Application progress in the role of vitamin D3 in transplantation immunity / 器官移植

Vitamin D3 is a kind of vitamin that plays important roles in maintaining the normal physiological function of the human body, and its metabolites and analogues exhibit strong anti-inflammatory activity. Vitamin D3 could be activated and converted into 1α, 25-dihydroxyvitamin D3, a kind of steroid hormone, in the human body, which participates in the regulation of cellular metabolism by activating vitamin D receptor (a kind of transcription factor), thus exerting immunomodulatory effects. This is essential for maintaining the physiological health of the body. Currently, there is a growing number of studies that suggest important roles for 1α, 25-dihydroxyvitamin D3 in organ transplantation immunomodulation and tolerance. Therefore, we reviewed the overview and physiological effects of 1α, 25-dihydroxyvitamin D3, the immunomodulatory effects of vitamin D3 and the application of vitamin D3 in clinical organ transplantation, and summarized the value of applying vitamin D3 in inducing immune tolerance in transplantation, with the aim of providing a reference for promoting the application of vitamin D3 in transplantation immunity.

Effect of vitamin D drops combined with insulin aspart in the treatment of gestational diabetes mellitus and its influence on serum 1, 25(OH) 2D 3 and RBP4 levels / 中国医师杂志

Objective:To investigate the clinical efficacy of vitamin D drops combined with insulin aspart in the treatment of gestational diabetes mellitus (GDM), and the effect of vitamin D drops on the serum levels of 1, 25 dihydroxyvitamin D 3 [1, 25(OH) 2D 3] and retinol binding protein 4 (RBP4). Methods:A total of 94 GDM patients admitted to the Baoding Second Central Hospital from March 2019 to March 2021 were selected and randomly divided into an observation group and a control group with 47 cases each using a random number table method. The control group received subcutaneous injection of insulin aspartate for treatment, while the observation group received oral vitamin D drops for treatment. After 4 weeks of continuous treatment, the blood glucose control effect and adverse reactions were observed in both groups. The glucose metabolism indicators of the two groups were compared before and after treatment, including fasting blood glucose (FPG), 2-hour postprandial blood glucose (2-hour PG), insulin resistance index (HOMA-IR), and pancreatic islets β Cell Function Index (HOMA-β) and serum levels of 1, 25(OH) 2D 3, RBP4, lipoprotein related phospholipase A2 (Lp-PLA2), and vascular cell adhesion molecule-1 (VCAM-1). All patients were followed up until the end of pregnancy, and Statistical analysis was conducted on the adverse outcomes of two groups of mothers and infants. Results:The time to reach the standard for FPG and 2-hour PG in the observation group, as well as the time for both to reach the standard were significantly shorter than those in the control group (all P<0.05). There was no statistically significant difference in the incidence of dawn phenomenon and hypoglycemia between the observation group and the control group (all P>0.05). After treatment, FPG and 2-hour PG in both groups were significantly reduced compared to those before treatment (all P<0.05); However, after treatment, there was no statistically significant difference between the groups (all P>0.05). Compared with before treatment, HOMA-IR in both groups significantly decreased (all P<0.05), All HOMA- β significantly increased (all P<0.05); And the improvement was more significant in the observation group (all P<0.05). After treatment, the serum levels of 1, 25(OH) 2D 3 in the observation group significantly increased compared to that before treatment ( P<0.05), but there was no significant change in the control group before and after treatment ( P>0.05). After treatment, the levels of serum RBP4, Lp-PLA2, and VCAM-1 in both groups significantly decreased compared to those before treatment (all P<0.05); After treatment, the serum levels of RBP4, Lp-PLA2, and VCAM-1 in the observation group were lower than those in the control group (all P<0.05). The incidence of adverse maternal and neonatal outcomes in the observation group was 14.9%(7/47) and 10.6%(5/47), respectively, which were lower than those in the control group [34.0%(16/47) and 27.7%(13/47)] (all P<0.05). There were 8 cases of hypoglycemia in 94 patients (3 in the observation group and 5 in the control group), and no other adverse events occurred. Conclusions:The combination of vitamin D drops and insulin aspartate in the treatment of GDM can safely, effectively, quickly, and steadily control patients′ blood sugar, improve IR and pancreatic islets β The effect of cell function on reducing the incidence of adverse maternal and fetal outcomes may be related to increasing serum 1, 25(OH) 2D 3 levels and down-regulating the expression levels of serum RBP4, Lp-PLA2, and VCAM-1.

