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Objective:To investigate the significance of serum fibroblast growth factor 19 (FGF19), Klotho and fibroblast growth factor receptor 4 (FGFR4) protein expression in patients with primary biliary cirrhosis (PBC).Methods:Sixty-three patients with PBC who received treatment in Ningbo Huamei Hospital, University of Chinese Academy of Sciences between August 2017 and July 2020 were included in the PBC group. An additional 51 healthy patients who concurrently received physical examination in the same hospital were included in the control group. Serum FGF19, Klotho and FGFR4 protein expression were determined by enzyme linked immunosorbent assay.Results:Serum FGF19 and FGFR4 protein in PBC group were (178.86 ± 21.28) ng/L and (2.96 ± 0.47) ng/L, respectively, which were significantly higher than those in the control group [(69.93 ± 12.12) ng/L, (1.21 ± 0.35) ng/L, t = 32.51, 27.98, both P < 0.05]. Klotho protein expression in the PBC group was significantly lower than that in the control group [(3.25 ± 0.89) μg/L vs. (9.67 ± 1.53) μg/L, t = 22.08, P < 0.05]. Serum levels of alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase in the PBC group were (84.25 ± 13.24) U/L, (71.82 ± 10.35) U/L, (278.93 ± 32.45) U/L, respectively, which were significantly higher than those in the control group [(23.76 ± 3.42) U/L, (23.10 ± 4.53) U/L, (76.81 ± 16.36) U/L, t = 31.75, 31.26, 40.48, all P < 0.05]. The receiver operating characteristic curve analysis showed that the sensitivity and specificity of FGF19 in the diagnosis of PBC were 76.67% and 61.90%, respectively, they were 58.82% and 66.67% for Klotho protein diagnosis, 54.55% and 76.67% for FGFR4 protein. Pearson analysis revealed that there was a positive linear relationship between FGF19 and FGFR4 protein ( r = 0.78, P < 0.05), while there was a negative linear relationship between Klotho protein and FGFR4 protein ( r = -0.72, P < 0.05). Conclusion:In patients with PBC, serum FGF19 and FGFR4 protein levels are increased, while Klotho protein level is decreased. There is a positive linear relationship between FGF19 and FGFR4 protein, and there is a negative linear relationship between Klotho protein and FGFR4 protein. This study is highly innovative and scientific.
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Objective:To explore the effect of combining hyperbaric oxygen (HBO) therapy with cognition training for persons with vascular cognitive impairment (VCI).Methods:Forty-two persons with VCI were randomly divided into a control group of 19 and a research group of 23. In addition to basic treatment, the control group was given hyperbaric oxygen therapy once a day, 5 days per week for 4 weeks, while the research group received cognition training along with the hyperbaric oxygen therapy. Each person′s cognition was assessed using the Simple Mental Status Scale (MMSE) before and after the four-week treatment. Meanwhile, 3ml of venous blood was collected before eating in the morning to test the plasma levels of Klotho protein and homocysteine using enzyme-linked immunosorbent assays.Results:After the treatment the average MMSE score had improved significantly in both group, with the improvement in the research group′s average significantly greater than that in the control group. The average plasma levels of Klotho protein and homocysteine had also improved significantly more in the research group. In the control group, the only significant improvement was in the average homocysteine level.Conclusions:Hyperbaric oxygen therapy can be an effective supplement to cognition training for persons with vascular cognitive impairment.
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Chronic kidney disease (CKD) is a progressive systemic disease, which is very common in children with kidney disease.With the progress of the disease, patients gradually develop aging-like manifestations, such as se-condary hyperparathyroidism, renal osteodystrophy, ectopic calcification, cardiovascular disease, growth retardation, renal fibrosis and so on. Klotho gene is closely related to human aging.Klotho protein is highly expressed in kidney, which is a key regulator of mineral and bone metabolism in CKD, and has many functions, such as anti-aging, anti-vascular calcification, regulating growth and development, anti-fibrosis and so on.In the early stage of CKD, the levels of Klotho protein in blood and urine decreased.Supplementation of exogenous Klotho protein or activation of endogenous Klotho protein can alleviate renal fibrosis, improve mineral and bone metabolism, reduce vascular calcification and delay the progression of CKD.The basic characteristics of Klotho gene and protein, the biological function of Klotho protein and its role in vascular calcification, CKD mineral osteopathy and the growth and development of children with CKD are reviewed in this article.
