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Background
With the global aging of populations, there is a growing need for objective indicators that capture interindividual variability in the aging process. In recent years, biological age has been increasingly recognized as a more informative measure than chronological age, as it integrates multiple physiological and biochemical markers reflecting systemic function. The derived concept of biological age acceleration (BAA), defined as the difference between biological age and chronological age, has been proposed as an indicator of the rate of aging at the individual level. Within this context, considerable attention has been directed toward identifying circulating molecules that reflect biological aging processes. Among these candidates, α-Klotho, a protein widely recognized for its anti-aging properties, has emerged as a potential biomarker of aging (PLoS One. 2026; 21(3):e0343429.).

Overview of the Study
The present study utilized data from the National Health and Nutrition Examination Survey (NHANES 2007–2010), including 5,654 middle-aged and older adults aged 45 to 85 years. Serum α-Klotho concentrations were measured by enzyme-linked immunosorbent assay (ELISA), and biological age was estimated by validated composite biomarker algorithm (BioAge). BAA was calculated as the difference between biological age and chronological age. Multivariable regression models were applied with adjustment for a comprehensive set of covariates, including demographic variables, lifestyle factors, socioeconomic status, and comorbid conditions.

Results
The analysis demonstrated a significant inverse association between circulating Klotho levels and biological age acceleration across all statistical models. Higher levels of log-transformed Klotho were consistently associated with lower BAA, indicating a slower pace of biological aging. This inverse association remained robust after full adjustment for confounding factors. Consistent results were also observed in logistic regression analyses, where higher Klotho concentrations were associated with lower odds of accelerated biological aging, supporting the stability of the observed relationship across analytical approaches.

Importantly, the association between Klotho levels and BAA was found to be non-linear. Restricted cubic spline analysis revealed an L-shaped relationship, with an inflection point at approximately 791.85 pg/mL. Below this threshold, BAA declined sharply with increasing Klotho levels, whereas above this level the decline became more gradual.

Furthermore, the study demonstrated that the association between α-Klotho and BAA varied by individual background. The inverse association between Klotho levels and biological age acceleration was more evident among moderate alcohol consumers. In contrast, the presence of diabetes attenuated this association, with a weaker inverse relationship observed among individuals with diabetes compared with those without diabetes. These findings suggest that the association between Klotho and biological aging may be influenced by metabolic status and lifestyle-related factors.

Discussion and Future Perspectives
From a biological standpoint, Klotho is known to regulate multiple pathways implicated in aging. It functions as a co-receptor for fibroblast growth factor 23 (FGF23), thereby contributing to the regulation of phosphate and vitamin D metabolism. In addition, Klotho has been implicated in the modulation of oxidative stress and inflammatory responses, including activation of antioxidant pathways and suppression of pro-inflammatory signaling. Furthermore, Klotho interacts with insulin and IGF-1 signaling pathways, which are closely associated with longevity and metabolic regulation. These mechanisms are consistent with the observed associations between circulating Klotho levels and biological age acceleration.

These findings have potential implications for both research and clinical development. α-Klotho may serve as a biomarker of biological aging, reflecting the functional status of multiple physiological systems. In addition, previous studies have shown that circulating Klotho levels may increase in response to lifestyle interventions, such as physical activity and weight reduction, suggesting a possible role for Klotho in monitoring intervention effects. Furthermore, ongoing research into therapeutic strategies aimed at increasing Klotho levels, including recombinant protein administration and the enhancement of endogenous expression, may provide new approaches for the management of age-related conditions.

In conclusion, this large population-based analysis demonstrates a consistent inverse association between circulating α-Klotho levels and biological age acceleration. These results support the potential role of Klotho as a biomarker reflecting the rate of aging, while highlighting the need for longitudinal and interventional studies to clarify its causal role and clinical applicability.


Please refer to the following publication for more details.

Mengna Huang et al.
Klotho levels and biological age acceleration: Insights from a diverse cohort of middle-aged and elderly individuals
PLoS One. 2026; 21(3):e0343429.

We offer a wide range of ELISA kits for Aging research and also provide testing services.

■NHANES
The National Health and Nutrition Examination Survey (NHANES) is a semi-annual survey conducted by the Centers for Disease Control and Prevention (CDC) to monitor the health and nutritional status of the U.S. population.
On this survey, the CDC selected our ELISA kit below to measure serum α-Klotho levels in a total of more than 13,000 samples collected between 2007 and 2016. Please also refer to the related IBL news.

We hope this information will be useful for your research.
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