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Klotho preservation via histone deacetylase inhibition attenuates chronic kidney disease-associated bone injury in mice

View Article: PubMed Central - PubMed

ABSTRACT

Bone loss and increased fracture are the devastating outcomes of chronic kidney disease-mineral and bone disorder (CKD-MBD) resulting from Klotho deficit-related mineral disturbance and hyperparathyroidism. Because Klotho down-regulation after renal injury is presumably affected by aberrant histone deacetylase (HDAC) activities, here we assess whether HDAC inhibition prevents Klotho loss and attenuates the CKD-associated bone complication in a mouse model of CKD-MBD. Mice fed adenine-containing diet developed the expected renal damage, a substantial Klotho loss and the deregulated key factors causally affecting bone remodeling, which were accompanied by a marked reduction of bone mineral density. Intriguingly, administration of a potent HDAC inhibitor trichostatin A (TSA) impressively alleviated the Klotho deficit and the observed alterations of serum, kidney and bone. TSA prevented Klotho loss by increasing the promoter-associated histone acetylation, therefore increasing Klotho transcription. More importantly the mice lacking Klotho by siRNA interference largely abolished the TSA protections against the serum and renal abnormalities, and the deranged bone micro-architectures. Thus, our study identified Klotho loss as a key event linking HDAC deregulation to the renal and bone injuries in CKD-MBD mice and demonstrated the therapeutic potentials of endogenous Klotho restoration by HDAC inhibition in treating CKD and the associated extrarenal complications.

No MeSH data available.


Related in: MedlinePlus

HDAC inhibition restores declined renal Klotho in adenine mice.(A) Immunohistochemical staining. The kidney sections from control, TSA, adenine and TSA-treated adenine mice (6 weeks, n = 6) were examined by immunohistochemical staining for renal Klotho expression. The representative figures from each group were shown. (B) Serum Klotho levels. The average concentrations of serum Klotho from control, TSA, adenine and TSA-treated adenine mice were measured by ELISA (n = 6). (C) Bone Klotho mRNA. The relative levels of Klotho mRNA were examined from control, TSA, adenine and TSA-treated adenine mouse femurs by qRT-PCR (n = 6). (D) Renal expressions of Klotho, α-SMA and E-cadherin were assayed by Western blotting from the mouse kidneys (2 randomly selected samples were shown). (E) Quantifications of Fig. 3D (n = 6). Data are presented as mean ± SD. *P < 0.05 versus control, #P < 0.05 versus adenine mice.
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f3: HDAC inhibition restores declined renal Klotho in adenine mice.(A) Immunohistochemical staining. The kidney sections from control, TSA, adenine and TSA-treated adenine mice (6 weeks, n = 6) were examined by immunohistochemical staining for renal Klotho expression. The representative figures from each group were shown. (B) Serum Klotho levels. The average concentrations of serum Klotho from control, TSA, adenine and TSA-treated adenine mice were measured by ELISA (n = 6). (C) Bone Klotho mRNA. The relative levels of Klotho mRNA were examined from control, TSA, adenine and TSA-treated adenine mouse femurs by qRT-PCR (n = 6). (D) Renal expressions of Klotho, α-SMA and E-cadherin were assayed by Western blotting from the mouse kidneys (2 randomly selected samples were shown). (E) Quantifications of Fig. 3D (n = 6). Data are presented as mean ± SD. *P < 0.05 versus control, #P < 0.05 versus adenine mice.

Mentions: Klotho is a key gene profoundly affecting mineral metabolism and bone remodeling, but reportedly suppressed after renal injury. We then decided to test whether HDAC inhibition affects Klotho expression. The results showed that Klotho levels in kidney, blood circulation and femur were all markedly reduced in adenine mice, as demonstrated by immunohistochemical and Western blotting examinations of kidney tissues (Fig. 3A,D and E), ELISA assay of mouse sera (Fig. 3B) and quantitative real time PCR measurement of mRNA in femur. (Fig. 3C). However TSA treatment impressively attenuated the Klotho reductions in all assays. In addition, the typical CKD-associated pathological alterations, such as the reduction of E-cadherin- an epithelial cell marker and the induction of α-SMA, the myofibroblast differentiation marker, were also attenuated by TSA treatment, indicating that HDAC inhibition effectively prevents Klotho loss, which might account for the renal and bone protections.