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Objective:To observe the effects of modified Liuwei Dihuangtang on serum fibroblast growth factor 23 （FGF23）， full-length intact parathyroid hormone （iPTH）， and 1，25-dihydroxyvitamin D<sub>3 </sub>［1，25（OH）<sub>2</sub>D<sub>3</sub>］ levels and Klotho and FGF23 protein expression in renal and bone tissues of rats exposed to high phosphorus combined with adenine， so as to explore the mechanism of modified Liuwei Dihuangtang against renal osteopathy. Method:One hundred and thirty healthy adult SD rats were randomly divided into five groups， namely normal group（<italic>n</italic>=10），high phosphorus group（<italic>n</italic>=30），model group（<italic>n</italic>=30），modified Liuwei Dihuangtang group（<italic>n</italic>=30） ， and calcitriol group（<italic>n</italic>=30），and rats in each group were further classified based on three time points， namely 8，10， and 12 weeks. Rats in the normal group were fed with normal diet， the ones in the high phosphorus group with high phosphorus diet， and those in the other groups with adenine and high phosphorus diet for inducing renal osteopathy. Rats in the normal group，high phosphorus group， and model group were intragastrically administered with distilled water （10 mL·kg<sup>-1</sup>·d<sup>-1</sup>），the ones in the modified Liuwei Dihuangtang group with modified Liuwei Dihuangtang （2.556 g·kg<sup>-1</sup>·d<sup>-1</sup>） ， and those in the calcitriol group with calcitriol （0.09 μg·kg<sup>-1</sup>·d<sup>-1</sup>）. Result:Compared with the normal group and high phosphorus group at the weeks of 8，10 and 12，the model group displayed significantly elevated blood urea nitrogen（BUN），serum creatinine（SCr），serum phosphorus，iPTH，FGF23，renal interstitial fibrosis score， and FGF23 expression in renal and bone tissues， but lowered serum calcium and 1，25（OH）<sub>2</sub>D<sub>3</sub> and Klotho protein expression in renal and bone tissues（<italic>P</italic><0.05 ，<italic>P</italic><0.01）. Compared with the model group at the weeks of 8，10 and 12， the modified Liuwei Dihuangtang and calcitriol both significantly decreased the serum BUN，SCr，serum phosphorus，iPTH， FGF23， tubulointerstitial semi-quantitative score， and FGF23 expression in renal and bone tissues， while increased the serum calcium，1，25（OH）<sub>2</sub>D<sub>3</sub>， and Klotho protein expression in renal and bone tissues （<italic>P</italic><0.05，<italic>P</italic><0.01）. There was no significant difference in the above-mentioned indexes between the modified Liuwei Dihuangtang group and the calcitriol group at the same time point. Conclusion:Klotho-FGF23 axis is probably involved in renal osteopathy. The modified Liuwei Dihuangtang effectively improves renal function，alleviates pathological changes in renal and bone tissues，and regulates calcium and phosphorus metabolism to protect the bone， which is related to its regulation of Klotho-FGF23 axis.
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Objective: To evaluate the effects of captopril on Klotho protein and related inflammatory factors in the model of hypertensive SD rats. Methods: Eighteen SD rats were randomly divided into normal group, hypertensive model group and captopril experimental group, respectively. N'-nitro-L-arginine (L-NNA) was used by gavage in SD rats to make the hypertensive model. After successful establishment of the model, the experimental group was given captopril for treatment of hypertension, and the normal group and hypertension group were given an equal volume of normal saline by gavage in the SD rats. Blood pressure changes were measured by tail arteries of the SD rats. Enzyme-linked immunosorbent assay (ELISA) was used to measure plasma levels of Klotho protein, thromboxane (TXA2), and endothelin (ET). The level of nitric oxide (NO) was measured by nitrate reductase method. The content of malondialdehyde (MDA) was measured by TBA method, and the activity of superoxide dismutase (SOD) was measured by WST-1 method. The pathological changes of HE stained vascular wall in thoracic aorta of each group were observed. The vascular endothelial cell injury was evaluated by measuring the expression of CD31 by immunohistochemistry (IHC). Results: Compared with those in the normal group, the contents of TXA2, ET and MDA in the model group were significantly higher (P<0.05), while the content of Klothoprotein and NO and SOD activities were lower (P<0.05). Compared with those in the model group, the contents of TXA2, ET and MDA in the captopril group were significantly decreased (P<0.05), and the Klotho protein content, NO content and SOD activity were higher (P<0.05). The results of HE staining showed that the aortic vascular fibrosis and disorder in the model group and the aorticl wall were thicker than those in the normal group. The vessel wall was significantly thinner in the treatment group than in the model group. The results of IHC showed that the aortic endothelial cells in the model group were damaged and shedding. The vascular endothelial cells in the controlled group were increased and gradually repaired. Conclusion: Captopril can reduce the stress response of endothelial cells and repair the vascular endothelium, which can improve vascular endothelial function and increase Klotho protein level.
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Klotho(KL)protein is a membrane-bound protein mainly expressed in the kidney and brain.It can regulate the expression of multiple signal pathways and target genes, such as ion channels, transforming growth factor(TGF), and exert anti-aging effects, kidney protection, anti-tumor and regulating metabolism.Recent studies have found that KL protein plays an important role in renal diseases such as renal interstitial fibrosis(RIF). KL deficiency can induce and promote the progress of RIF, and KL overexpression or supplementation can prevent RIF.This review focuses on the anti-RIF effect of KL protein through multiple signaling pathways, and aims to provide a new direction for anti-RIF therapy.
