Introduction
Telomeres are DNA and protein complexes that are located at the end of linear chromosomes and are necessary for the complete replication of DNA as well as chromosome stability. Intact telomeres protect chromosomes from nuclease degradation, end-to-end fusion, and cellular senescence (Blackburn
2000). In general, telomeres shorten with age and can serve as an early predictor of the onset of several diseases, including hypertension (Demissie et al.
2006), atherosclerosis (Samani, Boultby, Butler, Thompson, and Goodall
2001), type 2 diabetes mellitus (Sampson, Winterbone, Hughes, Dozio, and Hughes
2006; Zee, Castonguay, Barton, Germer, and Martin
2010), cancer mortality (Willeit et al.
2010), cardiovascular disease (Epel et al.
2009a,
b), and cognitive decline and dementia (Martin‐Ruiz et al.
2006).
Interestingly, from a psychological perspective, telomere shortening can be accelerated by several behavioral factors, including poor diet (Nettleton, Diez-Roux, Jenny, Fitzpatrick, and Jacobs
2008; Valdes et al.
2005), poor sleep (Prather et al.
2011), cigarette smoking (Valdes et al.
2005), excessive alcohol consumption (Pavanello et al.
2011), sedentary lifestyle (Cherkas et al.
2008), and several psychological factors, such as personality characteristics (O’Donovan et al.
2009), psychiatric disorders (Lindqvist et al.
2015), and psychological distress (Shalev et al.
2013). For example, a reference study (Epel et al.
2004) compared the lengths of the telomeres in the white blood cells of mothers of chronically ill children with the telomere lengths of mothers of healthy children. The longer a woman had been the primary caregiver for her ill child (the children’s conditions ranged from gut disorders to autism), the shorter were her telomeres. Moreover, in both groups, the more severe a woman’s psychological stress was, the shorter her telomeres were. The reduction in telomere lengths of the most stressed mothers (compared with the reduction in the least stressed mothers) was equivalent to that caused by at least a decade of ageing.
It has been suggested that healthy lifestyle factors can promote telomerase (the cellular enzyme that adds telomeric repeat sequences to the ends of chromosomal DNA, preserving not only telomere length but also healthy cell function and long-term immune function), telomere maintenance or even lengthening. These factors include physical exercise (Puterman et al.
2010), a body mass index <25 kg/m
2 (Sun et al.
2012), not smoking (O’Donnell et al.
2008), and healthy diet (Paul
2011).
Meditation has also been proposed to be a healthy lifestyle factor that affects telomere length. Recent empirical studies have shown a positive association between meditation and longer telomeres (Hoge et al.
2013a,
b) as well as an increase in telomerase (Schutte and Malouff
2014), suggesting that meditation may play an important role in preventing illnesses. However, one of the most challenging questions to answer is how the practice of meditation is related to telomere dynamics.
The aim of this study was first to replicate and strengthen the hypothesis that meditation is associated with longer telomeres. Only a few studies have demonstrated this relationship (Schutte and Malouff
2014; Epel et al.
2009a,
b; Conklin et al.
2015; Jacobs et al.
2011). Second, to elucidate the psychological mechanisms underlying this potential relationship, we compared a group of expert meditators with a matched comparison group and used several questionnaires to assess different psychological constructs related to meditation that could be involved in delaying the ageing process. To measure telomere length, we employed the HT Q FISH technique, a unique method that not only determines the MTL but also the percentage of short telomeres in individual cells, which indicates the extent of cellular ageing.
Method
Measurements
Quantitative image acquisition and analysis was performed on a High Content Screening Opera System (Perkin Elmer) using the Acapella software, version 1.8 (Perkin Elmer, Waltham, MA, USA). Images were captured using a ×40 0.95 NA water immersion objective. UV and 488-nm excitation wavelengths were used to detect the DAPI and A488 signals, respectively. The results of the intensity of each foci identified were exported from the Acapella software (Perkin Elmer). The telomere length distribution and median telomere length were calculated with a proprietary program from Life Length.
