RESUMEN
AIM:To investigate the effect of spermidine(SPD)on pressure overload-induced cardiac hyper-trophy and heart failure model in mice and its underlying mechanisms.METHODS:(1)Eight-week-old male C57BL/6J mice were randomly divided into 4 groups:sham group,sham+SPD group,transverse aortic constriction(TAC)group,and TAC+SPD group.After TAC,the mice in sham+SPD group and TAC+SPD group were fed with 3 mmol/L SPD via drinking water,and the mice in other groups were fed with normal water.Western blot was used to detect the protein ex-pression levels of silent information regulator 6(SIRT6),peroxisome proliferator-activated receptor γ coactivator-1(PGC-1)and mitofusin 2(MFN2).Adult mouse cardiomyocytes were isolated to detect cell length and width.Wheat germ agglu-tinin staining was used to detect the cardiac cell size.Masson staining was used to detect the extent of fibrosis.Echocar-diography was used to detect cardiac function and myocardial hypertrophy.Transmission electron microscopy was used to analyze mitochondrial morphology.Oxygraph-2k high-resolution respirometer was used to detect cardiac mitochondrial oxy-gen consumption.(2)In vitro,primary rat ventricular cardiomyocytes were cultured and treated with angiotensin II(Ang II;1 μmol/L)to construct a hypertrophy model of cardiomyocytes.These cardiomyocytes were divided into control(Con)group,Con+SPD(1 mmol/L)group,Ang II group,Ang II+SPD group and Ang II+SPD+SIRT6 siRNA(siSIRT6)group.Confocal microscopy was used to detect cardiomyocytes area and mitochondrial.RESULTS:(1)Compared with sham group,cardiac function of the mice in TAC group was significantly decreased(P＜0.05),the degree of myocardial hyper-trophy was significantly increased(P＜0.05),and the expression levels of SIRT6,PGC-1 and MFN2 in the myocardial tis-sue were significantly decreased(P＜0.05).Compared with TAC group,the expression levels of SIRT6,PGC-1 and MFN2 in mouse myocardial tissues of TAC+SPD group were significantly increased(P＜0.05),pathological myocardial hy-pertrophy was reduced(P＜0.05),the numbers of mitochondria and mitochondrial cristae were increased(P＜0.05),mito-chondrial function was restored(P＜0.05),myocardial fibrosis was alleviated(P＜0.05),and cardiac function was im-proved(P＜0.05).(2)In vitro,compared with Con group,the expression levels of SIRT6,PGC-1 and MFN2 in cardio-myocytes of Ang II group were decreased(P＜0.05),and the degree of cardiomyocyte hypertrophy was significantly in-creased(P＜0.05).Treatment with SPD increased the expression levels of SIRT6,PGC-1 and MFN2 in cardiomyocytes of Ang II group(P＜0.05),reversed myocardial hypertrophy and improved mitochondrial dynamics(P＜0.05).Compared with Ang II group,the expression levels of SIRT6,PGC-1 and MFN2 in Ang II+SPD+siSIRT6 group showed no significant changes,and the degree of cardiomyocyte hypertrophy and mitochondrial dynamics also had no statistically significant changes.CONCLUSION:Spermidine promotes the expression of SIRT6,PGC-1 and MFN2,thus improving mitochon-drial function,reducing myocardial hypertrophy and alleviating heart failure in mice with pressure overload.