1,25-dihvdroxy vitamin D3 ameliorates neutrophilic asthma by regulating NLRP3/caspase-l/GSDMD signal axis / 中国药理学通报

Aim To investigate the effect of 1,25-dihydroxyvitamin D3 on airway inflammation in a mouse model of neutrophilic asthma and its influence on the NLRP3/caspase-1/GSDMD signal axis.Methods Twenty-four female SPF BALB/c mice were selected and randomly(random number table method)divided into three groups:normal control group(A),model group(B),and intervention treatment group(C),with eight mice in each group.An animal model of mouse neutrophilic asthma was made by the method induced by chicken ovalbumin(OVA)combined with lipopolysaccharide(LPS).Group C was given 0.1% 1,25-(OH)2D3 intraperitoneal injection(4 μg·kg-1)before each OVA challenge.A noninvasive pulmonary function test was performed to assess airway responsiveness,and lung tissues and bronchoalveolar lavage fluid(BALF)were collected.HE staining and AB-PAS staining of lung tissues were performed to observe the pathological changes and airway mucus secretion.Total inflammatory cell count and classification were carried out,and the levels of IL-1β,IL-18,and IL-17 in BALF were detected.Immunohistochemistry staining of Ly-6G was done to confirm the presence of neutrophils in lung.The protein expressions of NLRP3,caspase-1,and GSDMD in lung tissues were determined by Western blot.Results The levels of IL-1β,IL-18,IL-17 in BALF in treatment group was lower than that in model group(P<0.05),and the proportion of neutrophils in BALF was lower than that in model group.Lung histology suggested that airway inflammation in treatment group was less than that in model group.In treatment group,AHR was significantly reduced; NLRP3,caspase-1,GSDMD protein expression levels were significantly lower than those in model group(P<0.05).Conclusions 1,25-dihydroxyvitamin D3 reduces airway inflammation and inflammatory cytokine secretion in mice with neutrophil asthma,and inhibits the expression of NLRP3,caspase-1 and GSDMD in lung.1,25-dihydroxyvitamin D3 may improve neutrophilic asthma by regulating the NLRP3/caspase-1/GSDMD signal axis.

Effects of 1, 25-dihydroxyvitamin D3 on experimental periodontitis and AhR/NF-κB/NLRP3 inflammasome pathway in a mouse model

Abstract Vitamin D has been known to have important regulatory functions in inflammation and immune response and shows inhibitory effects on experimental periodontitis in animal models. However, the potential mechanism has yet to be clarified. Recent studies have highlighted Aryl hydrocarbon receptor (AhR) and its downstream signaling as a crucial regulator of immune homeostasis and inflammatory regulation. Objective: This study aimed to clarify the effect of 1,25-dihydroxyvitamin D3 (VD3) on experimental periodontitis and AhR/nuclear factor-κB (NF-κB)/NLR pyrin domain-containing 3 (NLRP3) inflammasome pathway in the gingival epithelium in a murine model. Methodology: We induced periodontitis in male C57BL/6 wild-type mice by oral inoculation of Porphyromonas gingivalis (P. gingivalis), and subsequently gave intraperitoneal VD3 injection to the mice every other day for 8 weeks. Afterwards, we examined the alveolar bone using scanning electron microscopy (SEM) and detected the gingival epithelial protein using western blot analysis and immunohistochemical staining. Results: SEM images demonstrated that alveolar bone loss was reduced in the periodontitis mouse model after VD3 supplementation. Western blot analyses and immunohistochemical staining of the gingival epithelium showed that the expression of vitamin D receptor, AhR and its downstream cytochrome P450 1A1 were enhanced upon VD3 application. Additionally, VD3 decreased NF-κB p65 phosphorylation, and NLRP3, apoptosis-associated speck-like protein, caspase-1, interleukin-1β (IL-1β) and IL-6 protein expression. Conclusions: These results implicate the alleviation of periodontitis and the alteration of AhR/NF-κB/NLRP3 inflammasome pathway by VD3 in the mouse model. The attenuation of this periodontal disease may correlate with the regulation of AhR/NF-κB/NLRP3 inflammasome pathway by VD3.

Repurposing vitamin D for treatment of human malignancies targeting tumor microenvironment

Tumor cells along with a small proportion of cancer stem cells exist in a stromal microenvironment consisting of vasculature, cancer-associated fibroblasts, immune cells and extracellular components. Recent epidemiological and clinical studies strongly support that vitamin D supplementation is associated with reduced cancer risk and favorable prognosis. Experimental results suggest that vitamin D not only suppresses cancer cells, but also regulates tumor microenvironment to facilitate tumor repression. In this review, we have outlined the current knowledge on epidemiological studies and clinical trials of vitamin D. Notably, we summarized and discussed the anticancer action of vitamin D in cancer cells, cancer stem cells and stroma cells in tumor microenvironment, providing a better understanding of the role of vitamin D in cancer. We presently re-propose vitamin D to be a novel and economical anticancer agent.

