IL-17A inhibits the degradation of RANKL in osteoblasts by inhibiting BCL2-Beclin1-autophagy signaling.

The inflammatory cytokine IL-17A is known to have the capacity to promote osteoclastogenesis, thereby enhancing bone loss. Moreover, IL-17A can promote the expression of RANKL in osteoblasts, contributing to its pro-osteoclastogenic effect. IL-17A is an autophagy regulator, which is also responsible for its regulation on RANKL expression. However, the specific role of autophagy in IL-17A-regulated RANKL expression and the underlying mechanism of IL-17A-regulated osteoblast autophagy remain unclear. IL-17A is known to inhibit autophagy by preventing BCL2 degradation. This study aimed to explore the significance of BCL2-dependent autophagy in IL-17A-regulated RANKL expression. Our results showed that IL-17A at 50 ng/mL could inhibit autophagic activity and promote RANKL protein expression in MC3T3-E1 osteoblast line. Moreover, the corresponding concentration of IL-17A could enhance BCL2 protein expression and the protein interaction between BCL2 and Beclin1 in MC3T3-E1 cells. However, the protein expression of RANKL and BCL2 promoted by 50 ng/mL of IL-17A was blocked by autophagy activation with Beclin1 pharmacological upregulation. Furthermore, RANKL protein expression promoted by 50 ng/mL of IL-17A was also reversed by autophagy activation with BCL2 knockdown. Importantly, the supernatant from osteoblasts treated with 50 ng/mL of IL-17A made osteoclast precursors (OCPs) form larger osteoclasts, which was reversed by BCL2 knockdown in osteoblasts. In conclusion, high levels of IL-17A prevent the degradation of RANKL by inhibiting BCL2-Beclin1-autophagy activation signal transduction in osteoblasts, thereby indirectly promoting osteoclastogenesis.

[Serum free fatty acid level in children with primary hypertension]. / å„¿ç«¥åŽŸå‘æ€§é«˜è¡€åŽ‹è¡€æ¸ æ¸¸ç¦»è„‚è‚ªé ¸çš„æ°´å¹³åŠæ„ä¹‰.

OBJECTIVES: To examine the serum level of free fatty acid (FFA) in children with primary hypertension and its value in the pathogenesis, prevention, and treatment of primary hypertension in children. METHODS: In this retrospective study, 34 children with primary hypertension who were treated for the first time in Children's Hospital Affiliated to Capital Institute of Pediatrics from January to June, 2021, were enrolled as the hypertension group, and 32 children with normal blood pressure who underwent physical examination during the same period were enrolled as the control group. The two groups were compared in terms of the levels of fasting serum FFA, fasting serum triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and non-high-density lipoprotein cholesterol (non-HDL-C). The multivariate logistic regression model was used to analyze the influence of FFA on the development of primary hypertension. RESULTS: Compared with the control group, the hypertension group had significantly higher body mass index (BMI), systolic blood pressure, and diastolic blood pressure (P<0.05), as well as significantly higher serum levels of FFA, TG, LDL-C, and non-HDL-C and a significantly lower serum level of HDL-C (P<0.05). Compared with the control group, the hypertension group had significantly higher rates of elevated serum FFA (>0.45 mmol/L for girls and >0.60 mmol/L for boys) (P<0.05) and abnormal blood lipid levels (abnormality in at least one index among serum TG, TC, LDL-C, HDL-C, and non-HDL-C) (P<0.05). A multivariate logistic regression equation was established based on age, sex, BMI, elevated serum FFA, and abnormal blood lipid levels, and the results showed that elevated serum FFA was an independent risk factor for primary hypertension in children (OR=17.560, 95%CI: 1.964-157.003, P<0.05). CONCLUSIONS: There is a significant increase in serum FFA level in children with primary hypertension, and the increase in serum FFA can increase the risk of primary hypertension in children.

Increasing angiotensin-converting enzyme 1 regulated by histone 3 lysine 27 hyperacetylation in high-fat diet-induced hypertensive rat kidney.