RESUMEN
Objective:To investigate the relationship between intramuscular adipose tissue index (IATI) calculated from computed tomography images at transverse process of the first lumbar and all-cause mortality in maintenance dialysis patients, and to provide a reference for improving the prognosis in these patients.Methods:It was a multicenter retrospective cohort study. The clinical data of patients who received maintenance hemodialysis or peritoneal dialysis treatment from January 1, 2017 to December 31, 2019 in 4 grade Ⅲ hospitals including Zhongda Hospital Affiliated to Southeast University, Taizhou People's Hospital Affiliated to Nanjing Medical University, Affiliated Hospital of Yangzhou University, and the Third Affiliated Hospital of Soochow University were retrospectively collected. IATI was calculated by low attenuation muscle (LAM) density/skeletal muscle density. The receiver-operating characteristic curve was used to determine the optimal cut-off value of IATI, and the patients were divided into high IATI group and low IATI group according to the optimal cut-off value. The differences of baseline clinical data and measurement parameters of the first lumbar level between the two groups were compared. The follow-up ended on December 23, 2022. The endpoint event was defined as all-cause mortality within 3 years. Kaplan-Meier survival curve and log-rank test were used to analyze the survival rates and the differences between the two groups. Multivariate Cox regression analysis models were used to analyze the association between IATI and the risk of all-cause mortality in maintenance dialysis patients. Multivariate logistic regression analysis model was used to analyze the influencing factors of high IATI.Results:A total of 478 patients were eligibly recruited in this study, with age of (53.55±13.19) years old and 319 (66.7%) males, including 365 (76.4%) hemodialysis patients and 113 (23.6%) peritoneal dialysis patients. There were 376 (78.7%) patients in low IATI (<0.42) group and 102 (21.3%) patients in high IATI (≥0.42) group. The proportion of age ≥ 60 years old ( χ2=24.746, P<0.001), proportion of diabetes mellitus ( χ2=5.570, P=0.018), fasting blood glucose ( t=-2.145, P=0.032), LAM density ( t=-3.735, P<0.001), LAM index ( t=-7.072, P<0.001), and LAM area/skeletal muscle area ratio ( Z=-9.630, P<0.001) in high IATI group were all higher than those in low IATI group, while proportion of males ( χ2=11.116, P<0.001), serum albumin ( Z=2.708, P=0.007) and skeletal muscle density ( t=12.380, P<0.001) were lower than those in low IATI group. Kaplan-Meier survival analysis showed that the 3-years overall survival rate of low IATI group was significantly higher than that in high IATI group (Log-rank χ2=19.188, P<0.001). Multivariate Cox regression analysis showed that IATI<0.42 [<0.42/≥0.42, HR(95% CI): 0.50 (0.31-0.83), P=0.007] was an independent protective factor of all-cause mortality, and age ≥60 years old [ HR (95% CI): 2.61 (1.60-4.23), P<0.001], diabetes mellitus [ HR (95% CI): 1.71 (1.06-2.78), P=0.029] and high blood neutrophil/lymphocyte ratio [ HR (95% CI): 1.04 (1.00-1.07), P=0.049] were the independent risk factors of all-cause mortality in maintenance dialysis patients. Stepwise Cox regression analysis showed that IATI<0.42 was still an independent protective factor of all-cause mortality in maintenance dialysis patients [<0.42/≥0.42, HR (95% CI): 0.45 (0.27-0.76), P=0.003]. Multivariate logistic regression analysis showed that low skeletal muscle density [ OR (95% CI): 0.84 (0.81-0.88), P<0.001] and high serum triglyceride [ OR (95% CI): 1.39 (1.07-1.82), P=0.015] were the independent influencing factors of IATI≥0.42. Conclusion:IATI<0.42 of the first lumbar level is an independent protective factor of all-cause mortality in maintenance dialysis patients. Localized myosteatosis within high-quality skeletal muscle may reduce the risk of all-cause mortality in these patients.
RESUMEN
Calciphylaxis is a vascular disease caused by a combination of multiple factors, and the calcified ischemic lesion results in the severe skin damage accompanied by unbearable pain. Calciphylaxis tends to occur in patients with end-stage renal disease, and the treatment of this disease faces enormous challenges. Current treatment recommendations are mainly based on clinical experience and observational research reports, and there is still a lack of clinical practice standards or consensus for managing calciphylaxis. Therefore, this paper will review the effective treatment methods and off-label use of calciphylaxis based on literature reports, providing reference for the clinical treatment of the disease.
RESUMEN
Objective:To observe the efficacy and safety of roxadustat in the treatment of renal anemia in calciphylaxis dialysis patients who had poor response to recombinant human erythropoietin (rHuEPO).Methods:This study was a prospective cohort study. The dialysis patients who were diagnosed with calciphylaxis and had previous regular use of rHuEPO≥3 months with hemoglobin (Hb) levels<110 g/L in the Department of Nephrology of Zhong Da Hospital affiliated to Southeast University from January 1, 2019 to March 28, 2021 were recruited. The effect of oral roxadustat in calciphylaxis dialysis patients with renal anemia was analyzed by self-comparison method.Results:There were totally 18 calciphylaxis dialysis patients with renal anemia enrolled in the study and the age was (49.7±16.2) years old, including 11 males and 7 females, and 14 cases on hemodialysis and 4 cases on peritoneal dialysis. The high-sensitivity C-reactive protein level was 27.3(15.6, 48.5) mg/L(reference value 0-3 mg/L) at baseline. The baseline Hb level was (85.4±11.6) g/L, and after 3 months of oral roxadustat, the Hb level was (105.8±15.2) g/L ( t=-9.282, P<0.001). The Hb compliance rate was 44.4%(8/18). Ferritin decreased significantly at 3 months compared with the baseline level [208.0(59.0, 306.3) μg/L vs 229.0(127.3, 385.2) μg/L, Z=-3.637, P<0.001]. The total iron binding capacity level increased significantly compared with the baseline level [127.0(65.0, 211.5) μmol/L vs 105.5(43.8, 153.7) μmol/L, Z=-2.156, P=0.031]. Transferrin saturation level at 3 months was lower than that at baseline, but there was no significant difference [20.2%(14.2%, 27.7%) vs 20.5%(18.7%, 34.9%), Z=-1.546, P=0.122]. No adverse reactions occurred during the observation period. Conclusion:The application of roxadustat can effectively correct Hb level and improve iron metabolism with high safety in calciphylaxis dialysis patients with renal anemia under inflammatory status.