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@#Objective: To study the effect of anti-aging Klotho protein on immune escape mediated by regulatory T cells (Treg)/helper T cells 17 (TH17) in mice bearing cervical cancer and its mechanism. Methods: The model of cervical cancer-bearing mice were established, and the control group (normal mice), model group (cervical cancer-bearing mice model), and Klotho treatment group (cervical cancer-bearing mice treated with Klotho protein, 200 ng/d) were set up. The weight of cervical cancer tumors in mice of each group was weighed at 7 and 14 days after treatment respectively, PBMCs were separated at the same time. Flow cytometry was used to detect the changes of T lymphocyte function and the proportion of Treg and TH17 cells in mice. qPCR was used to detect the expressions of Foxp3 and RORγt, the key transcription factors of Treg/TH17 cells, in PBMCs of mice in each group. The changes of IL-17, IL-6, IL10, TGF-β and IL-23 in PBMCs were detected by ELISA. The protein expressions of Klotho, TGF-β, Foxp3 and RORγt in PBMCs of mice were detected by WB assay. Results: On the 14th day, the tumor inhibition rate of the cervical cancer-bearing mice in the Klotho group was significantly higher than that in the Model group [(52.16±8.25)% vs (23.33±6.29)% the model group to be supplemented, P< 0.05). Compared with the Control group, the ratios of Treg and TH17 cells in the lymphocytes of the tumor-bearing mice significantly increased (all P<0.05), the ratios of total T lymphocytes (CD3+), auxiliary/induced T lymphocytes (CD3+CD4+) and immune index (CD3+CD4+/CD3+CD8+ cells) decreased significantly (all P<0.05); in addition, the mRNAexpressions of Foxp3 and RORγt genes, cytokines of IL-17, IL-6, IL-10, TGF-β and IL-23, as well as protein expressions of TGF-β1, Foxp3 and RORγt increased significantly (all P <0.05), while the level of Klotho protein significantly decreased in Model group (P<0.05). Compared with the Model group, the above indicators showed opposite changes in Klotho group (P<0.05), but there was no significant difference with the Control group (all P> 0.05). Conclusion: Klotho protein may inhibit Treg/TH17 cell-mediated immune evasion in cervical cancer-bearing mice by inhibiting TGF-β1/Foxp3/RORγt signaling pathway and exert anti-tumor effect.
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Objective To explore the changes and significance of Klotho protein levels in different stages of chronic kidney disease (CKD). Methods From March 2015 to December 2017,176 patients with CKD admitted to nephrology department of Inner Mongolia People＇s Hospital were selected as the study object (CKD group), and 80 healthy patients in our hospital were selected as the control group in the same period. The serum Klotho levels of CKD patients and control group at different stages were detected by double antibody sandwich ELISA, and the differences between each group were compared. Results The serum Klotho level of CKD group (( 4. 84 ± 1. 87) μg／L) was significantly lower than that of the control group ((9. 11± 3. 14) μg／L) ( t＝ 13. 82, P＜0. 01) . One-way anova showed that estimated renal glomerular filtration rate (eGFR),serum albumin (ALB),hemoglobin ( Hb),blood calcium ( Ca) and serum Klotho were gradually decreased,while phosphorus (P) and creatinine (Cr) in serum were gradually increased,and the difference was statistically significant among the five stages( all P＜0. 01). Spearman correlation analysis showed that Klotho level was positively correlated with eGFR and Ca,and negatively correlated with CKD stage,Cr and P (r＝0. 369,0. 160,－0. 200,－0. 250,－0. 230,all P＜0. 05). The multiple linear regression equation showed that Klotho level was positively and independently correlated with eGFR ( t＝ 3. 89, P＜0. 001),and negatively correlated with CKD staging independently (t＝－4. 12,P＜0. 001). Conclusion The expression level of serum Klotho protein in patients with CKD is lower than that of healthy people,and it decreases with the increase of CKD stages,which is closely related to the deterioration of renal function. It can be used as a reference index to evaluate the incidence and severity of CKD.
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Objective? To? investigate? the? protective? effects? of? Klotho? protein,? a? kind? of? single-pass?transmembrane?protein,?on?acute?kidney?injury?(AKI)?in?septic?mice?and?its?mechanism.? Methods? Sixty?SPF?healthy?male?C57BL/6?mice?(6-8?weeks)?were?randomly?divided?into?sham?operation?group?(Sham?group),?sepsis?model?group?(CLP?group)?and?Klotho?protein?injection?group?(CLP+KL?group),?with?20?in?each?group.?The?septic?AKI?mice?model?was?established?by?cecal?ligation?and?puncture?(CLP);?Sham?group?had?the?same?procedure?except?that?the?cecal?was?not?ligated.?The?CLP+KL?group?was?received?Klotho?protein?(0.02?mg/kg)?by?intraperitoneal?consecutive?injection?for?4?days?after?operation;?Sham?group?and?CLP?group?were?injected?with?the?same?amount?of?saline.?Blood?samples?were?obtained?at?24?hours?after?operation,?the?levels?of?serum?creatinine?(SCr)?and?urea?nitrogen?(BUN)?were?measured?by?sarcosine?oxidase?and?urease?method.?The?mice?were?sacrificed?under?anesthesia?at?5?days?after?operation?to?harvest?renal?tissues,?and?the?pathological?damage?of?the?kidney?was?evaluated?by?hematoxylin-eosin?(HE)?staining.?The?ultrastructure?of?mitochondria?in?mouse?renal?tubular?epithelial?cells?was?observed?under?transmission?electron?microscope.?The?levels?of?reduced? glutathione?hormone?(GSH),?malondialdehyde?(MDA)?and?nitric?oxide?synthase?