Blood was obtained by venipuncture from the antecubital vein and collected into Sodium Heparin tubes. The blood samples were processed within 24 h of extraction to isolate peripheral blood mononuclear cells (PBMCs) by a Ficoll-Hypaque gradient. The PBMCs were rinsed in phosphate buffer solution, counted and resuspended at 10 million cells per milliliter in a freezing medium. Aliquots were frozen at −80 °C and placed in liquid nitrogen for storage.
Resilience was evaluated using the ten-item Connor-Davidson Resilience Scale (CD-RISC) (Campbell-Sills and Stein
2007), a self-administered questionnaire that is designed as a Likert type additive scale with five response options (0 = never to 4 = almost always), which has a single dimension in the original version. The final score on the questionnaire is the sum of the responses obtained from each item (range, 0–40), and the highest score indicates the highest level of resilience. The Spanish version of the scale has been recently validated and showed appropriate psychometric parameters (Notario-Pacheco et al.
2011) (Cronbach’s α = 0.85).
Life satisfaction was measured using the Satisfaction with Life Scale (SWLS) (Diener et al.
1985). The scale consists of five statements (e.g., “In most ways, my life is close to my ideal”), and the participants rate whether they agree or disagree with each statement using a five-point Likert scale. Overall life satisfaction (using a scale from 5 to 25) is calculated as the sum of the responses to all of the items; a higher score indicates a higher level of life satisfaction. Previous studies have consistently demonstrated a high reliability and a single-factor structure of SWLS items. This instrument has a translated and validated Spanish version that has shown adequate parameters (Vázquez et al.
2013) (Cronbach’s α = 0.86).
Subjective happiness was measured using the SHS (Lyubomirsky and Lepper
1999), which is a four-item measure of subjective global happiness rated on a seven-point Likert scale. The current study used the Spanish version of the SHS, which has established validity (Extremera and Fernández-Berrocal
2014). A single SHS score is the mean of the responses to the four items. The SHS scores can range from 1 to 7, where a higher score indicates a higher level of happiness (Cronbach’s α = 0.68–72).
Data Analyses
First, depending on their nature, all of the variables were described using either mean and standard deviation (SD) values or percentages. The comparisons between groups were performed using Student’s
t test and a chi-squared test. The effect sizes on telomere length between meditators and nonmeditators were assessed using Cohen’s
d. The descriptive statistics and raw Pearson correlations (
r) between the sociodemographics, the psychological variables, and the two telomere measurements were calculated considering the total sample. The sociodemographic and psychological variables that showed significant results in the raw analysis were included in two multivariate linear regression models that assessed the adjusted relationships between the telomere measurements and psychological outcomes. We used the stepwise method to introduce the variables into the regression models and to assess the adjusted relationships between the telomere measurements and psychological outcomes because of the small sample size (
n = 40) and the subsequent problems with statistical power. Standardized coefficients (beta) were used to assess the individual contribution of the variables in explaining the telomere length, and the Wald test was used to evaluate the significance of the variables. The adjusted multiple determination coefficients (adj-
R
2) were also calculated to observe their grouped explanatory power, and their significance was assessed using analysis of variance (Martínez-González et al.
2006). The partial correlation coefficients (
R) were calculated, which indicated the correlation between two variables when the effects of the other variables in the equation were removed. The Kolmogorov-Smirnov (KS) test was used to determine whether the conditional distribution of the residuals met the assumption of normality. Finally, it was confirmed that the Durbin-Watson values (DW) approached a value of approximately 2.00 to rule out autocorrelation problems in the errors. All of the tests used were bilateral, and the significance level was α < 0.05. SPSS-19 statistical software package was used.