RESUMEN
Arteriosclerosis is the leading cause of stroke, with a fatality rate surpassing that of ischemic heart disease. High-resolution vessel wall magnetic resonance imaging is generally recognized as a non-invasive and panoramic method for the evaluation of arterial plaque; however, this method requires improved signal-tonoise ratio and scanning speed. Recent advances in high-density head and neck coil arrays are characterized by broad coverage, multiple channels, and closefitting designs. This review analyzes fast magnetic resonance imaging from the perspective of accelerated algorithms for vessel wall imaging and demonstrates the need for effective algorithms for signal acquisition using advanced radiofrequency system. We summarize different phased-array structures under various experimental objectives and equipment conditions, introduce current research results, and propose prospective research studies in the future.
RESUMEN
In order to meet the needs of the flow cytometry for the simultaneous analysis of multiple fluorescence wavelengths and small volume, the design method of flow cytometry spectrum analysis system is presented by analyzing the characteristics of Dyson structure. And according to the method, a flow cytometry spectrum analysis system is disigned with Dyson type.The system's spectral range is 400 nm to 800 nm, the defocused spot size is less than the pixel size 24μ mm, the ransfer function value is above 0.8 at the Nyquist cut-off frequency 21 lp/mm,the spectral resolution is less than 3 nm, and the overall size is 83.54 mm×85.60 mm.The system has good optical performance and small volume, which meets the needs of the flow cytometry fluorescence spectral analysis. The outstanding innovation of this system is the application of Dyson light splitting structure and EMCCD detector which is high speed and high sensitivity.
Asunto(s)
Diseño de Equipo , Citometría de FlujoRESUMEN
Objective To investigate the effects of active vitamin D (VD) on the expression of triggering receptor expressed on myeloid cells-1 (TREM-1) in renal tissue of diabetic nephropathies (DN) rats and to explore the impact of TREM-1 on adhesion and migration capacity of macrophage.Methods DN rat models were established by streptozotocin.Rats were randomly distributed into four groups:control (NC) group,VD group,DN group and DN+VD group (DN rats with 0.1 μg · kg-1 · d-1 calcitriol by garages).Rats were sacrificed respectively at 8 weeks and 12 weeks after treatment.Pathological changes in kidney tissue were detected and the expressions of CD68 and TREM-1 were acquired by immunohistochemistry stain and Western blotting.In vitro,RAW264.7 cells were divided into NC group,VD group,high glucose (HG) group and HG+VD group.In HG+VD group rats were treated by high glucose with 10-8 mol/L 1,25(OH)2D3.TREM-1 expression was measured by immunohistochemistry stain and Western blotting,and the ability of macrophage in migration and adhesion was evaluated by Transwell migration assay and adhesion assay.TREM-1 siRNA was transferred to silence TREM-1 expression,while plasmid of TREM-1 was transferred for high expression.Their ability of adhesion and migration in macrophage and the effect of 1,25(OH)2D3 were examined.Results (1) Compared with the NC group,the expressions of CD68 and TREM-1 were increased in DN group (P ＜ 0.05),whereas markedly decreased in DN+VD group (P ＜ 0.05).(2) The number of adhesion and migration cells,and the expression of TREM-1 protein in macrophage were obviously increased in HG group as compared with those in NC group (all P ＜ 0.05);whereas above changes were markedly decreased in HG+VD group than those in HG group (P ＜ 0.05).(3) The number of adhesion and migrated macrophage was reduced after TREM-1 siRNA intervention (all P ＜ 0.05).VD could significantly decrease the effect of high glucose on adhesion and migrated macrophages after TREM-1 siRNA (all P ＜ 0.05).(4) Adhesion and migration of macrophage were increased via TREM-1 overexpression (all P ＜ 0.05),but the effects of VD on high glucose-induced adhesion and migration of macrophage were disappeared.Conclusions VD can suppress the adhesion and migration of macrophage via reducing the expression of TREM-1,and inhibit infiltration of macrophage in renal tissue of DN rats.
