(Hypertension. 2000;36:296.)
© 2000 American Heart Association, Inc.
Scientific Contributions |
From the Welch Center for Prevention, Epidemiology, and Clinical Research, Johns Hopkins University, Baltimore, Md (J.D.M., L.J.A., J.C); Department of Cardiology, Cairo University (Egypt) (Z.A., M.M.I.); and Tulane University School of Public Health and Tropical Medicine, New Orleans, La (P.K.W.).
Correspondence to Lawrence J. Appel, MD, MPH, Johns Hopkins University, 2024 E Monument St, Suite 2-645, Baltimore, MD 21205-2223. E-mail lappel{at}welch.jhu.edu
| Abstract |
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Key Words: skin color spectrophotometry blood pressure race
| Introduction |
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One explanation for the association of skin color and blood pressure (BP) is an imbalance in the distribution of factors that affect BP. Epidemiological studies have identified a number of variables associated with BP levels, including age, gender, body mass index (BMI), sodium intake, potassium intake, socioeconomic status, alcohol consumption, and physical activity.4 5 Whether these factors account for the increased rates of hypertension in blacks remains controversial.
A common gene that determines both skin color and BP is another explanation. The genetic mechanisms controlling melanin biosynthesis are complex and poorly understood. In the mouse, >50 distinct loci have been identified.6 Kwon7 has proposed that these genes be divided into 2 families: the tyrosinase family and the pmel 17 gene family. Human genes belonging to this first family include GP75 (location: 9p23), TYRP2 (location: 13q31-q32), and TYR (location: 11q14-q21). The primary gene belonging to the pmel 17 family is located at 12q13-q14. Genes regulating melanocyte-stimulating hormone and its cellular receptor are located at 2p25 and 16q24.3, respectively.
Still, a genetic link between melanin biosynthesis and BP regulation has yet to be identified. Association and linkage studies have identified several candidate genes for elevated BP: AGT (location: 1q42-q43), ACE (location: 17q23), REN (location: 1q32), and SAH (location: 16p13.11).8 Hence, at least for the present, the genes that might regulate BP do not appear to be linked to those regulating melanin biosynthesis.
A third mechanism explaining an association between skin color and BP is a common biochemical intermediate. Empirical data suggest that a common biochemical intermediate is plausible. For instance, Lerner et al9 found that administration of melanocyte-stimulating hormone to study participants resulted in increased pigmentation and, in some cases, increased diastolic blood pressure (DBP) and systolic blood pressure (SBP) levels. In addition, AGTI, a gene for a melanocyte-stimulating hormone antagonist located at 20q11.2, is related to skin pigmentation, as well as a predisposition to obesity, cardiovascular disease, and diabetes.10
In this setting, we hypothesized that BP rises with increasing levels of skin pigmentation and that this relationship is independent of other known risk factors for hypertension. To test this hypothesis, we used data from the Egyptian National Hypertension Project (NHP). In primary analyses, we used an objective measure of skin color, the Melanin Index, which is a continuous variable derived from reflectance spectrophotometry. In secondary analyses, we used a subjective measure of skin color in which participants were classified as black if they appeared to be Nubian in origin.
| Methods |
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140 mm Hg, a
mean DBP
90 mm Hg, or use of antihypertensive drug therapies
were defined as hypertensive. Phase II was a follow-up examination of
2313 individuals, ie, 1559 hypertensives identified in phase I and a
random sample of their normotensive counterparts (1 normotensive per 2
hypertensives, group-matched on gender and enumeration area). The phase
II visit occurred approximately 1 week after the phase I visit. The
study was approved by the institutional review board of the Egyptian
Ministry of Health. Consent was obtained from all
participants. In phase II, skin color measurements were obtained with the use of a portable spectrophotometer on all but 8 of the 1102 individuals living in Sharkia, Aswan, and El Wadi El Gedid. El Wadi El Gedid represented an exclusively urban-dwelling population, while approximately half of the participants in the other 2 governates were rural dwellers. From this group, 15 were excluded because of biologically implausible spectrophotometric data, 50 as a result of missing data related to the covariates of interest, 6 who were subjectively classified as having "white" skin because they were such a small number, and 188 because they were being treated with antihypertensive medications. After these exclusions, 835 persons (370 men and 465 women) were available for analysis.