(NOS)?in?mitochondrion?were?determined?by?micro-enzyme?method,?thiobarbituric?acid?method,?colorimetry?method,?respectively.?The?protein?expressions?of?Klotho,?Bcl-2?and?cytochrome?C?(Cyt?C)?were?detected?by?Western?Blot.? Results? The?pathological?structure?of?the?kidneys?in?the?Sham?group?was?clear?and?intact.?Compared?with?the?Sham?group,?the?renal?tissue?edema?of?the?mice?in?the?CLP?group?was?significant,?and?the?transparent?tube?type?was?observed?in?the?small?lumen,?and?the?interstitial?inflammatory?cells?infiltrated;?the?levels?of?SCr?and?BUN?were?significantly?increased?[SCr?(μmol/L):?182.60±6.97?vs.?47.20±5.37,?BUN?(mmol/L):?53.70±5.12?vs.?18.70±2.62,?both?P?<?0.01];?the?mitochondria?were?swollen?and?deformed,?the?sputum?structure?was?destroyed,?the?matrix?density?was?decreased,?the?outer?membrane?was?lost,?and?the?levels?of?MDA,?GSH?and?NOS?were?significantly?increased?[MDA?(μmol/g):?1.172±0.046?vs.?0.746±0.094,?GSH?(μmol/g):?5.765±0.059?vs.?4.223±0.072,?NOS?(kU/g):?0.91±0.05?vs.?0.68±0.03,?all?P?<?0.01];?the?protein?expressions?of?Klotho?and?Bcl-2??in?renal?tissue?were?decreased,?and?the?protein?expression?of?Cyt?C?was?increased?(Klotho/β-actin:?0.188±0.020?vs.?0.538±0.024,?Bcl-2/β-actin:?0.311±0.010?vs.?0.391±0.015,?Cyt?C/β-actin:?0.226±0.010?vs.?0.135±0.006,?all??P?<?0.01).?Comparing?with?the?CLP?group,?the?glomerular?and?tubular?tissue?epithelial?edema?and?the?small?lumen?in?the?CLP+KL?group?were?reduced;?the?levels?of?SCr?and?BUN?were?significantly?decreased?[SCr?(μmol/L):?85.70±7.23?vs.?182.60±6.97,?BUN?(mmol/L):?35.30±3.50?vs.?53.70±5.12,?both?P?<?0.01];?the?mitochondrial?structure?was?relatively?intact;?the?levels?of?MDA,?GSH?and?NOS?were?significantly?decreased?[MDA?(μmol/g):?0.958±0.072?vs.?1.172±0.046,?GSH?(μmol/g):?4.756±0.107?vs.?5.765±0.059,?NOS?(kU/g):?0.79±0.02?vs.?0.91±0.05,?all?P?<?0.01];?the?protein?expressions?of?Klotho,?Bcl-2?were?significantly?increased,?but?the?protein?expression?of?Cyt?C?was?significantly?decreased?(Klotho/β-actin:?0.336±0.011?vs.?0.188±0.020,?Bcl-2/β-actin:?0.474±0.017?vs.?0.311±0.010,?Cyt?C/β-actin:??0.168±0.006?vs.?0.226±0.010,?all?P?<?0.01).? Conclusion? Klotho?protein?has?significant?protective?effects?on?AKI?in?septic?mice,?and?its?mechanism?is?related?to?maintaining?mitochondrial?structural?integrity?and?oxidative?stress?response.
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Klotho (KL) protein is a membrane-bound protein mainly expressed in the kidney and brain.It can regulate the expression of multiple signal pathways and target genes,such as ion channels,transforming growth factor (TGF),and exert anti-aging effects,kidney protection,anti-tumor and regulating metabolism.Recent studies have found that KL protein plays an important role in renal diseases such as renal interstitial fibrosis (RIF).KL deficiency can induce and promote the progress of RIF,and KL overexpression or supplementation can prevent RIF.This review focuses on the anti-RIF effect of KL protein through multiple signaling pathways,and aims to provide a new direction for anti-RIF therapy.
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Objective To investigate the effect of Danhong injection on serumα-klotho protein, lipid metabolism and atherosclerotic plaque in patients with chronic renal failure undergoing hemodialysis.Methods A total of 100 patients with chronic renal failure treated in the hospital from February 2013 to April 2016 were selected and randomly divided into the control group and the observation group with 50 cases in each group.The two groups were treated with hemodialysis, and the observation group was additionally treated with Danhong injection.Changes in serumα-klotho protein, fibroblast growth factor 23 (FGF-23), lipid metabolism indexes, such as total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C), renal function indexes, such as serum creatinine (Scr), blood urea nitrogen (BUN) and endogenous creatinine clearance rate (Ccr) in the two groups were determined before and after treatment.Carotid artery ultrasonography was performed to observe changes in the size, number and thickness of atherosclerotic plaques and carotid intima-media thickness (IMT) before and after treatment.Results FGF-23, TC, TG, Scr and BUN in the observation group were decreased after 8 weeks of treatment[ (1 526.25±46.87) pg/mL, (5.21±1.23) mmol/L, (1.26±0.23) mmol/L, (390.11±52.26) μmol/L, (14.11±3.26) mmol/L vs. (1 768.74±54.26) pg/mL, (5.93±0.26) mmol/L, (1.63±0.14) mmol/L, (443.26±47.85) μmol/L, (18.52±2.79) mmol/L], whileα-Klotho protein, HDL-C and Ccr were increased[ (790.44±40.61) pg/mL, (1.21±0.21) mmol/L, (37.41±3.26) mL/min vs. (662.25±76.54) pg/mL, (1.00±0.29) mmol/L, (33.51±3.41) mL/min].Compared with those in the control group, there were statistically significant differences (t=23.914, 4.049, 9.716, 5.304, 7.267, 10.461, 4.147, 5.845, P＜0.05).In 6 months of follow-up, plaques shrank, the thickness of plaque and IMT decreased, and the number of plaques decreased in the observation group[ (0.06±0.01) cm2, (1.11±0.23) mm, (1.01±0.23) mm, (2.86±0.51) vs. (0.10±0.06) cm2, (1.87±0.49) mm, (1.43±0.20) mm, (3.41±1.03) ].Compared with those in the control group, the differences were statistically significant (t=4.649, 9.928, 9.743, 3.383, all P＜0.05).Conclusion Danhong injection can increase the expression level ofα-klotho protein in serum of patients with chronic renal failure undergoing hemodialysis, down regulate the expression of FGF-23, improve lipid metabolism and renal function in patients, reduce the degree of atherosclerosis, the number of patches and the thickness of plaque.