RESUMEN
Based on the new trend of transition from information service to knowledge service currently emerging in the field of information service,the paper presents the construction of a knowledge service system based on clinical data center.It introduces the architecture of the system,analyzes its features,describes its application effect and discusses the issues to be concerned for the further development of the knowledge service system in the future.
RESUMEN
Objective To investigate the effect of macrophages on podocytes apoptosis in diabetic nephropathy. Methods Differentiated mouse macrophages (RAW264.7) were exposed to normal glucose, high glucose, then the conditioned media (CM) was collected and considered as NC-CM or HG-CM, respectively. Western blotting and immunofluorescent staining were used to detect the specific markers for M1 macrophages (iNOS) and M2 macrophages (MR). ELISA was used to detect the concentration of TNF-α in the CM. Then normal PRMI 1640 media (control), NC-CM or HG-CM was added to podocytes. In some experiments, ROS inhibitors (Tempo), p38 MAPK inhibitor (SB203580), anti-TNF-α neutralizing antibody, and IgG1 isotype control were respectively added to cells with HG-CM. Besides, recombinant mouse TNF-α alone was applied to incubate podocytes. Podocytes apoptosis was accessed by Annexin V-FITC/PI and Hoechst33342 staining. DCFH-DA staining was used to analyse ROS level. Western blotting was used to detect cleaved casepase-3, p38MAPK, and p-p38MAPK protein. Results Macrophages were activated when exposed to high glucose, displaying pro-inflammatory M1 polarization with higher iNOS and lower MR expression. HG-CM but not NC-CM trigged podocytes apoptosis, up-regulated ROS, cleaved casepase-3 and p-p38MAPK. However, the podocytes apoptosis trigged by HG-CM was abolished by either a ROS inhibitor (Tempo) or a p38 MAPK inhibitor (SB203580). Additionally, TNF-α was increased in the HG-CM. TNF-α protein in macrophage was aslo increased when exposed to high glucose. Anti-TNF-α neutralizing antibody blunted the apoptotic response, excess ROS generation and p-p38 MPAK expression in podocytes induced by HG-CM. Moreover, addition of recombinant TNF-α similarly led to podocytes apoptosis, increased ROS and p38 MPAK expression. Conclusion M1 macrophages activated by high glucose released TNF-α to promote podocytes apoptosis via ROS-p38 MAPK pathway.
RESUMEN
Objective To investigate the effect of 1,25(OH)2D3 on high glucose induced podocyte injury and its signal transduction mechanism.Methods Differentiated mouse podocytes were exposed to normal glucose,high glucose,and different concentrations of 1,25(OH)2D3 or LY294002 (a selective PI3K inhibitor) for 24 h.PCR and immunofluorescent staining were used to detect nephrin,podocin,and desmin.Western blotting was used to detect protein expression of nephrin,podocin,desmin,PI3K,Akt and p-Akt.Results Compared with high glucose group,1,25(OH)2D3 (100 nmol/L and 1000 nmol/L) significantly up-regulated the expression of podocin and nephrin in podocytes induced by high glucose (P ＜ 0.05).Meanwhile,1,25(OH)2D3 (100 nmol/L) significantly reduced the expression of desmin (P ＜ 0.05).PI3K and p-Akt were obviously reduced in high glucose group.In the presence of 1,25(OH)2D3,the trends were reversed.However the above effects of 1,25(OH)2D3 were abolished when p-Akt was blocked by the PI3K inhibitor LY294002.Conclusions 1,25 (OH)2D3 can inhibit high glucose-induced pedocyte injury through PI3K/p-Akt signaling pathway.