Results
One meditator was excluded from the analyses because he had prostate cancer, and two individuals in the comparison group were excluded because they were being treated with antidepressants. The sociodemographic and health characteristics of the sample are summarized in Table
1. There was a predominance of middle-aged elderly European men. Gender, age, and ethnic group variables were matched to avoid significant differences. There were no differences between the samples in other sociodemographic variables (living with a partner and years of education), most health habits (tobacco, alcohol and medication consumption and vegetarian diet), and several chronic medical disorders. The only differences between the groups were in the average body mass index (BMI), which was significantly higher in the meditators and showed mild overweight (BMI >25), and the amount of physical exercise, which was lower in the meditators. The mean period of daily meditation reported by the meditators was 75 min with a standard deviation of 15 min. The mean length of time that the participants had practiced meditation over their lifetime was 180 months with a standard deviation of 12 months. Both of the measures were rounded by the participants because it was difficult to provide more accurate data, resulting in quite homogenous data.
Table 1
Sociodemographic and health characteristics of the sample
Sociodemographics |
Gender (male)a
| 14/20 (70 %) | 14/20 (70 %) |
X
2 = 0.0; df = 1; p = 1 |
Age (mean, SD)a
| 48.55 (8.05) | 48.30 (8.76) |
t = 0.094; df = 38; p = 0.926 |
Ethnic group (white)a
| 20/20 (100 %) | 20/20 (100 %) |
X
2 = 0; df = 1; p = 1 |
Live with partner (%) | 17/20 (85 %) | 18/20 (90 %) |
X
2 = 0.229; df = 1; p = 0.663 |
Years of education (mean, SD) | 14.25 (4.37) | 12.60 (3.11) |
t = 1.37; df = 38; p = 0.178 |
Healthy habits |
Tobacco (>10 cigarettes/day) | 3/20 (15 %) | 5/20 (25 %) |
X
2 = 0.625; df = 1; p = 0.429 |
Alcoholb
| 0/20 (0 %) | 0/20 (0 %) |
X
2 = 0; df = 1; p = 1 |
Medications | 0/20 (0 %) | 0/20 (0 %) |
X
2 = 0; df = 1; p = 1 |
Vegetarian diet | 0/20 (0 %) | 0/20 (0 %) |
X
2 = 0; df = 1; p = 1 |
Body mass index (mean, SD) | 25.25 (1.50) | 23.79 (1.96) |
t = 2.63; df = 38; p = 0.012* |
Exercise (>3 h/week) | 1/20 (5 %) | 7/20 (35 %) |
X
2 = 5.65; df = 1: p = 0.048* |
Medical disorders |
Diabetes | 1/20 (5 %) | 3/20 (15 %) |
X
2 = 1.111; df = 1; p = 0.292 |
Hypertriglyceridemia | 1/20 (5 %) | 3/20 (15 %) |
X
2 = 1.111; df = 1; p = 0.292 |
Hypercholesterolemia | 1/20 (5 %) | 3/20 (15 %) |
X
2 = 1.111; df = 1; p = 0.292 |
Hypertension | 3/20 (15 %) | 4/20 (20 %) |
X
2 = 0.173; df = 1; p = 0.677 |
Arthrosis | 1/20 (5 %) | 3/20 (15 %) |
X
2 = 1.111; df = 1; p = 0.292 |
The other health variables included in the Labco health questionnaire were not summarized in the table due to space constraints; however, they did not show significant differences between the groups. These variables included the prevalence of cardiovascular disorders (heart infarction, atherosclerosis, arrhythmia, and varicose veins), neurodegenerative disorders (Alzheimer’s disease, Parkinson’s disease, multiple sclerosis, and lateral amyotrophic sclerosis), infectious disorders (syphilis, hepatitis B and C, mononucleosis, toxoplasmosis, and cytomegalovirus), autoimmune disorders (psoriasis, rheumatoid arthritis, and lupus erythaematosus), and other disorders (osteoporosis, chronic obstructive pulmonary disease, lung fibrosis, congenital dyskeratosis, progeria, aplastic anaemia, asthma, and allergies). Other variables assessed were the use of different medications (such as oestrogens, thyroid hormone agents, and antioxidants) and characteristics of the participant’s diet (such as consumption of fruits, vegetables, red meat, and fat).
