Correlation of serum soluble Klotho protein and tendino-C with severity of disease and oxidative stress in children with IgA nephropathy / 国际检验医学杂志

Objective To analyze the correlation between serum soluble Klotho protein(sKL)and tendino-C(TN-C)and the severity of disease and oxidative stress in children with immunoglobulin A(IgA)nephropa-thy.Methods A total of 85 children with IgA nephropathy admitted to the hospital from July 2019 to August 2022 were selected as IgA nephropathy group,and 85 healthy patients who underwent physical examination in the hospital during the same period were selected as healthy group.Serum sKL and TN-C levels were com-pared between the two groups.Receiver operating characteristic(ROC)curve was drawn to analyze the value of serum sKL,TN-C and their combination in predicting the occurrence of IgA nephropathy.IgA nephropathy group was divided into mild group(28 cases),moderate group(39 cases)and severe group(18 cases)accord-ing to 24 h urinary protein quantity.Serum sKL,TN-C levels and oxidative stress indexes[malondialdehyde(MDA),superoxide dismutase(SOD)and advanced oxidation protein product(AOPP)]of the three groups were compared.The correlation between serum sKL and TN-C levels and oxidative stress indexes was ana-lyzed by Spearman correlation,and the correlation between serum sKL and TN-C levels and the severity of IgA nephropathy in children was examined by Kendall's Tau-b.Results The serum sKL level in IgA ne-phropathy group was lower than that in healthy group,and the serum TN-C level was higher than that in healthy group,the difference was statistically significant(P＜0.05).ROC curve showed that the area under the curve and 95％CI of serum sKL,TNC and their combination predicted the occurrence of IgA nephropathy were 0.726(95％CI:0.648-0.803),0.853(95％CI:0.796-0.909)and 0.891(95％CI:0.845-0.937).The level of serum sKL in severe group was lower than that in moderate and mild groups,while the level of serum TN-C was higher than that in moderate and mild groups,and the difference was statistically significant(P＜0.05).The serum SOD level of severe group was lower than that of moderate group and mild group,and the serum AOPP and MDA levels were higher than those of moderate group and mild group,the difference was statistically significant(P＜0.05).Spearman correlation showed that sKL level was positively correlated with SOD(r＞0,P＜0.05),and negatively correlated with AOPP and MDA(r＜0,P＜0.05).TN-C level was negatively correlated with SOD(r＜0,P＜0.05),and positively correlated with AOPP and MDA(r＞0,P＜0.05).Conclusion The levels of serum sKL and TN-C in children with IgA nephropathy are related to the severity of disease and oxidative stress,and the combination of SKL and TN-C can effectively predict the occurrence of IgA nephropathy.

The correlation between serum Klotho levels and frailty in elderly people / 中华老年医学杂志

Objective:To examine the correlation between serum Klotho levels and frailty in elderly people.Methods:In this cross-sectional study, 150 community-dwelling elderly people aged 65 years and over were enrolled.Subjects were divided into a frail(n=50, 33.3%), a pre-frail(n=47, 31.3%)and a non-frail(n=53, 35.3%)group based on the Fried phenotype.General participant data, routine laboratory test results, short physical performance battery(SPPB)results and human body composition data were collected.Serum Klotho protein levels were measured by an enzyme-linked immunosorbent assay.The relationship between serum Klotho protein levels and frailty was analyzed by using Spearmen's correlation analysis and Logistic regression analysis.Results:Klotho protein levels were lower in the frail group than in the non-frail group( P=0.001), whereas differences between the frail group and the pre-frail group and between the pre-frail group and the non-frail group were not statistically significant(all P>0.05).When Klotho protein levels were classified into four quartiles, i.e., Q 1, Q 2, Q 3, and Q 4, using three cut-off vales(2.28, 3.52, and 5.09 mg/L), the prevalences of frailty were 51.4%(19/37), 39.5%(15/38), 24.3%(9/37)and 18.4%(7/38), respectively.The prevalence of frailty decreased with increasing Klotho protein levels( χ2=11.204, P=0.011).Spearman correlation analysis showed that the Klotho protein level was negatively correlated with frailty( r=-0.310, P<0.001).Multivariate Logistic regression analysis results showed that age( OR=1.109, 95% CI: 1.011-1.217, P=0.028)and sarcopenia( OR=6.511, 95% CI: 1.279-33.147, P=0.024)were risk factors for frailty, while walking( OR=0.104, 95% CI: 0.033-0.326, P<0.001), a high SPPB score( OR=0.780, 95% CI: 0.627-0.970, P=0.026), and a high Klotho protein level( OR=0.752, 95% CI: 0.581-0.974, P=0.031)were protective factors against frailty. Conclusions:The serum Klotho protein level may be used as a parameter for the assessment of frailty.It is negatively correlated with frailty, suggesting that elderly people with low serum Klotho protein levels are at high risk of developing frailty.