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Objective To evaluate the clinical significance of serum Klotho protein levels in the early diagnosis and prognosis of acute kidney injury (AKI) among adult patients in the intensive care units (ICU).Methods The study was prospective and observational.Blood samples and clinical data of AKI patients admitted to the ICU of the First Affiliated Hospital of Xinjiang Medical University between July 1 and August 31,2016 were collected.ELISA was used for the detection of Klotho and NGAL.Receiver operating characteristic curve (ROC) and the area under the curve (AUC) were used to compare the predictive performance among Klotho,NGAL and serum creatinine,evaluating the sensitivity and specificity of Klotho on the diagnosis of AKI.The correlation between Klotho and prognosis of AKI was investigated by comparing serum Klotho levels and early AKI predictors.Results The patients were divided into AKI group of 52 cases and non-AKI group of 98 cases.The baseline serum Klotho level in AKI group was significantly lower than that in non-AKI group (P ＜ 0.001).The AUC of Klotho predicting for AKI was 0.945(95％ CI:0.892-0.997) and the best cut off value was 1.76 μg/L(sensitivity 92％,specificity 94％).The predictive ability of Klotho was significantly higher than serum creatinine (Scr),and the sensitivity is higher than NGAL (sensitivity 87％,specificity 96％).Serum Klotho combined with Scr predicted better AKI (AUC=0.958,95％ CI:0.915-1.000,sensitivity 96％,sensitivity 92％).The level of Klotho in patients with AKI was significantly different between the renal function recovery group and non-recovery group (P=0.047),while there was no significant difference between the two groups in the level of NGAL and Scr (P ＞ 0.05).There was no significant correlation between the Klotho level at diagnosis of AKI and peak Scr,peak eGFR,Scr at discharge and eGFR at discharge (r=0.026,P=0.853;r=-0.127,P=0.368;r=0.243,P=0.082;r=-0.187,P=0.184).Conclusion Serum Klotho may be a potential biomarker for early diagnosis of AKI,but the association between serum klotho and the prognosis of AKI requires further study.
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Objective To investigate the relationship between FGF23 and Klotho in GD patients. Methods From March 2016 to November 2016, forty-three newly diagnosed and untreated Grave disease patients (GD group) and 27 healthy subjects were selected. Meanwhile,Peripheral venous serum was collected to detect serum calcium, phosphorus and thyroid function in GD group. The levels of FGF23, Klotho protein and 1 in each group were detected by enzyme-linked immunosorbent assay ( ELISA ) . The expression levels of FGF23,Klotho protein and 1,25-dihydroxyvitamin D3 were detected by enzyme-linked immunosorbent assay ( ELISA ) . The differences of FGF23, Klotho protein, 1, 25-dihydroxyvitamin D3, serum calcium and phosphorus between GD group and normal control group were compared,and the correlation between each index was analyzed. The relationship between FGF23 and Klotho protein and the pathogenesis of GD was explored. Results ( 1) Thyrotropin in thyroid function was significantly lower in GD group than that in normal group,and the difference was statistically significant ( 0. 003 ( 0. 002, 0. 004 ) mU/L vs. 1. 650 ( 0. 81, 2. 14 ) mU/L, Z=- 7. 587,P<0. 05] . Thyrotropin receptor antibodies,free triiodothyronine and free thyroxine in GD group were significantly higher than those in normal group. There were [ thyroid stimulating hormone receptor antibodies:9. 03(3. 89,21. 29)) U/L vs. 0. 60(0. 38,0. 97)) U/L,free triiodothyronine:23. 36(11. 61,38. 00)) pmol/L vs. 4. 63(4. 03,4. 92)) pmol/L,free thyroxine:4. 34(33. 94,100. 00) pmol/Lvs. 17. 69(15. 80,20. 35) pmol/L;Z=-6. 694,-6. 878,-6. 836,P<0. 05];( 2) The serum levels of FGF23,Klotho and phosphorus in patients with hyperthyroidism were significantly higher than those in the normal group,and the difference was statistically significant [FGF23:(524. 2±66. 7) ng/L vs. (467. 2±64. 5) ng/L,Klotho:8. 29(6. 89,11. 37) pg/ml vs. 6. 69 (6. 36,7. 53) pg/ml,phosphorus:1. 33(1. 03,1. 52) mmol/L vs. 1. 02(0. 84,1. 20) mmol/L; t=3. 517,Z=-3. 936,-3. 795,P<0. 05]. The Results of correlation analysis showed that: (1) There was no correlation among FGF23 and Klotho,thyroid stimulating hormone receptor antibody,free thyroxine,1,25-dihydroxyvitamin D3,phosphorus and calcium a ( P>0. 05);( 2) There was no correlation among Klotho and thyroid stimulating hormone receptor antibody, free thyroxin, 1, 25-dihydroxyvitamin D3, phosphorus and calcium ( P>0. 05 ) . Conclusion The elevated expression of FGF23 in GD may be involved in the pathogenesis of GD, and the elevated expression of Klotho in GD may be due to the abnormal immune status in GD patients,which may play a protective role.