1α,25-dihydroxyvitamin D3 inhibits tumor necrosis factor-α induced activation of human umbilical vein endothelial cells via NF-κB signaling pathway / 中华内分泌代谢杂志

Objective To investigate the effect of 1α, 25-dihydroxyvitamin D3 [1α,25-(OH)2 D3] on tumor necrosis factor-α ( TNF-α) induced activation of human umbilical vein endothelial cells ( HUVECs ) . The mechanism involved in this process was also studied. Methods HUVECs were cultured and treated with TNF-α( 40 ng/ml), 1α,25-(OH)2D3(10-8 mol/L), and SN50 as indicated. Vascular cell adhesion molecule (VCAM) and E-selectin were used as markers of endothelial activation, which were detected by Western blotting and realtime PCR (RT-PCR). NF-κB signaling pathway was investigated to study the mechanism. Western blotting, RT-PCR, immunofluorescence assay, and chromatin immunoprecipitation ( ChIP ) method were used to evaluate the effects of 1α,25-( OH) 2 D3 on its early activation, nuclear transport, and binding to VCAM and E-selectin promoters. Results (1) Western blotting and RT-PCR showed that TNF-α could significantly up-regulate the expression of VCAM and E-selectin in HUVECs, which can be inhibited by specific NF-κB blocker SN50. 1α,25-( OH) 2 D3 down-regulated the expression of VCAM and E-selectin induced by TNF-α. ( 2 ) Western blotting showed that TNF-α induces I-κBαphosphorylation, thereby activating NF-κB p65 subunit. Immunofluorescence showed that 1α, 25-( OH ) 2 D3 significantly inhibited the nuclear translocation of NF-κB p65 subunit. ChIP analysis revealed that 1α,25-( OH) 2 D3 inhibited the binding of NF-κB p65 to VCAM and E-selectin promoters and thus affected gene expression. Conclusions TNF-αenhanced the expression of E-selectin and VCAM in HUVECs via NF-κB signaling pathway. 1α,25-( OH) 2 D3 may inhibit NF-κB early activation, nuclear transport and the binding of NF-κB p65 to VCAM and E-selectin promoters, thereby inhibiting TNF-α-induced endothelial cell activation.

1,25-Dihydroxyvitamin D3 protects obese rats from metabolic syndrome promoting regulatory T cell-mediated resolution of inflammation

Vitamin D has been found to produce therapeutic effects on obesity-associated insulin resistance and dyslipidemia through its potent anti-inflammatory activity, but the precise immunomodulatory mechanism remains poorly understood. In the present study we found that 1,25-dihydroxyvitamin D [1,25(OH)D], the biologically active form of vitamin D, significantly attenuated monosodium glutamate (MSG)-induced obesity and insulin resistance as indicated by body weight reduction, oral glucose tolerance improvement, and a glucose infusion rate increase as detected with hyperinsulinemic-euglycemic clamp. Moreover, 1,25(OH)D not only restored pancreatic islet functions but also improved lipid metabolism in insulin-targeted tissues. The protective effects of 1,25(OH)D on glycolipid metabolism were attributed to its ability to inhibit an obesity-activated inflammatory response in insulin secretory and targeted tissues, as indicated by reduced infiltration of macrophages in pancreas islets and adipose tissue while enhancing the expression of in liver tissue, which was accompanied by increased infiltration of Treg cells in immune organs such as spleen and lymph node as well as in insulin-targeted tissues such as liver, adipose, and muscle. Together, our findings suggest that 1,25(OH)D serves as a beneficial immunomodulator for the prevention and treatment of obesity or metabolic syndrome through its anti-inflammatory effects.

Effect of 1,25-dihydroxyvitamin D3 on renal expression of TGF-1/Smads in diabetic nephropathy rats / 中国基层医药

Objective To explore the effects of 1,25-dihydroxyvitamin D3 on renal expression of TGFβ1,Smad3 and Smad7.Methods 40 Sprague-Dawley mts were randomized into 4 groups:normal control rats (group A),diabetic nephropathy group (group B),small dose 1,25-dihydroxyvitamin D treatment group (0.5 μg · kg-1 · d-1,group C) and large dose 1,25-dihydroxyvitamin D treatment group (1 μg · kg-1 · d-1,group D),each group had 10 rats.After 12 weeks,the renal function,blood glucose,glycosylated hemoglobin and the urine trace albumin content of each rats were tested.Results The biochemical indicators in group B were higher than those in group A,(t =-16.566,P ＜0.05;t =-16.949,P ＜0.05;t =-11.844,P ＜0.05;t =-19.778,P ＜0.05;t =-14.013,P ＜ 0.05).Compared with group B,the biochemical indicators and the expression of TGFβ1,Smad3 mRNA reduced in group C and group D,and the expression of Smad7 mRNA increased (F =37.892,P ＜ 0.05;F =70.068,P ＜ 0.05;F =21.95,P ＜0.05;F =77.619,P ＜0.05;F =37.670,P ＜0.05;F =1062.562,P ＜0.05;F =2463.789,P ＜0.05;F =81.745,P ＜ 0.05).There were no significant differences between group C and group D (t =0.538,P＞0.05;t =1.737,P＞0.05;t =0.671,P＞0.05;t =1.763,P ＞0.05;t =0.997,P ＞0.05;t =1.653,P ＞0.05;t=1.543,P＞0.05;t =-1.313,P ＞0.05).Conclusion 1,25-dihydroxyvitamnin D3 has protective effect on diabetic nephropathy rats model,the mechanism may be associated with inhibiting the expression of TGF β1 and Smad3,increasing the expression of Smad7.