Traditional Chinese Medicine Intervenes in Signaling Pathways Associated with CKD-MBD： A Review / 中国实验方剂学杂志

Traditional Chinese medicine （TCM） has certain advantages in the treatment of chronic kidney disease-mineral and bone disorder （CKD-MBD）. In recent years， there have been many studies on the treatment of CKD-MBD by Chinese medicinal compounds and monomers. As revealed by literature retrieval， the research on the mechanism of Chinese medicine in intervening in signaling pathways related to CKD-MBD was mainly based on self-made Chinese medicinal compounds， and the action pathways involved fibroblast growth factor 23/Klotho （FGF23/Klotho） signaling pathway， Wnt/β-catenin signaling pathway， receptor activator of nuclear factor-κB/receptor activator of nuclear factor-κB ligand/osteoprotegerin （RANK/RANKL/OPG） system， and other signaling pathways. TCM can improve calcium and phosphorus metabolism and bone metabolism disorder， and regulate inflammatory reaction， oxidative stress， apoptosis， and autophagy by regulating this series of signaling pathways for the treatment of CKD-MBD. This paper introduced the research results of these signaling pathways and the mechanism of TCM in the treatment of CKD-MBD in order to provide ideas and references for the related research of Chinese medicine in the treatment of CKD-MBD.

Relationship between serum MLT and Klotho protein and pro-inflammatory factors and efficacy in patients with primary angle-closure glaucoma / 国际检验医学杂志

Objective To investigate the relationship of serum melatonin(MLT)and Klotho protein with pro-inflammatory factors and the effect of surgical treatment in patients with primary angle-closure glaucoma(PACG).Methods A total of 149 PACG patients admitted to the Zigong Fourth People's Hospital from June 2018 to June 2021 were selected as the case group,and 149 healthy people who underwent physical examina-tion in the hospital during the same period were selected as the control group.The levels of MLT,Klotho pro-tein,interleukin-6(IL-6),interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)were compared be-tween the two groups.Pearson correlation analysis was used to analyze the correlation between MLT,Klotho protein and pro-inflammatory factors.Univariate and multivariate Logistic regression models were used to an-alyze the influencing factors of surgical treatment.Results Compared with the control group,the levels of MLT and Klotho protein in the case group were decreased(P<0.05),and the levels of IL-1β,IL-6 and TNF-α were increased(P<0.05).Pearson correlation analysis showed that serum MLT and Klotho protein were negatively correlated with the levels of pro-inflammatory factors IL-1β,IL-6,and TNF-α(P<0.05).After 12 months of surgical treatment,30 patients(20.13%)had no effect of surgical treatment(ineffective group).119 patients(79.87%)had effective surgical treatment(effective group).The univariate analysis showed that compared with the effective group,the preoperative intraocular pressure(IOP),proportion of chronic PACG,preoperative best corrected visual acuity,proportion of preoperative Angle adhesion>160° were significantly increased(P<0.05),and the preoperative anterior chamber depth,MLT,and Klotho protein levels were de-creased in the ineffective group(P<0.05).Multivariate Logistic regression analysis showed that elevated pre-operative IOP,chronic PACG,decreased MLT and Klotho levels were independent risk factors for ineffective surgical treatment(P<0.05).Conclusion serum MLT,Klotho protein levels in patients with PACG,closely associated with proinflammatory factor,is the related factors influencing the effect of surgical treatment.

Klotho and FGF23 in Small for gestational age infants update / 中国综合临床

Klotho protein is an anti-aging gene product, which is involved in the regulation of calcium, phosphate and vitamin D metabolism together with fibroblast growth factor 23 (FGF23). In recent years, studies have found that Klotho/FGF23 is closely related to the growth and development of children and newborns. This paper summarizes the role of Klotho/FGF23 in the occurrence and development of infants small for gestational age, in order to provide further understanding and inspiration for the prevention and treatment of SGA complications.