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OBJECTIVE@#To observe the effect of Ronghuang granule on serum fibroblast growth factor 23 (FGF23), fibroblast growth factor receptor (FGFRs) and Klotho protein levels in non-dialysis patients with chronic kidney disease-mineral and bone disorder (CKD-MBD) and kidney deficiency and damp heat syndrome.@*METHODS@#Seventy non-dialysis CKD-MBD patients with kidney deficiency and dampness-heat syndrome were randomized into control group (=35) and treatment group (=35). All the patients were given routine treatment combined with traditional Chinese medicine retention enema, and the patients in the treatment group received additional Ronghuang granule treatment (3 times a day). After the 12-week treatments, the patients were examined for changes of TCM syndromes. Serum levels of Ca, P, parathyroid hormone (iPTH), FGF23, FGFRs and Klotho proteins were detected before and after treatment. These parameters were also examined in 20 healthy volunteers.@*RESULTS@#Sixty-five patients completed the study, including 33 in the control group and 32 in the treatment group. The patients in the treatment group showed significantly better treatment responses than those in the control group ( < 0.05 or 0.01). At 4, 8, and 12 weeks of treatment, the patients in the treatment group had significantly lowered scores of TCM syndromes compared with the score before treatment ( < 0.05 or 0.01), while in the control group, significant reduction of the scores occurred only at 12 weeks ( < 0.05); at each of the time points, the treatment group had significantly greater reductions in the score than the control group ( < 0.01). Significant improvements in serum Ca, P and iPTH levels were observed at 4, 8, and 12 weeks in the treatment group ( < 0.05) but only at 12 weeks in the control group ( < 0.05). The patients in the control and treatment groups all showed elevated serum levels of FGF23, FGFRs and Klotho protein compared with the normal subjects ( < 0.01); FGF23, FGFRs and Klotho levels were significantly reduced in the treatment group ( < 0.05) but remained unchanged in the control group (>0.05), showing significant differences between the two groups.@*CONCLUSIONS@#Ronghuang granule improves the clinical symptoms of non-dialysis CKD-MBD patients with kidney deficiency and dampness heat syndrome by reducing serum levels of FGF23, FGFRs and Klotho, improving calcium and phosphorus metabolism disorder, and inhibiting secondary hyperparathyroidism.
Subject(s)
Humans , Calcium , Blood , Chronic Kidney Disease-Mineral and Bone Disorder , Blood , Therapeutics , Drugs, Chinese Herbal , Pharmacology , Enema , Fibroblast Growth Factors , Blood , Glucuronidase , Blood , Parathyroid Hormone , Blood , Phosphorus , Blood , Receptors, Fibroblast Growth Factor , Blood , Renal Insufficiency, Chronic , Blood , Therapeutics , Sweating Sickness , Blood , Therapeutics , SyndromeABSTRACT
Objective To investigate the effect of klotho on the human vein umbilical endothelial cells (HUVECs) injury induced by indoxyl sulfate (IS) and to explore its mechanism and the role of endoplasmic reticulum stress (ERS) in this process.Methods (1) The cell vitalities of HUVECs incubated with different concentration of IS (5,25,50 mg/L) for 48 h and with 50 mg/L IS fordifferent time points (12,24,48 h) were measured by CCK-8 assay.(2) HUVECs were incubated with 50 mg/L IS and different concentration of klotho (0,1,10,100 μg/L) for 48 h and their cell viabilities were measured by CCK-8 assay.(3) HUVECs were divided into four groups:control group,IS group (50mg/L IS),klotho group (50 mg/L IS+ 100 μg/L klotho) and Compound C group (50 mg/L IS+100 μg/L klotho+ 10 μmol/L Compound C).The cell vitality and the apoptosis of HUVECs were evaluated by CCK-8 assay and flow cytometry,respectively.The mRNA and protein expressions of GRP78 and CHOP were measured by real-time PCR and Western blotting.The phosphorylation level of AMPK was tested by Western blotting.Results IS inhibited cell vitality in the time-dependent and concentration-dependent manner.The cell viability of HUVECs with 50 mg/L IS was lower than normal control (P＜0.05).The inhibited cell vitality induced by IS was partly restored by klotho in concentration-dependent manner.The cell viability was higher in 100 μg/L klotho+50 mg/L IS group than 50 mg/L IS group (P ＜ 0.05).Compared with control group,the expressions of GRP78 and CHOP and cell apoptosis increased,however,the level of phosphorylated AMPK (p-AMPK) decreased in IS group (all P ＜ 0.05).Compared with IS group,the expressions of GRP78 and CHOP and cell apoptosis decreased and the level of p-AMPK increased in klotho group (all P ＜ 0.05).Furthermore,the above effects of klotho could be partly blocked by Compound C.The above indexes showed statistical differences between Compound C group and klotho group.Conclusions IS can inhibit the HUVECs cell vitality,and induce ERS and cell apoptosis.Klotho protein could antagonize the above effects,probably through activating AMPK pathway and reducing ERS-mediated cell apoptosis.