Blood Pressure
BP was recorded according to a standard protocol adapted
from the American Heart Associations recommended
procedures12 by trained observers using a mercury
sphygmomanometer. Four measurements were obtained in phase I of the
study, and 2 additional readings were obtained in phase II of the
study. The measurements were taken at 5-minute intervals while
participants rested quietly in the seated position. In our
analyses, the primary outcome variables were SBP and DBP,
calculated as an unweighted average of the mean phase I and mean phase
II readings.
Skin Color Measures
For each participant, a spectrophotometric analysis of
skin color was performed with the use of a Minolta spectrophotometer
CM-2002. Measurements were taken from the mid-inner aspect of
the upper arm, an area unexposed to sunlight. For each individual, the
percentage of back-reflected light over the entire visible spectrum
(wavelengths between 400 and 700 nm) in 10-nm increments was
determined. Reflectance values ranged from 0 (none) to 1 (complete) at
each wavelength.
A Melanin Index, derived from the reflectance spectrum, was used as the primary measure of skin color. Derivation of this index was based on principles and methods as described by Dawson et al13 and Kollias and Baqer.14 In brief, it has been proposed that taking the log of the inverse reflectance at each wavelength transforms the remittance spectrum into a close approximation of the absorption spectrum. For light with wavelengths between 620 and 700 nm, log of the inverse reflectance values decrease in an approximately linear fashion with a slope proportional to the concentration of melanin in the skin. This slope is defined as the Melanin Index, a unitless, continuous variable objectively quantifying skin color. To facilitate its use, values were multiplied by (-1x105). In this study, large values of this index are indicative of dark skin.
A subjective measure of skin color was also recorded. Individuals were classified, on the basis of visual inspection by a study interviewer, as either "white," "fair," or "black." The classification took into account both the color of the skin and the racial origin of study participants. The black participants were composed of dark-skinned Egyptians, predominantly Nubians living in the Aswan governate.
Other Covariates
Basic demographic, socioeconomic, and health behavior
information was obtained from a questionnaire administered by a trained
interviewer. From a 12-hour overnight urine collection, sodium and
potassium excretions were determined, and the sodium-to-potassium ratio
was calculated. Subjects were classified as smokers if they gave a
positive response to the question "Do you currently smoke?" Weight
was measured with a double-beam scale placed on a firm surface, and
height was measured with a Frankfort plane positioned at a 90-degree
angle against a wall-mounted metal tape. BMI was calculated as weight
divided by height squared (kg/m2).
Three categories of education were none, primary (through primary school), and secondary (through or beyond secondary school). Occupation was classified as manual labor and nonmanual labor. Most manual laborers were farmers.
Statistical Methods
In multiple linear regression analyses, SBP and DBP were
the outcome variables. Multivariate model building
approaches included backward elimination and maximum
r2 methods using all variables
that showed a statistically significant association with BP, after
adjustment for age. Regression model diagnostics included
examining the residuals for normality, inspecting partial regression
plots, and examining the change in magnitude of the
r2 terms. All continuous variables
were standardized to have a mean of 0 and an SD of 1. Thus, the ß
coefficients represented the change in BP per SD increase
in these variables. All regression models included age, entered as
a series of "dummy" variables representing 10-year
age groups. Regression analyses were performed separately for
men and women because of a previously reported interaction between
gender and age on SBP.15
| Results |
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The mean age of the 465 women was 47.2 years. Almost half of the women (48.4%) came from Aswan. Women, compared with men, tended to have lower levels of education; 71.8% had no education, and only 11.2% had a secondary level of education. Educational levels were lowest in Aswan (79.6% with no education) and highest in El Wadi El Gedid (50.5% with no education). In general, women did not participate in the work force. Only 3.7% of women performed manual labor. The mean DBP and DBP for women were 134.6 and 80.1 mm Hg, respectively, and their mean BMI was 28.3 kg/m2. All but 1 of the 72 black women came from Aswan.
Figure 1 displays the distribution of Melanin Index for women. The distributions for both women and men are approximately gaussian with a right tail. The average Melanin Index value was 85.4 (range, 12.0 to 212.0) for women and 111.2 (range, 3.2 to 261.0) for men. Among those identified as being black-skinned and fair-skinned individuals, there was considerable overlap in Melanin Index values, primarily in the upper range.