Association of Klotho with cerebrovascular disease and associated biological mechanisms / 中风与神经疾病杂志

@#Cerebrovascular disease is one of the leading causes of death and disability across the world. Vascular homeostasis and recovery is critical to the body. Klotho is an important endogenous pleiotropic protein involved in a variety of pathophysiological processes,such as aging and calcium and phosphorus metabolism,which is closely associated with vascular disease. Recent clinical studies have demonstrated close links between low Klotho levels and various risk factors for cerebrovascular disease,suggesting that Klotho has the potential of predicting the risk of ischemic and hemorrhagic cerebrovascular diseases and poor prognosis. Basic studies have also shown that Klotho plays a crucial role in the maintenance of vascular homeostasis. Klotho can promote the production of nitric oxide,inhibit the expression of inflammatory factors,and mediate antioxidant and anti-aging effects,which can delay atherosclerosis and vascular calcification. Therefore,Klotho may serve as a new target for the prevention and treatment of cerebrovascular disease. This review provides an overview of the relationship between Klotho and cerebrovascular disease and its potential biological mechanisms.

Research Progress of β-Klotho in Energy Metabolism and Tumor Development / 中国生物化学与分子生物学报

β-Klotho (KLB) is a member of the Klotho protein family, which is mainly distributed in organs and tissues such as the liver, fat, pancreas, and brain. KLB is a single-pass transmembrane protein whose structural characteristics determine that KLB acts as a co-receptor for fibroblast growth factor (FGF) 19/21 targeting the activation of fibroblast growth factor receptor (FGFRs). KLB is involved in the regulation of blood glucose, lipids, body weight, bile acid circulation, and hepatocyte proliferation in the FGF21/19-KLB-FGFRs pathway. This paperwill review the structural characteristics and distribution of KLB, as well as the regulatory mechanism of material energy and its role in tumor formation in the FGF19/21-KLB-FGFRs pathways.

Effect of recombinant human growth hormone on serum Klotho and fibroblast growth factor 23 in children with idiopathic short stature / 中国当代儿科杂志

OBJECTIVES@#To investigate the changes in the serum levels of Klotho, fibroblast growth factor 23 (FGF23), and insulin-like growth factor-1 (IGF-1) in children with idiopathic short stature (ISS) before and after recombinant human growth hormone (rhGH) treatment, as well as the correlation of Klotho and FGF23 with the growth hormone (GH)/IGF-1 growth axis in these children.@*METHODS@#A prospective study was conducted on 33 children who were diagnosed with ISS in the Department of Pediatrics, Hebei Provincial People's Hospital, from March 10, 2021 to December 1, 2022 (ISS group). Twenty-nine healthy children, matched for age and sex, who attended the Department of Child Healthcare during the same period, were enrolled as the healthy control group. The children in the ISS group were treated with rhGH, and the serum levels of Klotho, FGF23, and IGF-1 were measured before treatment and after 3, 6, and 9 months of treatment. A correlation analysis was conducted on these indexes.@*RESULTS@#There were no significant differences in the serum levels of IGF-1, Klotho, and FGF23 between the ISS and healthy control groups (P>0.05). The serum levels of Klotho, FGF23, and IGF-1 increased significantly in the ISS group after 3, 6, and 9 months of rhGH treatment (P<0.05). In the ISS group, Klotho and FGF23 levels were positively correlated with the phosphate level before treatment (P<0.05). Before treatment and after 3, 6, and 9 months of rhGH treatment, the Klotho level was positively correlated with the IGF-1 level (P<0.05), the FGF23 level was positively correlated with the IGF-1 level (P<0.05), and the Klotho level was positively correlated with the FGF23 level (P<0.05), while Klotho and FGF23 levels were not correlated with the height standard deviation of point (P>0.05).@*CONCLUSIONS@#The rhGH treatment can upregulate the levels of Klotho, FGF23, and IGF-1 and realize the catch-up growth in children with ISS. Klotho and FGF23 may not directly promote the linear growth of children with ISS, but may have indirect effects through the pathways such as IGF-1 and phosphate metabolism. The consistent changes in Klotho, FGF23 and IGF-1 levels show that there is a synergistic relationship among them in regulating the linear growth of ISS children.

Research Progress of Klotho in Lung Neoplasms / 中国肺癌杂志

Klotho gene was originally discovered as an anti-aging gene, Klotho protein encoded by Klotho gene is expressed in multiple human tissues, and its most prominent function is the regulation of phosphate homeostasis. Klotho protein possesses various activities, including inhibition of multiple signaling pathways, reducing oxidative stress and suppressing inflammation, and these activities are associated with cancer. Klotho protein is discovered as a universal tumor suppressor, and its expression is associated with tumorigenesis and prognosis of patients. Lung cancer is the most common malignancy tumor, and it is the leading cause of cancer deaths worldwide because of its high incidence and mortality. This article summarizes the research progress of the role of Klotho on pathogenesis, therapeutic effect and prognosis in lung cancer, in order to provide new biomarker and target for diagnosis, treatment and prognosis of lung cancer.
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Significance of serum FGF19, Klotho and FGFR4 protein expression in patients with primary biliary cirrhosis / 中国基层医药

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.