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Objective To investigate the effect of klotho on the human vein umbilical endothelial cells (HUVECs) injury induced by indoxyl sulfate (IS) and to explore its mechanism and the role of endoplasmic reticulum stress (ERS) in this process.Methods (1) The cell vitalities of HUVECs incubated with different concentration of IS (5,25,50 mg/L) for 48 h and with 50 mg/L IS fordifferent time points (12,24,48 h) were measured by CCK-8 assay.(2) HUVECs were incubated with 50 mg/L IS and different concentration of klotho (0,1,10,100 μg/L) for 48 h and their cell viabilities were measured by CCK-8 assay.(3) HUVECs were divided into four groups:control group,IS group (50mg/L IS),klotho group (50 mg/L IS+ 100 μg/L klotho) and Compound C group (50 mg/L IS+100 μg/L klotho+ 10 μmol/L Compound C).The cell vitality and the apoptosis of HUVECs were evaluated by CCK-8 assay and flow cytometry,respectively.The mRNA and protein expressions of GRP78 and CHOP were measured by real-time PCR and Western blotting.The phosphorylation level of AMPK was tested by Western blotting.Results IS inhibited cell vitality in the time-dependent and concentration-dependent manner.The cell viability of HUVECs with 50 mg/L IS was lower than normal control (P＜0.05).The inhibited cell vitality induced by IS was partly restored by klotho in concentration-dependent manner.The cell viability was higher in 100 μg/L klotho+50 mg/L IS group than 50 mg/L IS group (P ＜ 0.05).Compared with control group,the expressions of GRP78 and CHOP and cell apoptosis increased,however,the level of phosphorylated AMPK (p-AMPK) decreased in IS group (all P ＜ 0.05).Compared with IS group,the expressions of GRP78 and CHOP and cell apoptosis decreased and the level of p-AMPK increased in klotho group (all P ＜ 0.05).Furthermore,the above effects of klotho could be partly blocked by Compound C.The above indexes showed statistical differences between Compound C group and klotho group.Conclusions IS can inhibit the HUVECs cell vitality,and induce ERS and cell apoptosis.Klotho protein could antagonize the above effects,probably through activating AMPK pathway and reducing ERS-mediated cell apoptosis.
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Objective To explore the role of serum levels of Klotho protein and fibroblast growth factor(FGF)-23 in patients with chronic kidney disease(CKD).Methods A total of 160 patients with CKD were recruited into CKD group in this study,160 healthy controls were selected in the control group from March 2014 to March 2016.Basic clinical data,blood biochemical index,serum level of Klotho protein and FGF-23 were analyzed and compared between the two groups.Results The hemoglobin,serum albumin,blood calcium,glomerular filtration rate,Klotho protein levels in the CKD group were lower than those in the control group,and the serum creatinine,blood urea,blood serum phosphorus and FGF-23 were higher than those in the control group(P<0.05).With the progress of CKD stages,hemoglobin,glomerular filtration rate,Klotho protein levels gradually decreased,serum creatinine,blood urea,blood serum phosphorus,FGF-23 level increased(P<0.05).The levels of FGF-23 were negatively related to hemoglobin,glomerular filtration rate,Klotho(r=-0.584,0.692,-0.724)and were positively correlated to serum creatinine,blood urea,blood serum phosphorus(r=0.814,0.703,0.527).The levels of Klotho protein were positively related to hemoglobin,glomerular filtration rate(r=0.612,0.685),and were negatively correlated to serum creatinine,blood serum phosphorus,blood urea,FGF-23(r=-0.720,-0.690,-0.519,0.724).Conclusion High concentrations of FGF-23 and Klotho protein with low concentration were not only related to calcium phosphate metabolic disorders of patients with CKD,and were also associated with the prognosis of patients with CKD,which might be early biomarkers and predictor in patients with CKD.
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AIM:To study the protective effect of anti-aging Klotho protein on human umbilical vein endothe-lial cells (HUVECs) treated with high glucose (HG).METHODS:HUVECs were cultured in vitro, and divided into PBS control group, 5.5 mmol/L glucose group, 33.3 mmol/L glucose group, 0.1 μmol/L Klotho +33.3 mmol/L glucose group, 1 μmol/L Klotho+33.3 mmol/L glucose group , and 10μmol/L Klotho+33.3 mmol/L glucose group .The viabili-ty of the HUVECs was measured by MTT assay .The content of malondialdehyde ( MDA) , and the activities of lactate de-hydrogenase (LDH), superoxide dismutase (SOD) and glutathione (GSH) in cell culture supernatants were observed . The production of reactive oxygen species ( ROS) in HUVECs was analyzed by flow cytometry .The levels of nitric oxide ( NO) , endothelin ( ET-1 ) , intercellular adhesion molecule-1 ( ICAM-1 ) in HUVEC culture medium were detected by ELISA.The protein expression of nuclear factor-kappa B (NF-κB) in the HUVECs was determined by Western blot .RE-SULTS:Compared with PBS control group , 33.3 mmol/L glucose significantly decreased the HUVEC viability , increased ROS, LDH and MDA levels , reduced the activities of SOD and GSH , decreased the NO secretion , and induced the ET-1 and ICAM-1 secretion and the protein expression of NF-κB in HUVECs.When HUVECs were treated with Klotho protein at different concentrations combined with 33.3 mmol/L glucose, the cell viability was increased significantly , the ROS, LDH and MDA levels were decreased significantly , the antioxidant SOD and GSH activities were significantly increased , the se-cretion of NO was increased , but ET-1 and ICAM-1 releases and protein expression of NF-κB were significantly reduced . CONCLUSION:Anti-aging Klotho protein promotes the viability of HUVECs treated with HG , reduces the oxidative dam-age and ROS production , and restores the normal secretory function of HUVECs , thus playing a protective role in vascular endothelial cells through reducing the protein expression of NF-κB.