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The users of antihypertensive medications excluded from the analyses differed from the study population in several respects. For both genders, those taking antihypertensive medications were more educated (P<0.05 for men and women) and had higher BMI values (P<0.001 for both) than the study participants. Fewer men taking antihypertensive medications were manual laborers (P<0.01), and fewer women taking medications were black (P<0.001). Among women, the majority of those taking antihypertensive medications were from El Wadi El Gedid, and the lowest proportion were from Aswan. Among men, the medication users overrepresented Sharkia and underrepresented Aswan, compared with the study participants. Finally, medication users had significantly lower Melanin Index values than the study participants. Male medication users had an average Melanin Index of 87.7 (P<0.0001), and female medication users had an average Melanin Index of 68.5 (P<0.0001).
Table 2 displays gender-specific bivariate relationships of Melanin Index to factors commonly associated with BP (age, BMI, sodium-to-potassium ratio, education, and occupation). Among men, the Melanin Index was inversely associated with BMI (r=-0.36, P<0.0001), education (P<0.001), and occupations other than manual labor (P<0.001). For women, the Melanin Index was positively correlated with age (P=0.02) and inversely correlated with BMI (P<0.0001) and sodium-to-potassium ratio (P=0.01). In men, manual laborers had darker skin than other occupational groups (P<0.001). Mean Melanin Index levels decreased across increasing educational strata for men and women (P<0.001).
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Higher levels of education were associated with younger age (P<0.001 for men and women), higher BMI (P<0.001 for men and women), lower Melanin Index values (P<0.001 for men and women), urban dwelling (P<0.001 for men and women), fair skin (P<0.001 for men; P<0.01 for women), and, for men, performing nonmanual labor (P<0.001).
Table 3 displays for men and women results from age-adjusted regression analyses in which BP was modeled as a function of 6 factors, each analyzed separately. In men, higher BMI (P=0.003) and secondary education (P=0.02) were significantly associated with higher levels of SBP. Manual laborers tended to have lower BP than other types of workers (P=0.07 for SBP and P=0.02 for DBP). Elevated DBP was significantly associated with higher BMI levels (P<0.0001) and secondary education (P=0.01). Higher levels of the Melanin Index were associated with lower DBP (P=0.03).
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In women, the relationship between Melanin Index and BP was nonlinear (Figure 2 and Table 3). Specifically, the largest increases in BP per unit increase in the Melanin Index occurred in the low to mid ranges of the Melanin Index; the relationship was attenuated in the upper ranges, where there were fewer observations. Because of the nonlinear association between the Melanin Index and BP, a squared term was added to the Melanin Index model for women. In contrast to men, completion of a secondary level of education was associated with lower levels of SBP (P=0.02) and DBP (P=0.006).
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Table 4 shows the results of multivariate linear regression analyses of SBP and DBP, separately for men and women. Factors considered for inclusion in this model were the covariates listed in Table 3. For men, age and the Melanin Index were forced into the models. Neither SBP nor DBP was significantly associated with Melanin Index in these models.
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For women, age was forced into both regression models. A significant nonlinear association between SBP and the Melanin Index persisted after adjustment for BMI and education. A similar pattern was seen with DBP, although the regression coefficient for the squared term was of borderline significance (P=0.09). When education was removed from the model, the regression coefficients for the Melanin Index and the Melanin Index squared terms in the DBP model were 2.1 (P=0.001) and -0.8 (P=0.05), respectively. When the multivariate analyses were stratified by governate, a significant (nonlinear) association between the Melanin Index and BP was observed only in Sharkia. We did not observe any interactions between educational level, skin color, and BP.
In separate analyses, we explored associations between BP and the subjective assessment of skin color (black [predominantly Nubian] versus fair). In these multivariate regression models, the analyses were restricted to residents of the Aswan governate because virtually all blacks in this study resided in this governate. The subjective skin color measure variable (classified as Nubian versus non-Nubian) was forced into all models. For both men and women, there was no significant difference between either SBP or DBP between Nubians and non-Nubians.
| Discussion |
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A major strength of this study was the use of an objective skin color measure that was proportional to the concentration of cutaneous melanins. Previous studies using spectrophotometric data measured skin color at specific wavelengths within the visible spectrum. In contrast, the Melanin Index used in this study measured skin color in the ranges where melanins are the primary light-absorbing compounds. Another strength was the availability of a large, well-characterized study population with a wide distribution of BP and skin color. Potential limitations include the cross-sectional design of the study and the lack of data on certain covariates (eg, measures of diet other than sodium and potassium and satisfactory measures of physical activity).