BACKGROUND: Obesity is a key risk factor of hypertension. Angiotensin-converting enzyme 1 (ACE1) is a key enzyme involved in the renin-angiotensin-aldosterone system (RAAS), which contributes to obesity-related hypertension (OrHTN). Emerging evidence has shown that histone acetylation is also involved in OrHTN. As kidney is an effector organ that activates the RAAS by secreting renin after hypertension occurs, this study aimed to explore the regulatory role of histone acetylation on renal RAAS expression. METHODS: Nineteen male Wistar rats were randomly divided into a control group ( n  = 9, fed normal chow) and a high-fat diet (HFD) group ( n  = 10, fed HFD for 16 weeks). The renal transcriptome and histone acetylation spectrum was analyzed by RNA sequencing and tandem mass spectrometry and was further confirmed by RT-qPCR, western blot, and immunohistochemistry. Then, chromatin immunoprecipitation (ChIP)-qPCR analysis was performed for the detection of DNA-protein interaction. RESULTS: After 16-week HFD, the rats became obese with increased plasma triglyceride and high blood pressure. Increased ACE1 and histone 3 lysine 27 acetylation (H3K27ac) expression levels were found in OrHTN rat kidneys. The following ChIP-qPCR analysis illustrated that the upregulation of ACE1 transcription was mediated by increased H3K27ac. CONCLUSION: H3K27ac could be an important histone acetylation site that activates renal ACE1 in HFD-induced hypertensive rats.

Nbr1-regulated autophagy in Lactoferrin-induced osteoblastic differentiation.

The molecular mechanism of autophagy in Lactoferrin (LF) induced osteoblast differentiation is not fully demonstrated. In this study, alkaline phosphatase (ALP) activity, alizarin red S staining and ELISA were used to study N-terminal propeptide of type I procollagen (PINP) expression. mRFP-GFP-LC3 adenoviruses, mono-dansylcadaverine (MDC) staining, scanning electron microscopy, and western blot analysis was employed to probe the LF induced autophagy. The interaction between autophagy receptor Neighbor of Brca1 gene (Nbr1) and pp38 was studied. 3-methyladenine (3-MA) and chloroquine (CQ) could inhibit the activity of ALP, PINP and the autophagy in LF group. LF treatment could up-regulate and down-regulate the expressions of pp38 and Nbr1with a dose-dependent manner, respectively. LF could inhibit the recognition of pp38 and Nbr1. In addition, LF can prompt Nbr1-medicated autophagy and prevent pp38 degradation by autophagy. LF can induce Nbr1-mediated autophagy and inhibit pp38 entering into autophagy flux in the physiological process of osteoblast differentiation.Abbreviations: CQ:chloroquine；LF: Lactoferrin; 3-MA: 3-methyladenine; ALP: Alkaline phosphatase; ANOVA: Analysis of variance; CCK-8: Cell Counting Kit-8; LC3: Microtubule-associated protein light chain3; MDC: Monodansylcadaverine; Nbr1: neighbor of Brca1 gene; PINP: N-terminal propeptide of type I procollagen; PVDF: Polychlorotrifluoroethylene; pp38: phosphorylation p38; RAPA: Rapamycin; SDS: sodium dodecyl sulfate.

MicroRNA-143 promotes apoptosis of osteosarcoma cells by caspase-3 activation via targeting Bcl-2.

Osteosarcoma is the most common malignant bone tumor. In recent years, although a lot of research in the mechanism of osteosarcoma development and metastasis had been done, the molecular mechanisms are still elusive. MicroRNAs (miRs), as small noncoding RNA sequences, are dysregulated in various diseases, including cancer, negatively modulating the target genes expression by posttranscriptional repression. MicroRNA-143 (miR-143) has been reported to be reduced in cancers, including pituitary, colorectal, prostate cancer and cervical. We were aimed to detect the effects of miR-143 on osteosarcoma cell invasion and migration as well as to indicate the potential molecular mechanisms by which miR-143 regulated osteosarcoma. After miR-143 transfection, the cancer cells migration and invasion were examined. And Western blot, RT-PCR, flow cytometry and immunochemistry assays were performed to analyze the role of miR-143 in osteosarcoma progression. The results suggested that miR-143 expressed lessly in osteosarcoma cell lines and could suppress cell migration and invasion in U2-OS and MG-63 cells. To our knowledge, it was the first time to target Bcl-2 directly to explore the underlying mechanism by which miR-143 performed its role to induce apoptosis in tumor cells, thus improving osteosarcoma progression. The present study indicated that miR-143 could inhibit Bcl-2 expression, causing Caspas3 activation, thus inducing apoptosis in osteosarcoma cells. MiR-143 may therefore sreve as a potential biomarker for osteosarcoma, and the regulation of its expression might be a novel therapeutic strategy for osteosarcoma treatment.

The relationship between oxidative stress and the levels of serum circulating adhesion molecules in patients with hyperglycemia crises.