Klotho preservation via histone deacetylase inhibition attenuates chronic kidney disease-associated bone injury in mice
HDAC inhibition restores declined renal Klotho in adenine mice.(A) Immunohistochemical staining. The kidney sections from control, TSA, adenine and TSA-treated adenine mice (6 weeks, n = 6) were examined by immunohistochemical staining for renal Klotho expression. The representative figures from each group were shown. (B) Serum Klotho levels. The average concentrations of serum Klotho from control, TSA, adenine and TSA-treated adenine mice were measured by ELISA (n = 6). (C) Bone Klotho mRNA. The relative levels of Klotho mRNA were examined from control, TSA, adenine and TSA-treated adenine mouse femurs by qRT-PCR (n = 6). (D) Renal expressions of Klotho, α-SMA and E-cadherin were assayed by Western blotting from the mouse kidneys (2 randomly selected samples were shown). (E) Quantifications of Fig. 3D (n = 6). Data are presented as mean ± SD. *P < 0.05 versus control, #P < 0.05 versus adenine mice.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC5384196&req=5

f3: HDAC inhibition restores declined renal Klotho in adenine mice.(A) Immunohistochemical staining. The kidney sections from control, TSA, adenine and TSA-treated adenine mice (6 weeks, n = 6) were examined by immunohistochemical staining for renal Klotho expression. The representative figures from each group were shown. (B) Serum Klotho levels. The average concentrations of serum Klotho from control, TSA, adenine and TSA-treated adenine mice were measured by ELISA (n = 6). (C) Bone Klotho mRNA. The relative levels of Klotho mRNA were examined from control, TSA, adenine and TSA-treated adenine mouse femurs by qRT-PCR (n = 6). (D) Renal expressions of Klotho, α-SMA and E-cadherin were assayed by Western blotting from the mouse kidneys (2 randomly selected samples were shown). (E) Quantifications of Fig. 3D (n = 6). Data are presented as mean ± SD. *P < 0.05 versus control, #P < 0.05 versus adenine mice.
Mentions: Klotho is a key gene profoundly affecting mineral metabolism and bone remodeling, but reportedly suppressed after renal injury. We then decided to test whether HDAC inhibition affects Klotho expression. The results showed that Klotho levels in kidney, blood circulation and femur were all markedly reduced in adenine mice, as demonstrated by immunohistochemical and Western blotting examinations of kidney tissues (Fig. 3A,D and E), ELISA assay of mouse sera (Fig. 3B) and quantitative real time PCR measurement of mRNA in femur. (Fig. 3C). However TSA treatment impressively attenuated the Klotho reductions in all assays. In addition, the typical CKD-associated pathological alterations, such as the reduction of E-cadherin- an epithelial cell marker and the induction of α-SMA, the myofibroblast differentiation marker, were also attenuated by TSA treatment, indicating that HDAC inhibition effectively prevents Klotho loss, which might account for the renal and bone protections.

View Article: PubMed Central - PubMed

ABSTRACT

Bone loss and increased fracture are the devastating outcomes of chronic kidney disease-mineral and bone disorder (CKD-MBD) resulting from Klotho deficit-related mineral disturbance and hyperparathyroidism. Because Klotho down-regulation after renal injury is presumably affected by aberrant histone deacetylase (HDAC) activities, here we assess whether HDAC inhibition prevents Klotho loss and attenuates the CKD-associated bone complication in a mouse model of CKD-MBD. Mice fed adenine-containing diet developed the expected renal damage, a substantial Klotho loss and the deregulated key factors causally affecting bone remodeling, which were accompanied by a marked reduction of bone mineral density. Intriguingly, administration of a potent HDAC inhibitor trichostatin A (TSA) impressively alleviated the Klotho deficit and the observed alterations of serum, kidney and bone. TSA prevented Klotho loss by increasing the promoter-associated histone acetylation, therefore increasing Klotho transcription. More importantly the mice lacking Klotho by siRNA interference largely abolished the TSA protections against the serum and renal abnormalities, and the deranged bone micro-architectures. Thus, our study identified Klotho loss as a key event linking HDAC deregulation to the renal and bone injuries in CKD-MBD mice and demonstrated the therapeutic potentials of endogenous Klotho restoration by HDAC inhibition in treating CKD and the associated extrarenal complications.

No MeSH data available.


Related in: MedlinePlus