RESUMEN
Objective To explore the safety and feasibility of guiding catheter passing through spasmodic vessels in patients undergoing percutaneous coronary intervention (PCI) via radial artery access by the aid of PCI guiding wire and balloon .Methods The clinical data of 33 coronary artery disease (CAD) patients undergoing PCI via radial artery access with radial artery or (and) brachial artery spasm ( group A ) were retrospectively analyzed .Among all these patients , guiding catheters were delivered through the spasmodic vessels successfully by the aid of PCI guiding wires and balloons .The clinical data of other 38 CAD patients having PCI during the same period performed by other operators via radial artery or ( and ) brachial artery approach and experienced vessel spasm were anlysed as the control ( group B ) .All patients in group B received conventional anti-spasm management during PCI .All vessel spasm was identified by angiography.For patients in group A , a diameter of 0.014 inch guiding wire was chosen to pass through the spasmodic vessel segment carefully and gently .The diameter of balloon should be chosen according to the diameter of guiding catheter .A balloon diameter of 2.0 mm and 2.5 mm was corresponded to 6F and 7F guiding catheter respectively .The balloon was advanced to the tip of guiding catheter , keeping a half in catheter and a half in vessel followed by inflating the balloon with a pressure of 8 atm.The balloon was kept inflated the guiding catheter was pushed in vitro carefully and slowly until the catheter passed through the spasmodic vessel segment .Then the balloon was deflated and pulled out together with PCI guiding wire . Exchanged a diameter of 0.035 inch wire and completed the positioning of guiding catheter .After finishing the PCI, radial or ( and) brachial angiography was performed again to observe if spasm disappeared and to determine if there any contrast medium exudation .For patients in group B , routine approach was applied including administration of nitroglycerine , diltiazem or nitroprusside etc . to relieve vessel spasm. Results The location of vessel spasm was similar in group A and group B ( P=0.150 ) , and the incidence rate of spasm in brachial artery was higher than that in radial artery in both groups .The chance of guiding catheter crossing the spasmodic vessel segment was significantly higher in group A than in group B ( 100%vs.39.5%, P=0.00).In patients whose guiding catheter could pass through the spasmodic vessel segment successfully , time spent in group A was shorter than in group B ( P=0.000 ) .The patient number which time spent was less than five minutes , five to 15 minutes and more than 15 minutes was 30 and 2 ( 90.1%vs.13.3%) , 3 and 7 ( 9.9% vs.46.7%) and 0 and 6 ( 0% vs.40.0%) in group A and in group B respectively.The incidence of forearm hematoma was lower in group A than in group B without statistical difference [6.1%(2/33) vs.18.4%(7/38), P =0.113].Conclusions It is safe and feasible for passing guiding catheter through spasmodic vessels during PCI via radial artery access by the aid of PCI guiding wire and balloon .
RESUMEN
Diabetic nephropathy is caused by manifold factors .Increasing evidences suggest that immunoregulation abnormality caused by inflammatory cell infiltration and proinflammatory factor overexpression in kidney tissue has a key role in the occurrence and development of diabetic nephropathy .As a novel immunomodulator , activated vitamin D 3 could inhibit inflammatory factor and regulate immunity response .The author reviews the effect of activated vitamin D 3 related to diabetic nephropathy , from the pointviews of immu-nocyte, cytokines and rennin-angiotensin system.The roles of macrophage, T lymphocytes, dendritic cell, transforming growth factor-β1, and rennin-angiotensin system in kidney injury and immunal regulation of activated vitamin D 3 are discussed.
RESUMEN
Objective To investigate the effect of 1,25(OH)2D3 on high glucose induced macrophage activation and its underlying signal transduction mechanism.Methods RAW 264.7 cells were used to perform cell culture,the activity of intracellular iNOS was measured.VDR siRNA and PPARγ antagonist pre-treatment with macrophages were done before using 10-8 mol/L1,25(OH)2D3 to intervene high glucose pre-incubated macrophages.M1 markers including iNOS,TNF-α,IL-12,M2 markers including MR,Arg-1,IL-10 and nuclear receptors VDR and PPARγ were separately examined.Results The iNOS activity was increased in a glucose-dose and time dependent manner.Particularly,25 mmol/L glucose at 24 h gave the maximum response.After being treated with 25 mmol/L glucose for 24 h,not only inflammatory cytokines of TNF-α,IL-12 in the supernatant were increased,but quantitative real-time PCR and Western blotting analysis showed iNOS was also up-regulated (P ＜ 0.05).However,M2 markers,i.e.MR and Arg-l were significantly decreased (P ＜ 0.05).When in the presence of 1,25(OH),D3,the trends were reversed:the markers of M1,including TNF-α,IL-12 and iNOS were obviously reduced (P ＜ 0.05),while M2 markers,IL-10,Arg-1 and MR were increased (P ＜ 0.05).In addition,VDR and PPARγ were also increased (P ＜ 0.05).However,the above effects of 1,25 (OH)2D3 were abolished when further inhibited the expression of VDR and PPARγby VDR siRNA and PPARγ antagonist.Besides,accompanied by VDR,PPARγwas also decreased upon the treatment with VDR siRNA (P ＜ 0.05).Conclusion 1,25(OH)2D3 can promote high glucose induced classically activated macrophages (M1) converting to alternatively activated macrophages (M2) and this is achieved through VDR-PPARγ pathway.