1α,25-Dihydroxyvitamin D3 Inhibits Activation of Hepatic Stellate Cells by Up-regulating MiR-146a Expression / 胃肠病学

Background:1 α,25-dihydroxyvitamin D3 [1,25 (OH) 2 D3],the active form of vitamin D,is reported in some studies having antifibrotic potential in liver fibrosis,however,its mechanism is not fully clarified.MicroRNAs (miRNAs) have recently been shown could regulate the proliferation and activation of hepatic stellate cells (HSCs),and are involved in the promotion or inhibition of liver fibrosis.Aims:To explore whether the inhibiting effect of 1,25 (OH) 2 D3 on activation of HSCs is by regulating miRNAs expression.Methods:Literature review and qPCR method were used to screen out the differentially expressed miRNAs between transforming growth factor-β1 (TGF-β1)-stimulated (activated) HSCs and the inactivated HSCs.Then the HSCs were co-cultured with TGF-β1 and the mimic of differentially expressed miRNA,the negative control mimic,1,25(OH)2D3 and DMSO,respectively,and the cell viability and apoptosis were determined by CCK-8 assay and flow cytometry.Results:Expression of miR-146a was down-regulated in activated HSCs (P ＜ 0.05).Compared with HSCs in DMSO group,the expression of miR-146a was significantly up-regulated in HSCs treated with 1,25 (OH) 2 D3;meanwhile,the cell viability was decreased and the apoptosis was increased (P all ＜ 0.05).In HSCs transfected with miR-146a mimic,the expression of miR-146a was up-regulated,the cell viability was decreased,and the apoptosis was increased similarly with HSCs in 1,25 (OH)2D3 group (P all ＜ 0.05).Conclusions:Regulation of miR-146a expression might be one of the important mechanisms of 1,25 (OH) 2 D3 in inhibiting TGF-β1-stimulated HSC activation and inducing apoptosis in HSCs.

Associations between serum 25-hydroxyvitamin D3 level and skeletal muscle mass and lower limb muscle strength in Japanese middle-aged subjects

OBJECTIVES: One of the important risk factors of falling is decreased muscle mass and muscle strength. Recently, there has been an increasing concern on the role of vitamin D in muscle strength and physical activity. Aim of our study is to examine the relationships between vitamin D status and muscle mass and muscle strength in middle-aged healthy adults. METHODS: Subjects were 40 healthy volunteers aged 42.0 ± 10.6 years old. Evaluation was made for serum vitamin D₃ metabolites including 25-hydroxyvitamin D₃ [25(OH)D₃] and 24,25-dihydroxyvitamin D₃ [24,25(OH)₂D₃] concentrations, lower limb muscle strength, and dietary intake by food frequency questionnaire. Body composition was measured by dual-energy X-ray absorptiometry (DXA), and appendicular skeletal mass index (ASMI) was calculated as skeletal muscle mass/squared height. RESULTS: 70% of the subjects had vitamin D insufficiency/deficiency (serum total 25(OH)D < 20 ng/mL), and female subjects had significantly lower serum total 25(OH)D level compared with males. Vitamin D insufficiency/deficiency group had significantly higher body fat, lower SMI and muscle strength, probably reflecting higher percentage of female subjects. Serum vitamin D₃ metabolites levels were significantly correlated with whole and site-specific ASMI, and lower limb muscle strength, except for the correlation between serum 24,25(OH)₂D₃ concentration and lower limb muscle strength. In addition, serum 25(OH)D₃ level was a positive significant predictor for both ASMI and lower limb muscle strength, while serum 24,25(OH)₂D₃ level was not their significant predictor. CONCLUSIONS: Serum 25(OH)D₃ level was significantly correlated with both skeletal muscle mass and lower limb muscle strength.

mTOR signaling pathway for regulating inhibition of 1,25 dihydroxyvitamin D3 on Hep-2 cells proliferation in laryngeal carcinoma / 国际检验医学杂志