Klotho promotes high-glucose-mediated autophagy of human renal tubule cells / 中国医师杂志

Objective:To investigate the effects of Klotho on autophagy of human renal tubule cells under high glucose through AMP-activated protein kinase (AMPK) and extracellular signal regulated kinase (ERK) pathways.Methods:Human renal tubular epithelial cells cultured in vitro were divided into control group and high glucose group (HG group, added with 30 mmol/L glucose); According to the transfection of pcDNA3.1-vector or pcDNA3.1-Klotho, they were divided into two groups: Vector group and Klotho group; According to whether AMPK inhibitor compound C or ERK inhibitor curcumin was added after pcDNA3.1-Klotho transfection and high glucose stimulation, they were divided into four groups: HG+ Vector group, HG+ Klotho group, HG+ Klotho+ compound C group and HG+ Klotho+ curcumin group. The expression of Klotho was detected by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) and Western blot; The relative expression of LC3-Ⅱ/LC3-Ⅰ and p-AMPK/AMPK and p-ERK/ERK were detected by Western blot; Changes of autophagosome in human renal tubular epithelial cells observed by transmission electron microscope. Results:The protein and mRNA expression of Klotho in human renal tubular epithelial cells of HG group was significantly lower than that in the control group (all P<0.05); The LC3-Ⅱ/LC3-Ⅰ in Klotho group was significantly higher than that in Vector group ( P<0.05); The number of autophagosomes in Klotho group was also significantly higher than that in Vector group ( P<0.05); p-AMPK/AMPK in Klotho group was significantly higher than that in Vector group ( P<0.05), while p-ERK/ERK in Klotho group was significantly lower than that in Vector group ( P<0.05). The protein relative expression of p-AMPK/AMPK in HG+ Klotho+ compound C group (0.44±0.04) was significantly lower than that in HG+ Klotho group (0.79±0.08) ( P<0.01); The protein relative expression of p-ERK/ERK in HG+ Klotho+ curcumin group (1.05±0.12) was significantly higher than that in HG+ Klotho group (0.56±0.05) ( P<0.01). The relative expression of LC3-Ⅱ/LC3-Ⅰ protein in HG+ Klotho+ compound C group and HG+ Klotho+ curcumin group (0.79±0.12; 0.68±0.09) were significantly lower than that in HG+ Klotho group (1.65±0.20) (all P<0.01). Conclusions:Klotho can enhance autophagy of human renal tubular epithelial cells under high glucose condition by activating AMPK and inhibiting ERK pathway.

Research progress on the relationship between Klotho and the growth of children and newborns / 中国综合临床

Klotho is a gene associated with aging, the transmembrane protein encoded by this gene is highly expressed in the kidney, and is also expressed in tissues such as the brain, parathyroid and pituitary glands. The extracellular domain of Klotho can also be cleaved and shed to form soluble Klotho, which acts as a circulating hormone and can be detected in blood, cerebrospinal fluid, and urine. More and more studies have shown that Klotho protein plays an important role in the complex regulation of growth hormone (GH)-insulin-like growth factor 1(IGF1) axis. The interaction between Klotho protein and GH-IGF1 axis is bidirectional, which regulates each other, and then regulates the normal linear growth of children. In addition, Klotho protein can also affect the growth and development of fetus and newborn through different ways, and its mechanism is not very clear.

Hyperbaric oxygen therapy can augment the effectiveness of cognition training for persons with vascular cognitive impairment / 中华物理医学与康复杂志

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.

Mechanism of Modified Liuwei Dihuangtang in Bone Protection of CKD-MBD Model Rats： An Exploration Based on Klotho-FGF23 Axis / 中国实验方剂学杂志

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 D₃［1，25（OH）₂D₃］ 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^-1·d^-1），the ones in the modified Liuwei Dihuangtang group with modified Liuwei Dihuangtang （2.556 g·kg^-1·d^-1） ， and those in the calcitriol group with calcitriol （0.09 μg·kg^-1·d^-1）. 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）₂D₃ 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）₂D₃， 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.

Klotho protein and chronic kidney disease in children / 中华实用儿科临床杂志

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.