RESUMEN
Objective To investigate the clinical effect of basilic vein transposition arteriovenous fistula in hemodialysis patients.Methods NInety patients with maintenance hemodialysis received in the NO.180 Hospital of the Chinese People's Liberation Army from September 2011 to September 2012 were randomly divided into two groups,and each group of 45 cases.Patients in the observation were given brachial-basilica transposition arteriovenous fistula,while patients in the control group received artificial vascular graft arteriovenous fistula.The fistula maturation time,dialysis blood flow,urea removal index (Kt/V),patency rates and complications were respectively recorded.Results Compared with control group,fistula maturation time,dialysis blood flow,Kt/V and complications rates in observation group were significantly higher((14.4±3.2)weeks vs.(16.1±2.7) weeks,(291.5±33.9) ml/min vs.(252.6±29.8) ml/min,(1.6±0.2) vs.(1.3±0.3);t =4.538,3.984,4.016;P＜ 0.05).Complications (ipsilateral upper limb swelling,thrombosis,venous ectasia and arteriovenous fistula stenosis) incidence were significantly lower than those of control group (2.2％vs.13.3％,2.2％ vs.11.1％,6.6％ vs.17.8％,11.1％ vs.24.4％;x2=5.463,4.972,5.017,3.968;P ＜0.05).Patency rates of observation group in 3 months,6 months,1 year and 2 years were also significantly higher than those of control group (97.8％ vs.93.3％,91.1％ vs.84.4％,88.9％ vs.75.6％,84.4％ vs.68.9％,x2 =5.315,4.238,7.024,5.913;P＜0.05).Conclusion Basilic vein transposition arteriovenous fistula is reliable and effective for hemodialysis patients.It can achieve adequate dialysis and less complications and It is worth of clinical application.
RESUMEN
Objective To investigate the relationship between the prognosis of patients with coronary heart disease in elderly heart failure between lipid levels .Methods 720 cases of coronary heart disease in elderly patients with heart failure ,according to the survival of the follow-up period were divided into survival group and death group , comparing lipid levels in each group of patients , the use of multivariate Cox regression analysis to investigate the effects of lipids in elderly patients with heart failure ,coronary artery disease prognosis .Results Age group of death , old myocardial infarction ( OMI ) , the probability of occurrence of atrial fibrillation , anemia was significantly higher than that of the survival group (χ2 =5.626,4.597,5.632,6.461,all P<0.05),HDL-C,EF,ACEI/ARB and statins was significantly lower than those of the survival group (t=10.725,9.563,8.457,9.170,P<0.05 or P<0.01).An-giotensin-converting enzyme inhibitors ( ACEI)/angiotensin Ⅱreceptor antagonists ( ARB) would help to improve the prognosis(r=0.678,P<0.01),whereas low levels and low levels of HDL-C,NYHAⅢ-grade Ⅳ,age was a risk fac-tor(r=1.052,2.298,2.586,all P<0.01).Conclusion There is a close contact between HDL-C and prognosis of heart failure in elderly patients with coronary heart disease ,in order to improve the prognosis of patients ,it can be ap-propriately managed to elevate HDL-C levels.
RESUMEN
Objective To cultivate mice glomerular podocyte with high glucose and high Ang-Ⅱ,observe the morphocytology of podocyte,and investigate protein kinase C (PKC) activity,the expressions of nephrin and CD2-associated protein (CD2AP),and apoptosis of podocyte with injuries cultivations.Methods The conditions immortalization mice podocyte cell lines was cultivated,after passage and differential induce,the cell lines was divided into six groups:normal group,high glucose group,high Ang-Ⅱ] group,the above two mixed group,intervened group of PKC inhibitor,and hypertonic group.After 72 hours,cell apoptosis rate was detected with flow cytometer; PKC activity was detected with enzyme linked immunosorbent assay (ELISA) in 24 hours and 48 hours ; The expressions of nephrin and CD2AP mRNA were detected by real time-polymerase chain reaction (RT-PCR).Results (1)After 24 hours,the PKC of high glucose group and high Ang-Ⅱ group were more activated than normal group [(47.09 ± 1.19) pmol/L,(42.93 ±0.71) pmol/L,P ＜0.05].The activated phenomenon was more obvious in the mixed group.Compared to 24 hours [(58.75 ±0.71) pmol/ L,P ＜ 0.01],PKC was more activated in 48 hours (P ＜ 0.05).(2) Nephrin mRNA was reduced in podocyte exposed to high glucose and high Ang-Ⅱ at least 24 hours (56.87 ± 0.74,48.54 ± 0.86,P ＜ 0.05),and was reduced more in mixed group (11.7 ± 1.54,P ＜ 0.01).(3) CD2AP mRNA was reduced in podocyte exposed to high glucose,high Ang-Ⅱ,and mixed environment at least 48 hours (56.09 ± 1.46,67.68 ±2.58,and 54.08 ±2.74,P ＜0.05).The decrease trend of nephrin and CD2AP mRNA was restrained by PKC inhibitor(90.75 ± 1.33,P ＜0.05).(4) After 48 hours and 72 hours,the apoptosis rates of high glucose group,high Ang-Ⅱ group,and mixed group were risen compared to normal group (P ＜ 0.05).The increase of apoptosis rate can be restrained by PKC inhibitor.Conclusions High glucose,and high Ang-Ⅱ can activate the podocytes PKC activity,inhibit the expression of nephrin and CD2AP mRNAs,and induce podocyte apoptosis.PKC signaling pathway plays an important role in the injury mechanism of Ang-Ⅱ and high glucose to mice podocyte.