Objective To explore the inhibition role of 1 ,25 dihydroxyvitamin D3 on laryngeal cancer Hep‐2 cell proliferation and its influence on mTOR signal pathway .Methods Hep‐2 cells were treated with different concentrations of 1 ,25 dihydroxyvitamin D3 (10-8 ,10-7 ,10-6mol/L) for 24 ,48 ,72 h respectively .The proliferation situation of Hep‐2 cells was detected by the MTT meth‐od and the inhibition rate was calculated .The effect of 1 ,25 dihydroxyvitamin D3 on Hep‐2 cell cycle distribution was analyzed by flow cytometry .The influence of 1 ,25 dihydroxyvitamin D3 on mTOR signaling pathway was detected by Western blot .Results Different concentrations of 1 ,25 dihydroxyvitamin D3 could inhibit the proliferation of Hep‐2 cells ,changed the cell cycle distribu‐tion and increased the proportion of Hep‐2 cells in G0/G1 phase .The expressions of TSC1 and TSC2 protein after 1 ,25 dihydroxyvi‐tamin D3 intervention were increased compared with the control group (P<0 .01) ,while the Rheb protein expression was signifi‐cantly decreased(P<0 .01):mTOR protein and phosphorylation level were significantly decreased compared with the control group (P<0 .01) ,the decrease of mTOR protein phosphorylation was especially obvious (P<0 .01);4EBP‐1 protein expression was in‐creased compared with the control group (P<0 .01) .Conclusion 1 ,25‐dihydroxyvitamin D3 alters the Hep‐2 cell cycle distribution , affects the protein expression of mTOR signaling pathway ,thus inhibits the cell proliferation .

The expression and clinical significance of serum 1, 25-dihydroxyvitamin D3 in different stages of diabetic nephropathy patients / 中国综合临床

Objective To study the expression and clinical significance of serum 1,25-dihydroxyvitamin D3 in different stages of diabetic nephropathy patients.Methods Ninety-eight cases of diabetics were selected as our subjects in observation group,who were hospitalized in Tieying Hospital of Fengtai District of Beijing from January 2010 to December 2014.They were divided into diabetics group(36 cases,UAER ＜30 mg/24 h),microalbuminuria group(32 cases,UAER was 30-300 mg/24 h),massive proteinuria group (30cases,UAER＞300 mg/24 h).One hundred healthy persons were selected as a normal control group over the same period.The changes of fasting blood glucose,course of disease,blood lipid,serum creatinine and serum 1,25-dihydroxyvitamin D3 in all groups were recorded.Results Fasting blood glucose,serum creatinine and serum 1,25-dihydroxyvitamin D3 were (8.97±2.3) mmol/L,(76.2±19.5) μmol/L,(28.9±13.6) μg/L in observation group,and (4.7±0.4) mmol/L,(63.1±12.0) μmol/L,(70.1±21.3) μg/L in control group respectively,the difference between the two group was statistically significant (P =0.031,0.046,0.028).Serum 1,25-dihydroxyvitamin D3 was (52.68±20.91) μg/L in patients of diabetics group,(31.40±15.23) μg/L in microabuminuria group,(15.76±7.81) μg/L in massive proteinuria group,the difference among the three group was statistically significant (P =0.036).Serum 1,25-dihydroxyvitamin D3 of microabuminuria group and massive proteinuria group were lower than of diabetics group,of massive proteinuria group was lower than of microabuminuria group(P＜0.05).Serum 1,25-dihydroxyvitamin D3 was negatively correlated with the cause of disease(r=-0.301),fasting blood glucose (r =-0.281) and serum creatinine (r =-0.536) in patients with type 2 diabetes,the difference was statistically significant (P ＜ 0.05).Conclusion Serum 1,25-dihydroxyvitamin D3 in patients with diabetic nephropathy decrease in different degree,which reflects the severity of renal damage.The results indicate that reduction of serum 1,25-dihydroxyvitamin D3 may be involved in the occurrence and development of diabetic nephropathy

The Association of Low-energy Spine Fractures and Vitamin D Inadequacy: A Case-control Study

STUDY DESIGN: Retrospective study. OBJECTIVES: To compare serum vitamin D levels in elderly patients with or without osteoporotic spinal compression fractures (OSCFs) and to identify relationships between the serum vitamin D level and other variables, such as age, bone mineral density (BMD), and bone turnover markers (osteocalcin and C-telopeptide). SUMMARY OF LITERATURE REVIEW: Vitamin D plays a key role in calcium metabolism in the bone tissue. Vitamin D deficiency can lead to decreased BMD and an increased risk of falls and of osteoporotic fractures. MATERIALS AND METHODS: We retrospectively reviewed the medical records of 95 elderly patients (≥60 years) with OSCFs (fracture group) and 118 subjects who had been diagnosed with osteoporosis without OSCFs (control group). Serum vitamin D levels were contrasted between the two groups taking into account other factors such as patient age, sex, and seasonal variations. For all the patients, we also evaluated the correlation between the vitamin D level and the patient age, BMD, and bone turnover markers. RESULTS: The mean of the serum 25(OH) vitamin D3 levels was significantly lower in the fracture group than in the control group. There were significant differences in the 25(OH) vitamin D3 levels in autumn. In all patients, the mean serum 25(OH) vitamin D3 levels were the highest in autumn and the lowest in spring. Furthermore, the mean serum 25(OH) vitamin D3 levels were significantly correlated with patient age and BMD. CONCLUSIONS: A low serum vitamin D level might be a risk factor of OSCFs in elderly patients.