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Objective To discuss the effects of multimodal combination dialysis on Klotho protein, fibroblast growth factor-23 (FGF-23) and brain natriuretic peptide (BNP) in patients with maintenance hemodialysis (MHD). Methods A randomized controlled trial (RCT) was conducted. 120 patients who was diagnosed with chronic renal failure (CRF) uremia receiving MHD over 3 months admitted to Blood Purification Centre of Department of Nephrology of the Second People's Hospital of Guiyang from December 2015 to December 2016 were enrolled, who were randomly divided into hemodialysis (HD) group (HD for 8 times a month), HD + hemofiltration (HF) group (HD for 8 times a month + HF once a month), and HD + HF + hemoperfusion (HP) group (HD for 8 times a month + HF for 4 times a month + HP once a month), with 40 patients in each group. Before and after treatment for 6 months and 12 months, blood was taken from venous circuit tube, the serum Klotho protein and FGF-23 levels were determined by enzyme linked immunosorbent assay (ELISA), and the serum BNP level was determined by electrochemiluminescence. Results 120 patients with MHD were enrolled in the final analysis without withdrawal. There were no significant differences in the levels of Klotho protein, FGF-23, or BNP before enrollment among the three groups (all P > 0.05). Compared with those before enrollment, the levels of serum Klotho protein after enrollment in three groups showed a sustained upward tendency, which were higher in HD + HF + HP group than in HD + HF group and HD group (μg/L: 2.59±0.61, 1.63±0.35, 1.13±0.26 at 6 months, F = 119.374, P = 0.000; 6.98±1.21, 3.57±1.03, 2.12±0.43 at 12 months, F = 275.675, P = 0.000); the levels of FGF-23 showed a sustained downward tendency, which were lower in HD + HF + HP group than in HD + HF group and HD group (ng/L: 69.22±38.26, 132.28±61.18, 178.50±74.64 at 6 months, F = 33.509, P = 0.000; 32.81±17.32, 87.93±43.27, 146.33±69.28 at 12 months, F = 55.466, P = 0.000);the BNP showed a similar tendency as FGF-23 (ng/L: 4083.39±2864.53, 7245.69±4643.81, 7969.12±5360.85 at 6 months, F = 8.758, P = 0.000; 1521.86±894.63, 4554.32±1969.84, 5013.89±2033.64 at 12 months, F = 49.003, P = 0.000). Conclusion Multimodal combination dialysis can increase the Klotho protein level, and decrease the levels of FGF-23 and BNP in MHD patients with CRF uremia.
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Objective:To analyze the levels of serum calcium,phosphate,fibroblast growth factor 23 (FGF23),and Klotho proteins in patients with chronic kidney disease (CKD),and to investigate the correlations of FGF23 and Klotho proteins with cardiac complicates in patients with chronic kidney disease-mineral and bone disorder (CKD-MBD).Methods:A total of 180 CKD-MBD patients were enrolled for this study.Among them,60 patients underwent regular hemodialysis,60 patients did not undergo renal replacement therapy and 60 patients were diagnosed as second hyperparathyroidism (SHPT).Thirty age and gendermatched health volunteers served as controls.Serum samples were collected and tested,and the demographical,clinical and biochemical data were all recorded.FGF23 and Klotho levels in serum samples were analyzed by enzyme-linked immunosorbent assay.Data of echocardiography and plain abdominal X rays were collected as well.The influential factors for cardiovascular injury,the relationship between biochemical indexes and ectopic calcification,and the correlations of FGF23 and Klotho with cardiac complicates were analyzedResults:Patients,who kept hemodialysis,especially those with SHPT,exhibited an increase in serum FGF23 level while a decrease in serum Klotho protein levels (P＜0.01).Patients with higher levels of serum FGF23 were more likely to have ectopic calcification (OR=4.667),while patients with lower levels of serum Klotho had high risks to get myocardial hypertrophy (OR=3.496).Receiver operator characteristic (ROC) curve analysis showed that the area under the curve (AUC) for FGF23 was 0.778 (P＜0.01) while for Klotho was 0.715 (P＜0.01).Conclusion:Patients,who kept hemodialysis,especially those with SHPT,have a significant increase in serum FGF23 protein levels and a significant decrease in serum Klotho protein levels.Serum FGF23 and Klotho protein levels are closely correlated with left ventricular enlargement and hypertrophy.Serum FGF23 and Klotho protein are risk factors for heart.