Telomere Measurement
A t test showed that the expert meditators group (mean = 10.82 kb; SEM = 0.23; SD = 1.03) had a significantly longer MTL (t = 2.97; df = 38; p = 0.005; Cohen’s d = 0.94) compared with the comparison group (mean = 9.94 kb; SEM = 0.19; SD = 0.84).
The data concerning the 20th percentile were similar, showing that the prevalence of short telomeres in the cells of the expert meditators group (mean = 5.22 kb; SEM = 0.11; SD = 0.48) was significantly lower (t = 2.84; df = 38; p = 0.007; Cohen’s d = 0.91) than the comparison group (mean = 4.80 kb; SEM = 0.10; SD = 0.44).
Mindfulness Variables
As expected, the expert meditators showed significantly better results in the measurements that were related to mindfulness abilities, such as attention and awareness, observing, describing, nonjudging, resilience, self-compassion, and satisfaction with life and subjective happiness. Moreover, the expert meditators reported significantly lower experiential avoidance, anxiety, and depression (Table
2).
Table 2
Psychological variables in meditators (n = 20) and nonmeditators (n = 20)
MAAS | 4.50 (0.16) | 0.09 | 0.15 | 3.39 (0.26) | 0.44 | 0.43 | 15.79 (38)*** | 0.49** | 0.49** |
FFMQ Observing | 28.65 (1.42) | 0.29 | 0.11 | 23.95 (0.99) | −0.10 | −0.05 | 12.08 (38)*** | 0.45** | 0.39* |
FFMQ Describing | 26.90 (1.33) | −0.46* | −0.42 | 29.25 (1.29) | −0.13 | −0.05 | −5.66 (38)*** | −0.50** | −0.45** |
FFMQ Acting aware | 32.55 (1.50) | −0.35 | −0.28 | 27.75 (1.44) | −0.01 | 0.12 | 10.28 (38)*** | 0.46** | 0.40* |
FFMQ Nonjudging | 31.65 (1.46) | 0.20 | 0.23 | 26.85 (4.46) | −0.15 | −0.06 | 4.57 (38)*** | 0.23 | 0.26 |
FFMQ Nonreacting | 22.95 (2.50) | −0.35 | −0.29 | 21.35 (2.41) | −0.07 | −0.06 | 2.05 (38)* | −0.05 | −0.02 |
AAQ2 | 14.75 (2.82) | −0.41 | −0.41 | 22.60 (1.69) | −0.23 | −0.09 | −10.64 (38)*** | −0.53*** | −0.49** |
HADS Anx | 0.75 (0.63) | 0.30 | 0.30 | 4.05 (0.75) | −0.48* | −0.47* | −14.87 (38)*** | −0.43** | −0.42** |
HADS Dep | 0.40 (0.59) | 0.12 | 0.27 | 4.05 (0.99) | 0.15 | 0.20 | −14.02 (38)*** | −0.35* | −0.30 |
CD-RISC | 31.10 (1.11) | −0.01 | −0.08 | 24.45 (1.50) | −0.28 | −0.26 | 15.86 (38)*** | 0.36* | 0.33* |
SWLS | 29.30 (2.55) | 0.07 | −0.03 | 23.00 (1.89) | −0.06 | −0.13 | 8.85 (38)*** | 0.37* | 0.31 |
SHS | 27.20 (2.21) | 0.31 | 0.33 | 21.10 (1.74) | 0.14 | 0.18 | 9.67 (38)*** | 0.48** | 0.48** |
SCS Self-kindness | 5.40 (0.68) | 0.27 | 0.19 | 4.45 (0.82) | 0.05 | 0.19 | 3.97 (38)*** | 0.36* | 0.37* |
SCS Humanity | 5.35 (1.08) | 0.40 | 0.37 | 3.90 (0.71) | 0.09 | 0.13 | 4.97 (38)*** | 0.47** | 0.46** |
SCS Mindfulness | 4.80 (1.10) | 0.08 | 0.12 | 3.75 (0.63) | −0.18 | −0.19 | 3.67 (38)*** | 0.22 | 0.23 |
Telomeres and Mindfulness Variables
Regarding the sociodemographic variables, only age showed significant relationships with MTL (
r = 0.66;
p < 0.001) and 20th percentile (
r = 0.64;
p < 0.001). The raw correlations between the two references of telomere measurements and the psychological variables supported the study’s hypothesis, as these variables were associated in the expected direction (Table