Treatment with 1,25(OH)2D3 induced HDAC2 expression and reduced NF-κB p65 expression in a rat model of OVA-induced asthma

Recent evidence indicates that a deficiency of 1,25-dihydroxyvitamin D3 (1,25[OH]2D3) may influence asthma pathogenesis; however, its roles in regulating specific molecular transcription mechanisms remain unclear. We aimed to investigate the effect of 1,25(OH)2D3 on the expression and enzyme activity of histone deacetylase 2 (HDAC2) and its synergistic effects with dexamethasone (Dx) in the inhibition of inflammatory cytokine secretion in a rat asthma model. Healthy Wistar rats were randomly divided into 6 groups: control, asthma, 1,25(OH)2D3 pretreatment, 1,25(OH)2D3 treatment, Dx treatment, and Dx and 1,25(OH)2D3 treatment. Pulmonary inflammation was induced by ovalbumin (OVA) sensitization and challenge (OVA/OVA). Inflammatory cells and cytokines in the bronchoalveolar lavage (BAL) fluid and histological changes in lung tissue were examined. Nuclear factor kappa B (NF-κB) p65 and HDAC2 expression levels were assessed with Western blot analyses and quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR). Enzyme activity measurements and immunohistochemical detection of HDAC2 were also performed. Our data demonstrated that 1,25(OH)2D3 reduced the airway inflammatory response and the level of inflammatory cytokines in BAL. Although NF-κB p65 expression was attenuated in the pretreatment and treatment groups, the expression and enzyme activity of HDAC2 were increased. In addition, 1,25(OH)2D3 and Dx had synergistic effects on the suppression of total cell infusion, cytokine release, and NF-κB p65 expression, and they also increased HDAC2 expression and activity in OVA/OVA rats. Collectively, our results indicated that 1,25(OH)2D3 might be useful as a novel HDAC2 activator in the treatment of asthma.

The mechanism played by 1,25-dihydroxyvitamin D3 in treating renal fibrosis in diabetic nephropathy / 中华内分泌代谢杂志

Objective To observe the effects of 1,25-dihydroxyvitamin D3 on the expressions of transforming growth factor-β1(TGF-β1), fibronectin(FN),and vascular endothelial growth factor(VEGF) in rats with diabetic nephropathy(DN), and to elucidate the protective mechanism played by 1,25-dihydroxyvitamin D3. Methods DN models were estabolished by injecting streptozotoein ( STZ ) into male SD rats, which were divided into TGF-β1 overexpression group, TGF-β1 overexpression plus vitamin D3 group, TGF-β1 low-expression group, TGF-β1 low-expression plus vitamin D3 group, TGF-β1 normal-expression group, and TGF-β1 normal-expression plus vitamin D3 group. After 1,25-dihydroxyvitamin D3 treatment for 37 days, renal function and blood biochemical parameters were evaluated. The morphology and fibrosis of kidney tissues were observed. The expressions of TGF-β1, FN, and VEGF in kidney cortex were measured by immunohistochemistry, realtime PCR, and Western blotting. Results The levels of cholesterol, triglyceride, creatinine,plasma glucose, HbA1C , and 24 h urinary protein were lower in vitamin D3treated groups than those in corresponding control groups(P<0. 05). The degree of renal fibrosis was raised with the increased level of TGF-β1. Vitamin D3 treatment decreased the fibrosis in diabetic kidney. There were significant differences in the mRNA and protein expressions of TGF-β1 in three control groups(P<0. 05). With the increased levels of TGF-β1, the expressions of FN and VEGF were increased. The expressions of TGF-β1, FN, and VEGF were lowered by vitamin D3compared with the corresponding control groups(P<0. 05). Conclusion 1,25-dihydroxy-vitamin D3 may protect the renal tissure in diabetic rats via inhibiting the expressions of TGF-β1, FN, and VEGF in the kidney.

