pro-angiogenic effect of H2S in the diabetic hindlimb of an ischemic mouse model and in vivo Matrigel plugs. Using two microRNA datasets, we searched for microRNAs regulated by both diabetes and H2S. mRNA and protein levels were detected by real-time PCR and western blot, respectively. Immunofluorescence staining has also been used to determine capillary density and assess protein levels in vascular endothelial cells. We demonstrated endothelial cell migration capacity using a scratch wound healing assay. Immunoprecipitation of methylated DNA combined with real-time PCR was chosen to determine the extent of DNA methylation in HUVEC. Exogenous H2S enhanced angiogenesis in diabetic mice. Exogenous H2S restored endothelial cell migration velocity by upregulating miR-126-3p levels and downregulating high glucose elevation DNMT1 protein levels. Moreover, DNMT1 upregulation in HUVEC increased the methylation level of gene sequences upstream of miR-126-3p, which subsequently inhibited transcription of primary miR-126 and reduced miR-126-3p levels paddy field. Overexpression of CSE in HUVEC rescued miR-126-3p levels by reducing methylation levels and promoting translocation. H2S increases miR-126-3p levels by downregulating methylation levels by reducing high glucose-induced DNMT1 protein levels, thereby ameliorating angiogenesis originally impaired by high glucose.
Diabetes mellitus, hyperglycemia caused by insulin resistance, has become a major global health pandemic. In China, in 2019, he had 110 million diabetics, and that number is expected to rise to 140 million by 2029. Peripheral limb ulceration and amputation due to peripheral vascular disease are common complications of diabetes. The endothelium contains a single layer of cells that lines the lumen of blood vessels and acts as a barrier between the vessel wall and the blood . Endothelial cells perform multiple functions including cell adhesion, angiogenesis, inflammatory response, vascular integrity and regulation of vascular permeability. Dysfunction of vascular endothelial cells by hyperglycemia is a major factor leading to impaired angiogenesis . Endothelial cells are therefore a prime target for promoting angiogenesis. Although ‘therapeutic angiogenesis’ represents a major clinical challenge, pro-angiogenic drugs or gene therapy for ‘therapeutic angiogenesis’ remains the preferred therapy for patients with peripheral vascular disease .
Hydrogen sulfide (H2S) is the third gas transmitter after carbon monoxide and nitrogen oxides. For example, diabetic patients also have low plasma H2S levels, and there is a negative correlation between fasting blood glucose and H2S levels in streptozotocin-induced diabetic rats. Since Kai et al. first reported in 2007 that H2S promotes angiogenesis. Interest in H2S is growing. Over the past decade, H2S has been found to improve cutaneous wound healing in diabetic mice through anti-inflammatory or antioxidant effects, especially by regulating angiogenesis . However, the proangiogenic mechanism of H2S in diabetes needs further investigation. MicroRNAs are a class of noncoding RNAs that are 18-22 nucleotides in length. They also regulate diabetic wound healing through proangiogenesis. B. miR-615-5p and miR-92a. MiR-126-3p is known to regulate endothelial cell angiogenesis. MiR-126 deficiency in mice leads to weak and leaky angiogenesis, abnormal endothelial duct hierarchy, and impaired endothelial cell migration and proliferation . miR-126-3p can repress target genes such as PI3KR2 and SPRED1 and increase angiogenesis. Interestingly, H2S also regulates miRNA transcription, and H2SmicroRNA interactions play an important role in the pathophysiology of cardiovascular disease. Several publications have reported that H2S can reduce ischemia/reperfusioninduced cardiomyocyte apoptosis and combat Parkinson’s disease by modulating microRNAs. However, the relationship between H2S and microRNAs in regulating angiogenesis under high glucose conditions remains unclear.
DNA methylation is an important epigenetic modification that regulates gene expression. DNA methyltransferase 1 (DNMT1) is one of the proteins that maintain DNA methylation. In colon cancer cells, DNMT1 regulates thymosin β 10 (TMSB10) expressions and suppresses tumor growth by maintaining miR-152-3p methylation. Furthermore, DNMT1 functions as an anti-angiogenic protein. DNMT1 protein levels are also associated with diabetes . We found that DNMT1 protein was highly expressed in podocytes or diabetic mice after high glucose treatment. Furthermore, DNMT1 is a potential target to alleviate podocyte injury and diabetic nephropathy. A previous study found that H2S could restore miR-126-3p levels by downregulating her DNMT1 protein levels in human umbilical vein endothelial cells (HUVECs). However, it is unclear how her DNMT1 lowers miR-126-3p during high glucose treatment and whether DNMT1 affects blood flow recovery in mice with her H2S-regulated diabetic hindlimb ischemia . Here, we aimed to investigate how H2S regulates miR-126-3p levels to improve endothelial cell function and promote blood flow in mice with diabetic hindlimb ischemia. Additionally, HUVECs have also been used in in vitro experiments.