OBJECTIVE: To investigate the relationship between oxidative stress and the levels of serum circulating adhesion molecules in patients with hyperglycemia crises. METHODS: A total of 73 patients with diabetic ketoacidosis and nonketotic hyperglycemia were treated on a low-dose insulin protocol using intravenous infusion of insulin with the established rate of 0.1U·kg(-1)·h(-1). The patients received intravenous fluids and nutrition orally and intravenously. The levels of serum ICAM-1, E-selectin, and 8-iso-prostaglandin F(2α) (8-iso-PGF(2α)); the activities of superoxide dismutase (SOD); the total antioxidant capacity (TAC) and the contents of malondialdehyde (MDA) in 68 patients with hyperglycemia crisis on admission and after insulin therapy with resolution of hyperglycemia and ketoacidosis (72 h after resolution) were measured. Another 33 healthy individuals served as normal controls. RESULTS: The activities of SOD and TAC at admission were lower in patients with hyperglycemia crisis than in normal controls, and the levels of MDA, 8-iso-PGF(2α), ICAM-1 and E-selectin were higher in patients with hyperglycemia crisis than in normal controls (all p<0.05). The activities of SOD and TAC in patients at resolution were significantly lower than in patients at admission and were significantly higher than in controls (p<0.05). The levels of MDA, 8-iso-PGF(2α), ICAM-1 and E-selectin in patients at resolution were markedly lower than in patients at admission (all p<0.05) and were significantly higher than in normal controls (p<0.05). There was a significant positive correlation between ICAM-1 and SOD (r=0.32, p<0.05) and between E-selectin and MDA (r=0.30, p<0.05) in patients at admission, and the level of E-selectin was positively correlated with MDA and 8-iso-PGF(2α) in patients at resolution (r=0.33, 0.36, p<0.05). In stepwise regression analysis, MDA and 8-iso-PGF(2α) showed a significant association with E-selectin, and 8-iso-PGF(2α) showed a significant association with ICAM-1. CONCLUSION: The oxidative stress and the levels of serum circulating adhesion molecules are significantly changed in patients with hyperglycemia crisis. Intensive insulin therapy can attenuate the abnormity of oxidative stress and the levels of serum circulating adhesion molecules in patients with hyperglycemia crisis.

Correlation between glucose fluctuations and carotid intima-media thickness in type 2 diabetes.

AIMS: To evaluate the relationship between blood glucose fluctuations and carotid atherosclerosis (AS) in type 2 diabetic patients. METHODS: A case-control study included 36 patients with type 2 diabetes and 10 controls. According to the levels of carotid intima-media thickness (CIMT), 36 diabetic patients were classified into two groups, the diabetes mellitus without atherosclerosis (DM-AS, n=20) and diabetes mellitus with atherosclerosis (DM+AS, n=16). The glucose excursions were assessed by the following parameters obtained from continuous glucose monitoring system for 72h: the mean blood glucose (MBG) and its standard deviation (SD), the area under the ROC curve when the blood glucose was higher than 7.8mmol/L (AUC7.8), the mean amplitude of glycemic excursion (MAGE), the mean of daily differences (MODD), and the largest amplitude of glycemic excursion (LAGE). RESULTS: The levels of MBG, SD, MAGE, LAGE, and AUC7.8 were gradually increased with the progression of atherosclerosis (P<0.05). The Spearman's correlation analysis showed that the CIMT was correlated to the age (R=0.58, P<0.001), the duration (R=0.50, P<0.001), the MAGE (R=0.34, P=0.021), and the LAGE (R=0.31, P=0.035). CONCLUSION: These results suggest that glucose fluctuations may accelerate atherogenesis in older type 2 diabetic patients who had a longer duration.

[The relationship between serum level of leptin and oxidative stress in patients with hyperglycemia crisis].