Effect of 1, 25-dihydroxyvitamin D3 on mast cells tryptase in asthmatic guinea pigs

Objective:To explore the effect of 1,25-dihydroxyvitamin D3 on the mast cell tryptase (MCT) in asthmatic guinea pigs. Methods:A total of 60 male or female healthy guinea pigs were randomly divided into control group (group A), asthmatic group(group B), and 1,25-dihydroxyvitamin D3 group(group C), with 20 cases in each group. To establish asthmatic guinea pig models, 1ml peanut oil was filled into stomach in the morning in group A and group B , and 1ml peanut oil with 1,25-dihydroxyvitamin D3 was filled into stomach in group C. Airway resistance (Re) of asthmatic guinea pigs was detected, and the bronchoalveolar lavage fluid (BALF) cells were counted. Lung tissue with HE and MCT immunohistochemical staining were used to observe the pathological changes in lung tissue and the distribution of MCT. Results:After injection of different concentration of acetylcholine chloride, the Re in group B and group C were increased significantly compared with group A (P<0.05);compared with group B, the Re in group C were decreased significantly (t=-5.385, -5.761, -6.184,-13.574, P<0.05);the total number of BALF cells and eosinophils were increased significantly in group B and C (t=19.618, 9.598, 10.854, 5.388, P<0.05);compared with group B, the total number of BALF cells and eosinophils in group C was decreased significantly (t=-5.555,-5.392, P<0.05);the number of tryptase positive cells in group B was increased significantly than that in group A (t=21.312, P<0.05), and in addition to the alveolar septum and submucosa, the cells were also distributed around blood vessels and outside the cells; the number of tryptase positive cells in group C was decreased significantly compared with group B, and the difference was statistically significant (t=5.043, P<0.05). Conclusions:After the asthmatic guinea pigs are treated with 1,25-dihydroxyvitamin D3, their BALF, Re, infiltration degree of inflammatory cells in the trachea and lung tissue and airway inflammatory reaction are reduced significantly. 1,25-dihydroxyvitamin D3has a certain inhibiting effect on the activation of mast cells and the release of MCT granules.

Effect of 1,25-dihydroxyvitamin D3 on mast cells tryptase in asthmatic guinea pigs

Objective: To explore the effect of 1,25-dihydroxyvitamin D3 on the mast cell tryptase (MCT) in asthmatic guinea pigs. Methods: A total of 60 male or female healthy guinea pigs were randomly divided into control group (group A), asthmatic group (group B), and 1,25-dihydroxyvitamin D3 group (group C), with 20 cases in each group. To establish asthmatic guinea pig models, 1mL peanut oil was filled into stomach in the morning in group A and group B, and 1mL peanut oil with 1,25-dihydroxyvitamin D3 was filled into stomach in group C. Airway resistance (Re) of asthmatic guinea pigs was detected, and the bronchoalveolar lavage fluid (BALF) cells were counted. Lung tissue with HE and MCT immunohistochemical staining were used to observe the pathological changes in lung tissue and the distribution of MCT. Results: After injection of different concentration of acetylcholine chloride, the Re in group B and group C were increased significantly compared with group A (P<0.05); compared with group B, the Re in group C were decreased significantly (t=-5.385,-5.761,-6.184,-13.574, P<0.05); the total number of BALF cells and eosinophils were increased significantly in group B and C (t=19.618, 9.598, 10.854, 5.388, P<0.05); compared with group B, the total number of BALF cells and eosinophils in group C was decreased significantly (t=-5.555,-5.392, P<0.05); the number of tryptase positive cells in group B was increased significantly than that in group A (t=21.312, P<0.05), and in addition to the alveolar septum and submucosa, the cells were also distributed around blood vessels and outside the cells; the number of tryptase positive cells in group C was decreased significantly compared with group B, and the difference was statistically significant (t=5.043, P<0.05). Conclusions: After the asthmatic guinea pigs are treated with 1,25-dihydroxyvitamin D3, their BALF, Re, infiltration degree of inflammatory cells in the trachea and lung tissue and airway inflammatory reaction are reduced significantly. 1,25-dihydroxyvitamin D3 has a certain inhibiting effect on the activation of mast cells and the release of MCT granules.

Effect of 1,25-dihydroxyvitamin D3 on mast cells tryptase in asthmatic guinea pigs