2). Of particular importance was the high correlation shown between telomere length and experiential avoidance (
r = −0.53;
p < 0.001). Notably, only three mindfulness subscales did not show a significant association with telomeres: nonjudging, nonreactivity, and self-compassion mindfulness. To determine which of the mindfulness variables was responsible for the telomere maintenance, we computed a regression analysis with regard to MTL (Table
3). We used a stepwise regression model (because of the limited statistical power) with the following variables: group (Zen/comparator), age (the only sociodemographic variable with a significant association with telomere length) and all of the psychological variables that showed a significant association with telomere length. As a result, only the variables that best explained the variability in telomere length were included in the final model, as the variables that did not add new information to the variables introduced in previous steps were removed.
Table 3
Regression models regarding the MTL and the 20th percentile (N = 40)
MTL | 0.73 | 35.78 (1/36) <0.001 | 0.54 | 2.03 | 0.923 |
R
| B (95 % CI) | Se | Beta |
p
c
|
Intercept | | 14.73 (12.77 to 16.69) | 0.97 | | <0.001 |
Age | −0.80 | −0.08 (−0.10 to −0.06) | 0.01 | −0.67 | <0.001 |
AAQ-II | −0.46 | −0.08 (−0.13 to −0.03) | 0.03 | −0.35 | 0.004 |
SCS Humanity | 0.37 | 0.23 ( 0.03 to 0.44) | 0.10 | 0.27 | 0.024 |
Variables | Adj-R2
|
F (df
1/df
2) p
a
| Se | DW |
p
b
|
20th percentile | 0.67 | 26.85 (1/36) <0.001 | 0.29 | 2.02 | 0.596 |
R
| B (95 % CI) | Se | Beta |
p
c
|
Intercept | | 6.94 ( 5.88 to 8.00) | 0.52 | | <0.001 |
Age | −0.76 | −0.04 (−0.05 to −0.03) | 0.01 | −0.65 | <0.001 |
AAQ-II | −0.37 | −0.03 (−0.06 to −0.01) | 0.01 | −0.29 | 0.024 |
SCS Humanity | 0.36 | 0.12 (0.01 to 0.23) | 0.05 | 0.29 | 0.029 |
Surprisingly, the components that measured mindfulness skills in the MAAS and FFMQ scales did not contribute to the final model. However, the following three factors made significant contributions: age (beta = −0.67;
p < 0.001), experiential avoidance (beta = −0.35;
p = 0.004), and the common humanity subscale from the self-compassion scale (beta = 0.27;
p = 0.024). Next, a second regression model was conducted using the 20th percentile telomere values as the reference. The results were similar to those shown in the first model. Age (beta = −0.65;
p < 0.001), experiential avoidance (beta = −0.29;
p = 0.024) and the common humanity subscale of the self-compassion scale (beta = 0.29;
p = 0.029) contributed significantly (Table
3).
Discussion
The present data demonstrated that the expert meditators had a significantly longer MTL as well as lower percentages of short telomeres in their cells than the nonmeditator comparison group. This finding extends a small but growing body of literature showing longer telomeres (Schutte and Malouff
2014; Epel et al.