RESUMEN
Objective To investigate the effects and underlying mechanism of calcitriol on ameliorating podocytes impairment in DN rats.Methods SD rats were randomly divided into four groups:normal control (NC) group,calcitriol treatment (VD) group:calcitriol 0.1μg· kg--1 d-1,diabetic nephropathy (DN) group:streptozocin (STZ) 58 mg/kg,DN treated with calcitriol (DN + VD) group:calcitriol 0.1 μg · kg-1 · d-1 + STZ 58 mg/kg.Rats were sacrificed at the end of 18 weeks.Results Compared with the DN group,the DN + VD group exhibited significantly lower proteinuria by 36％,improved renal histology at the end of the experiment (P ＜ 0.05),and similar levels of blood glucose,serum urea nitrogen as well as body weight (P ＞ 0.05).There were no significant differences in the serum concentrations of creatinine,calcium and phosphorus among the four groups (P ＞ 0.05).In DN group,the expressions of nephrin,podocin,VDR,PI3K-p85 and p-Akt were significantly decreased and the expression of desmin was increased compared to NC group.Calcitriol treatment could attenuate the above changes.Additionally,a positive correlation was observed between the expressions of nephrin and VDR (r=0.776,P ＜ 0.05).Likewise,the expression of nephrin was positively correlated with either PI3K -p85 or p-Akt (r=-0.736,r=0.855,all P ＜ 0.05).Conclusion Calcitriol can ameliorate podocytes injury in DN rats,which might be related with the further up-regulation of PI3K/p-Akt signaling pathway.
RESUMEN
Objective To investigate the effect of active vitamin D (VD) on macrophage M1 and M2 phenotype and its role in protecting podocyte impairment in diabetic nephropathy (DN).Methods Diabetes mellitus rats were established by intraperitoneal injection with streptozocin.Rats were randomly divided into four groups:normal-1 (NC-1,n=8),normal-2 (NC-2,n=8,normal rats treated with calcitriol 0.1 μg· kg-1 · d-1 by gavages),DN (n=24) and VD (n=24,DN+calcitriol 0.1 μg· kg-1 · d-1 by gavages).Blood glucose and body weight were assessed,and 24-hour urine was collected regularly.Blood and urine samples were taken for biochemical study,and kidney tissues were used for PAS staining to assess histological changes.Immunohistochemical staining was used to detect number of CD68 + macrophage.Western blotting was used to detect protein expressions of nephrin,podocin,CD68,M1 specific marker of inducible nitric oxide synthase (iNOS),TNF-α and M2 specific marker of CD163,arginase 1 (Arg-1),mannose receptor (MR).Results (1) In DN group,levels of BUN,Scr,urinary protein and glomerular mesangial matrix proliferation were significantly higher (P ＜ 0.05),and the expressions of nephrin,podocin were significantly decreased compared with NC groups (P ＜ 0.05).These above changes were significantly improved in VD group (P ＜ 0.05).(2)Number of CD68 + macrophage infiltration in DN group was increased in a time dependent manner compared with NC groups,which was significantly reduced in VD group (P ＜ 0.05).(3)To further definite M1 and M2 macrophage activation phenotype,the protein expressions of iNOS and TNF-α was increased in DN group at 8th,14th,18th weeks compared with NC groups (P ＜ 0.05),which were significantly decreased in VD group (P ＜ 0.05).Although,there were no significant difference of protein expressions of CD163,Arg-1 and MR between VD and DN group at both 8th and 14th week (P ＞ 0.05),the protein expressions of CD163,Arg-1 and MR were higher in VD group at 18th week than that in DN group (P ＜ 0.05),and the ratio of CD163/CD68 was also enhanced in VD group (P ＜0.05).(4)Moreover,the protein expression of iNOS was negatively correlated with expression of either nephrin or podocin (r =-0.707,P ＜ 0.01; r =-0.712,P ＜ 0.01),whereas the protein expression of CD163 was positively correlated with expression of either nephrin or podocin (r =0.627,P＜ 0.01; r=0.613,P ＜ 0.01).Conclusion Vitamin D can regulate macrophage phenotype,via inhibiting M 1 macrophage activation and enhancing M2 macrophage activation to protect podocyte impairment.