OBJECTIVE@#To explore the effect of 1,25-dihydroxyvitamin D3 on the mast cell tryptase (MCT) in asthmatic guinea pigs.@*METHODS@#A total of 60 male or female healthy guinea pigs were randomly divided into control group (group A), asthmatic group (group B), and 1,25-dihydroxyvitamin D3 group (group C), with 20 cases in each group. To establish asthmatic guinea pig models, 1 mL peanut oil was filled into stomach in the morning in group A and group B, and 1 mL peanut oil with 1,25-dihydroxyvitamin D3 was filled into stomach in group C. Airway resistance (Re) of asthmatic guinea pigs was detected, and the bronchoalveolar lavage fluid (BALF) cells were counted. Lung tissue with HE and MCT immunohistochemical staining were used to observe the pathological changes in lung tissue and the distribution of MCT.@*RESULTS@#After injection of different concentration of acetylcholine chloride, the Re in group B and group C were increased significantly compared with group A (P < 0.05); compared with group B, the Re in group C were decreased significantly (t = -5.385, -5.761, -6.184, -13.574, P < 0.05); the total number of BALF cells and eosinophils were increased significantly in group B and C (t = 19.618, 9.598, 10.854, 5.388, P < 0.05); compared with group B, the total number of BALF cells and eosinophils in group C was decreased significantly (t = -5.555, -5.392, P < 0.05); the number of tryptase positive cells in group B was increased significantly than that in group A (t = 21.312, P < 0.05), and in addition to the alveolar septum and submucosa, the cells were also distributed around blood vessels and outside the cells; the number of tryptase positive cells in group C was decreased significantly compared with group B, and the difference was statistically significant (t = 5.043, P < 0.05).@*CONCLUSIONS@#After the asthmatic guinea pigs are treated with 1,25-dihydroxyvitamin D3, their BALF, Re, infiltration degree of inflammatory cells in the trachea and lung tissue and airway inflammatory reaction are reduced significantly. 1,25-dihydroxyvitamin D3 has a certain inhibiting effect on the activation of mast cells and the release of MCT granules.

Improved induction culture system for Raw264.7 cells to differentiate into osteoclasts / 吉林大学学报(医学版)

Objective To establish a high-performance induction culture system for Raw264.7 cells to differentiate into osteoclasts(OC)invitro by improving the cell culture program.Methods The Raw264.7 cells were cultured withα-MEM medium containing 50 μg · L-1 M-CSF, 100 μg · L-1 RANKL, and 1 × 10-8 mol · L-1 1α,25-(OH)2 D3 in 5% CO2 for 12 d at 37℃. The cells were digested transiently every time before the medium was changed after every three days. The morphologic changes of the Raw264.7 cells were observed by inverted microscope.The maturation and phagotrophic function of OC were identified by HE,TRAP,FITC-phalloidin staining and immunofluorescence.Results The cells remained to grow in single layers all the time in most areas of the well during the whole induction by the improved culture program. The observation results of inverted microscope and HE staining showed that the growth area of the polykaryotic OC reached to 70% of the well on day 1 2. FITC-phalloidin staining showed that in the maturation of the OC, the cluster-shaped podosomes in the pseudopodia gradually transformed into rings,which finally fused to form a large belt surrounding the periphery of the cytoplasm. The calcitionin receptor (CTR) expressed by OC was markedly enhanced compared with the precursor cells by immunofluroescence staining,and a large number of red granules appeared in the cytoplasm of OC with TRAP staining on day 1 2. These results comfirmed that the obtained OC were maturated and owned phagotrophic function. Conclusion A high-performance induction culture system for Raw264. 7 cells to differentiate into OC in vitro induced by combination of 50μg · L-1 M-CSF, 100 μg · L-1 RANKL,and 1 × 10-8 mol·L-1 1α,25-(OH)2 D3 is established by improving the cell culture program.

Effect of 1,25-dihydroxyvitamin D3 on interleukin 17 expression in mice with asthma / 中华实用儿科临床杂志

Objective To explore the effects of interleukin 17 (IL-17) and 1,25-dihydroxyvitamin D3 [1,25-(OH)2VD3] on the pathogenesis of asthma.Methods Thirty-two Kunming mice were randomly divided into 4 groups:the control group (group A),the asthma group (group B),the 1,25-(OH)2VD3-treated group (group C) and the Dexamethasone-treated group (group D).The allergic mouse models were established by using ovalbumin (OVA).The behavioral changes of mice were observed and the number of leukocytes in the bronchoalveolar lavage fluid (BALF) were counted.HE staining was used to measure the histological changes of lung tissue.The levels of IL-17 and IgE in the BALF were detected by enzyme-linked immunosorbent assay (ELISA).Results The asthmatic symptoms were more severe in group A than those in the other groups,while they were relieved significantly in group C and group D when treated with 1,25-(OH)2VD3 or Dexamethasone.The number of leukocyte in groups B,C and D [(1.97 ± 0.23) × 108/L,(1.02 ±0.17) × 108/L,(0.95 ±0.14) × 108/L]were higher than those in group A[(0.56 ±0.16) × 108/L] (F =85.58,P ＜ 0.01),and lower in group C and group D than those in group B (all P ＜ 0.01),but there were no significant differences between group C and group D(P ＞0.05).The levels of IgE and IL-17 in group B were significantly higher than those in group A(all P ＜ 0.01).The levels of IgE and IL-17 in group B were significantly lower than those in group C and group D,but higher than those in group A(all P ＜0.01).No significant results were observed between group C and group D (all P ＞ 0.05).Conclusions IL-17 involves in the inflammation process of asthma,which promotes the respiratory inflammation.1,25-(OH)2VD3 may suppress IL-17 expression to relieve the respiratory inflammation in acute asthmatic mice,the effect of which is similar to that of Dexamethasone on asthma.