2009a,
b; Conklin et al.
2015; Jacobs et al.
2011) and increases in telomerase (Lengacher et al.
2014) related to the practice of mindfulness. The ability to maintain longer telomeres through practicing meditation has many implications on health. The possible pathway between meditation and telomere length seems to be that (Schutte and Malouff
2014) mindfulness leads to individuals experiencing less stress, anxiety, and depression, which are all thought to be associated with cortisol level, and this association seems to be associated with telomerase activity.
Previous research has utilized the qPCR method for telomere measurement, which only provides the mean (not median) telomere length value per cell or per sample. However, a unique aspect of our study is that we applied the HT Q FISH method and were able to measure not only the MTL but also the abundance of short telomeres, which is an important parameter for assessing telomere functionality. Moreover, previous studies have shown similar results for the practice of Loving Kindness Meditation (Salzberg
1995), a type of meditation practice that focuses on developing a positive intention, unselfish kindness and warmth toward all people (Shaku et al.
2014). However, this study assessed a Zen Buddhism practice. Thus, it seems that mindfulness is a protective factor for telomere length regardless of the type of meditation practiced. These results might also be expected because Zen meditation has already been related not only to improvements in quality of life, better mental health (Shaku et al.
2014), and alpha and theta activity in many brain regions (generally related to relaxation) (Chiesa
2009) but also to decreases in oxidative stress (Mahagita
2010) and the resiliency of mitochondria (Bhasin et al.
2013), which may help prevent the process of ageing.
However, the exact drivers of mindfulness and their relationship to telomere length remain unclear. According to our results, it is likely that acceptance (measured as the absence of experiential avoidance by the AAQ-2), a process that is specifically promoted by mindfulness in general and Zen meditation in particular, plays a key role. However, there is still debate over which concept the AAQ-2 truly measures; some authors believe (Wolgast
2014) that it assesses distress and not avoidance/acceptance.
Acceptance has been found to predict a wide range of quality of life outcomes, such as depression, anxiety, and general mental health. It has been successfully applied to several specific topics, such as pain, smoking, diabetes management, tinnitus, weight, and coping with epilepsy and psychotic symptoms (Bond et al.
2011). However, this study is one of the first to show that acceptance is related to telomere length. Undoubtedly, from a cellular perspective, our results strengthen the idea that mental health and behavioural effectiveness are influenced more by how people relate to their thoughts and feelings than by the form of those thoughts or feelings (e.g., how negative they are).
Another factor that seems to play an essential role at the cellular level is one of the components of the self-compassion scale: common humanity. Persons with high anxiety have low mindfulness and compassion scores, and in these populations, mindfulness is a better predictor of disability than anxiety symptoms. This suggests that mindfulness can help protect against the feeling of being disabled by an anxiety disorder (Hoge et al.
2013a,
b). Common humanity can also be viewed as a useful emotional regulation strategy in which painful or distressing feelings are not avoided but are held in awareness with kindness, understanding, and a sense of shared humanity.
Finally, it is necessary to emphasize that nearly all of the psychological variables (MAAS, AAQ, FFMQ, HADS, CD-RISC, SCS) were correlated with the telomere measures. No particular psychological variable had an association with telomere length that was clearly stronger than that of the others. Socioeconomic status did not show any relationship with telomere length despite being considered a relevant variable. In our sample, the meditators had a significantly lower salary than the comparison group because some of them were Buddhist monks who had taken vows of poverty; this fact may have biased the data.
This study has a number of limitations that suggest a cautious interpretation of the results. The most significant limitation of this research was its small sample size. Second, the processes examined are technically complex to measure, and in many ways, the instruments used are recent developments. Additional experience may lead us to refine these instruments, and different or improved instruments may reveal a different pattern of results. In addition, the participants were not randomly assigned to groups; thus, causality is unclear. Finally, telomere research is still in its early stages and potential variables that are difficult to measure and are currently unknown may alter the results.
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