Z-Guggulsterone alleviated renal fibrosis and G2/M cycle arrest through Klotho/P53 signaling / 中国药理学与毒理学杂志

OBJECTIVE Chronic kidney disease (CKD) has become a global public health problem with 10％-15％incidence rate, and inhibiting the renal interstitial fibrosis is considered to be a potential strategy to delay the progression of CKD. Z-Guggulsterone (Z-GS), an active compound from derived from Commiphora mukul, has been proved to be effective in various diseases. The present study aimes to determine the protective effect and the molecular mechanism of Z-GS on renal fibrosis. METHODS Unilateral ureteral obstruction (UUO) mice and hypoxia-induced HK-2 cells were used to simulate renal fibrosis in vitro and in vivo, respectively. The mice and cells were treated with different doses of Z-GS to observe the pharmacological action. Renal function, including Scr, BUN, and UA, were detected by commercial kits. H&E and Masson staining were performed to observe histopathological changes of kidney. Cell viability and LDH release of HK-2 cells were detected by commercial kits. Cell cycle distribution and apoptosis rate were analyzed by flow cytometry. Fibrosis markers were detected by immunohistochemistry and immunofluorescence analysis. Cell cycle related proteins and Klotho/p53 signaling were analyzed by Western blotting. RESULTS The results showed that Z-GS decreased the rise of Scr, BUN, and UA and lightened renal histopathological injury, which were induced by UUO. Besides, Z-GS administration alleviated renal fibrosis in mice by inhibiting the expressions of α-SMA, TGF-β and colla?genⅣ, and delayed G2/M cell cycle arrest by promoting the expressions of CDK1 and cyclinD1/B1 rate. Experiments in vitro indicated that Z-GS treatment significantly increased the cell viability while decreased the LDH release in hypoxia-induced HK-2 cells. In addition, hypoxia induced fibrosis and G2/M cycle arrest in HK-2 cells were retarded by Z-GS. The study of its possible mechanism exhibited that Z-GS treatment increased the level of Klotho and inhibited P53 level. Nev?ertheless, the effect of Z-GS on Klotho/P53 signaling was reversed by siRNA-Klotho. Moreover, siRNA-Klotho treatment eliminated the effects of Z-GS on G2/M cell cycle arrest and fibrosis. CONCLUSION This study clarified that Z-GS allevi?ated renal fibrosis and G2/M cycle arrest through Klotho/P53 signaling pathway. People who have suffered CKD may potentially benefit from treatment with Z-GS.

Correlation between gut microbiota and FGF21 in mice with insulin resistance / 中国药理学通报

Aim To study the effect of normal glucose tolerance fecal microbiota transplantation ( FMT) on the gut microbiota in mice with insulin resistance(IR) and its correlation with FGF21 , and to explore the possible mechanisms of gut microbiota affecting IR.Methods After the establishment of IR model with high-fat diet feeding, 30 successful IR model mice were randomly divided into three groups; insulin resistance ( IR ) group, IR + metformin( Met) group, and IR + glucose tolerance normal fecal microbiota transplantation (FMT)group, and blank control(Control) group, with 10 mice in each group.After eight weeks of administration, the body mass and fasting blood glucose of mice at 8th week were recorded, then the number of target bacteria in fecal samples and the mRNA expression levels of FGF21 and its receptors in liver, colon and ileum tissues were detected by Real-time quantitative PCR( RT-qPCR).Results ® Compared with control group, the body mass and fasting blood glucose increased in IR group mice, while the mRNA expression levels of FGF21/p-Klolho/FGFRl/FGFR4 in liver, colon and ileum tissues were down-regulated.The levels of Bacteroules and R.sarlorii were reduced in fecal samples, and the levels of P.distasonis, M.schaedleri and R.gnavus increased.These indices were reverted by Met and FMT treatment.(2) The expression of FGF21 was negatively correlated with FBG, P.distasonis , M.schaedleri and R.gnavus, and positively correlated with Bacteroides and B.sartorii.Conclusions FMT can increase the expression level of FGF21 and regulate gut microbiota, and the two are closely related , which may be one of the important mechanisms of FMT in improving insulin resistance.