RESUMEN
Objective To explore whether high glucose (HG)-induced endothelial-to-mesenchymal transition (EndMT) could be transitioned into mesenchymal stem cells (MSCs) and further differentiated into chondrocytes.Methods Human aortic endothelial cells (HAECs) were divided into three groups:normal glucose (NG,5.5 mmol/L glucose) group,HG (30 mmol/L glucose) group,and mannitol (5.5 mmol/L glucose + 24.5 mmol/L mannitol) group,and were cultured for 48 h.Immunofluorescence staining was performed to detect the co-expression of CD31 (endothelial markers),and fibroblast-specific protein 1 (FSP1,fibroblast markers).The expression of CD31 and FSP1 mRNA and protein was detected by real-time PCR and Western blotting.When endothelial-derived MSCs were grown in MSC medium for one week,the expression of the MSCs markers CD44,CD10 and the chondrocyte marker SOX9 was detected by Western blotting and RT-PCR.Chondrocyte expression was detected by alcian blue staining.Calcium deposit was analyzed by alizarin red staining.Pathological changes were investigated using electron microscopy.Results The expression of FSP1 mRNA and protein was significantly increased,but the expression of CD31 mRNA and protein was decreased (P ＜0.01),and the cells undergoing EndMT also significantly expressed CD10,CD44 and SOX9 in the HG group compared with those in normal glucose group (P ＜ 0.01).The incubation of HAECs exposed to HG resulted in a fibroblast-like phenotype,wherein increased microfilamentation and a roughened endoplasmic reticulum structure were observed in the cytoplasm.Double staining of the HAECs indicated a co-localization of CD31 and FSP1.After one week culture for chondrocyte medium,the expression of MSCs marker STRO-1 was significantly increased by immunofluorescence staining.Additionally,aleian blue staining in the HG group was positive compared to the NG group.Consistent with the elevation of SOX9 expression,calcium deposit also enhanced in the HG group.Conclusion HG can induce endothelial cells transdifferentiation into chondrocyte-like cells via the EndMT.
RESUMEN
Objective To investigate whether low density lipoprotein receptor (LDLr) pathway involves in the progression of vascular calcification (VC) in hemodialysis patients under microinflammation.Methods Twenty-eight hemodialysis patients were divided into control and inflammation group according to plasma C-reactive protein level.Surgically removed tissues from radial artery of patients receiving arteriovenostomy were used in experiments.Foam cell formation and calcification deposition were observed by hematoxylin-eosin (HE) and alizarin red S staining respectively.VC-related protein expression,such as bone morphogenetic proteins-2 (BMP-2),collagen Ⅰ,alkaline phosphatase (ALP),and LDLr and its related nuclear factor of transcriptional regulation,such as sterol regulatory element binding protein-2 (SREBP-2) and SREBP cleavage-activating protein (SCAP),were detected by immunohistochemistry and immunofluorescence staining.Results HE and alizarin red S staining showed that there were parallel increased foam cell formation and calcium deposit in continuous cross-sections of radial arteries in inflammation group compared to control group,which were closely correlated with increased protein expressions of LDLr,SREBP-2,BMP-2,and collagen Ⅰ as shown by immunohistochemical and immunofluorescent staining.Confocal microscopy confirmed that inflammation enhanced the translocation of SCAP/SREBP-2 complex from endoplasmic reticulum to Golgi,thereby activating LDLr gene transcription.Inflammation increased protein expression of ALP and reduced protein expression of alpha-smooth muscle actin,contributing to the phenotype conversion of vascular smooth muscle cells in calcified vessels from the fibroblastic to the osteogenic,which were the main cell components in VC.Further analysis showed that the disruption of LDLr pathway induced by inflammation was positively correlated with the enhanced expression of BMP-2 and collagen Ⅰ (r=0.782,P＜0.01; r=0.644,P＜0.05).Conclusion Inflammation accelerates the progression of VC in hemodialysis patients through the disruption of LDLr feedback regulation.