Materials and Method
Mice model of type I diabetes
Modeling of Type I Diabetic Mice Male wildtype C57BL/6 mice (22-24 g) were obtained from the SLAC laboratory (Shanghai, China), and the mice were fed with a 12 h light/ dark cycle and water ad libitum Was taken After 2 weeks of acclimatization, 50 mg/kg streptozotocin (Sigma-Aldrich, St. Louis, MO, USA) he administered intraperitoneally for 5 consecutive days to make the mice diabetic. Two weeks later, blood glucose levels were recorded from the animals’ tails after a 4-h fast using a portable glucometer (Johnson, New York, NY, USA). Mice with blood glucose levels above 13.8 mmol/L fell into the group of type I diabetic mice . Mice without STZ treatment were considered as a non-diabetic control group. In this study, we chose only male mice because estrogen directly regulates angiogenesis through effects on endothelial cells and may interfere with the pro-angiogenic effects of H2S.
Model for mice hindlimb ischemia
Briefly, male C57BL/6 mice were anesthetized with 1% sodium pentobarbital and placed on a heating pad (37 °C) to maintain body temperature. The femoral artery was then ligated and an arteriotomy was performed along the medial side of the left hind limb without damaging the vein or nerve. Femoral artery ligation was not performed in sham-operated mice . To confirm whether the model was successful, we immediately monitored blood flow in mice using a MoorLDI2-2 laser Doppler imaging system ‘Moor Instruments, Devon, UK’. Only mice with successful surgery (the ratio of ligated and illuminated contralateral limb measurements was less than 0.1) were used in the following experiments. NaHS (30 and 60 μmol/kg/day; Sigma, St. Louis, MO, USA) was injected intraperitoneally daily for 14 days after ischemia. An equal volume of saline was injected as vehicle .
Exogenous H2S improved angiogenesis in diabetic mice
To determine whether mice with type I diabetes were successfully generated, mice were treated with 50 mg/kg STZ for 5 days followed by a 4-h fast. Diabetic mice with fasting blood glucose levels above 13.8 mmol/L for 4 hours were selected for the following experiments. These mice also exhibited lower body weight compared to non-diabetic controls. Diabetic mice treated with NaHS (30 and 60 μmol/kg/day) for 14 days showed greater blood flow restoration in the hindlimb ischemia model compared to vehicle-treated diabetic mice. Hindlimb ischemia did not affect gastrocnemius weight in diabetic or non-diabetic mice. However, ischemic gastrocnemius muscle weight was decreased in diabetic mice compared with non-diabetic mice. We also monitored capillary density by immunofluorescence, which showed that diabetic mice had lower capillary density in the ischemic gastrocnemius muscle compared with nondiabetic control mice. However, exogenous H2S enhanced diabetes-induced inhibition of capillary density. Furthermore, an in vivo Matrigel plug assay was used to evaluate the pro-angiogenic effect of H2S. Our results showed that hemoglobin content in Matrigel plugs was suppressed in diabetic mice compared with non-diabetic mice, and NaHS treatment restored hemoglobin content in diabetic mice.
Diabetes mellitus is one of the greatest threats to human health. Gangrene and diabetic retinopathy are two of the most common diseases attributed to diabetes, both caused by abnormal angiogenesis. Decreased angiogenic capacity ultimately leads to impaired microcirculation in the lower extremities. As a pro-angiogenic gas mediator, plasma H2S levels are low in diabetes mellitus patients and can enhance angiogenesis by regulating hyperglycemic endothelial cells. The hindlimb ischemia model is a classic in vivo model used to assess the function of angiogenesis. Our results showed that H2S improved blood flow in ischemic diabetic hindlimb mice. Given that capillaries play a key role in angiogenesis, we identified and found that H2S increases capillary density. We also evaluated the proangiogenic effect of her H2S by Matrigel plug His assay. Our results suggest that H2S rescues hyperglycemia-downregulated hemoglobin in Matrigel plugs, which is consistent with previous studies, but little is known about the mechanisms by which this occurs. MicroRNAs are a class of endogenous small noncoding RNAs that can induce mRNA degradation and translational repression by interacting with the 3’UTR, 5’UTR, or CDS of target mRNAs. Because aberrantly expressed microRNAs play an important role in the pathogenesis of microvascular complications, we focused on whether microRNAs are involved in H2Sregulated angiogenesis. By analyzing two sets of microRNA array data, we found that miR-126-3p is regulated by both diabetes and H2S. Zhang et al. reported that miR- 126-3p is a biomarker for diabetes mellitus and can be significantly downregulated in diabetic plasma, confirming that microRNA array data from the GEO database are reliable. . I suggest further. We also confirmed that miR-126-3p levels were downregulated when induced by STZ in diabetic mice, and that exogenous H2S ameliorated miR-126-3p levels in muscle. Indeed, given the plasma microRNA data from patients with type II diabetes, we considered a high-fat model of type II diabetes to be more appropriate. However, it would be better to measure microRNA levels in mouse plasma.
This study focused on the role of H2S in promoting angiogenesis in diabetic mice by downregulating DNA methylation levels. DNMT1 plays an important role in the angiogenic process in diabetic mice. Our results provide new insights into the role of H2S in regulating miR-126-3p under high glucose conditions and suggest new targets for ischemic therapy in diabetes mellitus.
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