OBJECTIVE: To investigate the relationship between the levels of serum leptin and oxidative stress in patients with hyperglycemia crisis. METHODS: A total of 96 patients with diabetic ketoacidosis (DKA) and nonketotic hyperglycemia (NKH) were treated on a low-dose insulin protocol using intravenous infusion of insulin with the established rate of 0.1 Uxkg(-1)xh(-1), with the patients on intravenous fluids and receiving nutrition by mouth and vein. The levels of serum leptin, 8-iso-prostaglandin F(2 alpha) (8-iso-PGF(2 alpha)), the activities of superoxide dismutase (SOD), total antioxidant capacity (TAC) and the contents of malondialdehyde (MDA) in 96 patients with hyperglycemia crisis on admission and after insulin therapy with resolution of hyperglycemia and ketoacidosis (72 hours) were measured. Another 35 healthy individuals served as normal control. RESULTS: The activities of SOD, TAC and the levels of leptin before treatment were lower in patients with hyperglycemia crisis than in normal controls, and the levels of MDA and 8-iso-PGF(2 alpha) were more markedly elevated than those in normal controls (all P<0.05). The activities of SOD, TAC and the levels of leptin in patients after treatment were significantly higher than those in patients before treatment, and the levels of MDA and 8-iso-PGF(2 alpha) were significantly lower than those in patients on admission (all P<0.05). There was significant positive correlation between leptin and MDA in patients before treatment (r=0.38, P<0.05), and the level of leptin was negatively correlated with MDA and 8-iso-PGF(2 alpha) in patients after treatment (r(1)=-0.35, r(2)=-0.37, both P<0.05). In stepwise regression analysis, MDA and 8-iso-PGF(2 alpha) showed a significant association with leptin. CONCLUSION: The levels of leptin are significantly lowered in patients with hyperglycemia crisis. Oxidative stress may participate in determining the leptin level in hyperglycemia crisis.

[Circulating levels of oxidative stress and adipocytokines in obese subjects].

AIM: To investigate the relationship between adipocytokines levels and oxidative stress in obese males. METHODS: The levels of 8-iso-prostaglandinF2alpha(8-iso-PGF2alpha), superoxide dismutase (SOD), malondialdehyde (MDA), adiponectin, leptin, restistin, TNF-receptors 1(TNF-R1), Interleukin-1beta(IL-1beta) and Interleukin-6(IL-6) were measured in obese men and normal controls. RESULTS: The levels of 8-iso-PGF(2alpha), MDA, leptin, TNF-R1, IL-1beta and IL-6 was significantly higher than that normal controls (P<0.05, P<0.01). The levels of adiponectin and the activity of SOD decreased significantly in obese men.There was no significant difference in the restistin between obese men and normal controls. There was significantly positive correlation between 8-iso-PGF(2alpha) and body mass index (BMI) (r=0.54, P<0.05) in obese. A significantly negative correlation was found between 8-iso-PGF(2alpha) and adiponectin (r=-0.56, P<0.05) in obese subjects. The levels of leptin was negative correlated with body fat content(%)(r=-0.53, P<0.05) in obese subjects. A significant negative correlation was observed between the levels of adiponectin and LDL(r=-0.54, P<0.05), IL-6 (r=-0.41, P<0.05). In a multiple regression analysis model, the levels of adiponectin and IL-6 were the main determinants of the oxidative stress in obese men. CONCLUSION: Changed concentration of adipocytokines was found in obese men. There are significantly correlation with between oxidative stress and adipocytokines.

[The clinical characteristics and its relationship with metabolic syndrome in elderly subjects with impaired glucose regulation].

OBJECTIVE: To study the clinical characteristics of impaired glucose regulation (IGR) in elderly subjects and its relationship with metabolic syndrome (MS). METHODS: The exploration of IGR in 2 810 Chongqing citizens over 40 years old was done by OGTT in a cross-section study. Normal glucose tolerance (NGT), IGR and diabetes (DM) were grouped based on the1999 diagnosis standard of WHO. IGR was composed of impaired fasting glucose (IFG), impaired glucose tolerance (IGT) and both of which. RESULTS: The prevalence of IGR was 18.11%, among which IGT (85.27%). Compared with the NGT group, the IGR group had higher age, body mass index (BMI), blood pressure, triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-c) and HOMA-IR, lower high density lipoprotein cholesterol (HDL-c) and HOMA-B. The IGR group had lower blood pressure, TG and HOMA-IR, and higher HOMA-B than the DM group. When each subgroup of IGR was compared with each other, both IFG plus IGT subgroup and IFG subgroup had higher BMI and HOMA-IR, and lower HOMA-B than IGT subgroup. The prevalences of hypertension, lipid disorder, obesity/overweight, and microalbuminuria in each subgroup of IGR were statistically higher than that of the NGT group. The prevalence of MS in the IFG plus IGT subgroup was higher than that of the IGT subgroup. CONCLUSIONS: The incidence of IGR was high in elderly people over 40 years old in local district of Chongqing city. There were various metabolic disorders in the subgroups of IGR. The IFG plus IGT and IFG group had higher BMI, hypertension, microalbuminuria and HOMA-IR, but lower HOMA-B than the IGT group.