Effect of Klotho on oxidized low density lipoprotein-induced endothelial cell senescence via activation of autophagy / 解放军医学杂志

Objective: To investigate the effect of Klotho on oxidized low density lipoprotein (ox-LDL)-induced human coronary artery endothelial cell (HCAEC) senescence via activation of autophagy. Methods: HCAECs were cultured in vitro, and divided into control group and ox-LDL group [treated with ox-LDL at different concentrations (0, 25, 50, 75, 100 μg/ml) or different durations (0, 24 h, 48 h, 72 h, 96 h) 2 h after pre-incubation with Klotho (400 pmol/L)]. Cell viability was tested by CCK-8, senescent cells were detected by SA-β-GAL staining, immunofluorescence was used to detect the intracellular expression of P53 and P16, and Western blotting was performed to quantitatively analyze the P53, P16, LC3 and P62 protein expression. HCAECs were pre-incubated separately with Klotho (400 pmol/L), rapamycin (5 μmol/L) or chloroquine (2 μmol/L) for 2 hours under normal culture condition, and then cultured under ox-LDL exposure (75 μg/ml) for 72 hours. Cell viability was tested by CCK-8, senescent cells were detected by SA-β-GAL staining, autophagic flux was assayed by adenovirus GFP-LC3, and Western blotting was performed to quantitatively analyze the P53, P16, LC3, and P62 protein expression. Results: The results of CCK-8 assay, SAKlotho β-GAL staining and Western blotting respectively showed that Klotho decreased ox-LDL-induced inhibition of HCAECs viability (P<0.05), reduced the proportion of senescent cells (P<0.05), and down-regulated the expression of aging-related proteins P53 and P16 (P<0.05). Western blotting revealed that ox-LDL decreased LC3-II/LC3-I ratio and increased P62 protein expression level in a time-dependent (P<0.05) or dose-dependent manner (P<0.05) to some extent. Furthermore, Klotho increased LC3-II/LC3-I ratio (P<0.01), decreased P62 protein expression level (P<0.01), and reduced the inhibition of autophagy by chloroquine under ox-LDL exposure (P<0.05). The results of CCK-8 assay, SA-β-GAL staining and Western blotting respectively indicated that the HCAECs viability (P<0.01), the proportion of senescent cells (P<0.01), and the protein expression levels of P53 (P<0.01) and P16 (P<0.01) were significantly alleviated in ox-LDL+Klotho group and ox-LDL+rapamycin group than those in ox-LDL group; while the HCAECs viability (P<0.01), the proportion of senescent cells (P<0.01), and the protein expression levels of P53 and P16 (P<0.05) were obviously worse in chloroquine+ox-LDL group than those in ox-LDL group. Conclusion: Klotho ameliorates ox- LDL-induced HCAEC senescence via activation of autophagy.

Improvement effect of Shenyuan Granule on vascular calcification in db db diabetic nephropathy mice and its mechanism / 吉林大学学报(医学版)

Objective: To investigate the improvement effect of Shenyuan Granule on the vascular calcification in the db/db diabetic nephropathy (DN) mice induced by high phosphorus and its possible mechanism. Methods: Twenty SPF db/db mice were randomly divided into model group ( n= 10) and Shenyuan Granule group (n=10), and the wild type (WT) mice were chosen as blank group (?i=10). After treated for 12 weeks∗ the serum∗ kidney tissue and thoracic aorta tissue of the mice in various groups were taken. The serum FGF23 levels of the mice in various groups were detected by ELISA method; HE and von kossa staining were used to detect the pathomorphology of the thoracic aorta and calcium deposition of the mice in various groups; Real-time PCR method was used to detect the expression levels of Klotho mRNA in kidney tissue and Pit-1, Runx2 and SM22a mRNA in thoracic aorta tissue of the mice in various groups; Western blotting method was used to detect the expression levels of Klotho protein in kidney tissue and Pit-1, Runx2 and SM22a proteins in thoracic aorta tissue of the mice in various groups; immunohistochemisty method was used to detect the expression levels of Pit-1 protein in thoracic aorta tissue of the mice in various groups∗ and immunofluorescence method was used to detect the expression levels of SM22a and Runx2 proteins in thoracic aorta tissue of the mice in various groups. Results: Compared with blank group, the serum FGF23 level of the mice in model group was significantly increased ( P<0. 05), the expression levels of Klotho mRNA and protein in kidney tissue were significantly decreased ( P

Klotho: a potential protective protein for renal graft / 器官移植

Klotho gene is an anti-aging gene that is highly expressed in the kidney. Its encoding product Klotho protein can inhibit inflammation, oxidative stress injury, and apoptosis in renal tissue. It is regarded as a renal protective protein and expected to be a new target for the treatment of renal diseases. This article reviewed the biological characteristics of Klotho and the protective effect of Klotho on renal graft function.