Five previous studies have examined the relationship between skin color and BP using a continuous measure of skin color derived from reflectance spectrophotometric data. The findings across these studies have been inconsistent. Boyle16 was able to demonstrate a graded, age-adjusted linear increase in both SBP and DBP with increasing skin darkness among black men and women participating in the Charleston Heart Study. However, in 2 follow-up studies of this cohort, Keil et al17 18 found no association between skin color and the incidence of hypertension after controlling for either education or other measures of social class. After adjusting for known hypertension risk factors, Klag et al19 found a positive linear relationship in both SBP and DBP only among black men and women of low socioeconomic status. Finally, Ernst et al20 found a significant association between skin darkness and SBP in black men and women. This relationship did not persist when the results were stratified by gender, causing the authors to postulate that their findings were most likely an artifact of BP and skin color differences between men and women.
Three hypotheses may explain a positive dose-response relationship between BP and skin pigmentation. For the first, skin color is interpreted as a marker for the degree of black-white admixture. Implicit in this hypothesis are the following assumptions: (1) genes exist that predispose an individual to higher BP; and (2) the prevalence of these genes is higher among black than white individuals. The first assumption is based on empirical observations of higher BP among blacks than among whites.1 Skin reflectance is a continuous measure of the degree of racial admixture between these populations,21 and individuals with the darkest skin have the highest proportion of genes from a black population, including the genes that confer increased risk for high BP. One would expect, therefore, to observe higher BP associated with increased levels of skin pigmentation.
The applicability of this model to the Egyptian population is limited on both theoretical and empirical grounds. First, if genes for melanin regulation and BP were linked, successive matings within an admixed population would diminish a linkage disequilibrium after several generations. Admixture in Egypt has likely been occurring for many generations among many different racial groups, thereby increasing the likelihood of a dilution. Second, skin color in Egypt represents admixture among several racial groups, including dark-skinned individuals not of West African origin. Thus, the assumption that the prevalence of genes predisposing to high BP is higher in those with darker color skin may not be tenable, particularly in light of the fact that BP in the Egyptian Nubian population, an ethnic group with physical characteristics similar to those of black Africans, did not significantly differ from that of the nonblack study participants. Consequently, it is not clear whether the assumptions of the admixture hypothesis are tenable.
A second explanation of a relationship between melanin levels and BP is a direct physiological association. The regulation of both BP and melanin synthesis is under hormonal control, and neither is independent of other systems.22 A mechanism of interaction, either through a specific gene product or a common metabolic intermediate, is not currently known.
Finally, a third explanation is confounding, namely, that skin color is associated with environmental factors known to influence BP. In our study, skin color was correlated with several variables that were independently associated with BP, including body mass, sodium-to-potassium excretion ratio (in women), and manual labor (in men), which one might consider a proxy for physical activity. However, in many but not all instances, the direction of the associations would tend to dilute a positive association between the Melanin Index and BP. For instance, darker-skinned women had lower BMIs and lower sodium-to-potassium excretion values than their lighter-skinned counterparts. Thus, one might expect dark skin to be associated with low BP.
Skin color was also associated with socioeconomic measures. For both genders, darker-skinned individuals had lower levels of education than their lighter-skinned counterparts. Among males, higher education was associated with higher BP, while the association was in the opposite direction for females. However, education may be an unsatisfactory proxy for socioeconomic status in this population, particularly among women; additionally, it is prone to residual confounding. Cohort effects were observed in both genders; the average age within educational strata decreased as the level of education increased, suggesting that advanced schooling is a recent phenomenon.
Skin color was also associated with antihypertensive medication use. Lighter-skinned men and women with high BP were more likely to use medications and therefore were more likely to be excluded from the study than darker-skinned individuals with high BP. The positive relationship between skin color and BP in women is consistent with the direction of the bias that may have been introduced through differential exclusions with respect both to skin color and BP. Furthermore, the association between skin color and medication use suggests that access to health services, another proxy for socioeconomic status, was more available to lighter-skinned individuals.
In summary, this study documented a significant relationship between skin color and BP in women but not in men. While the relationship appeared to be independent of other known risk factors for hypertension, the concomitant association of skin color with such variables as BMI, sodium-to-potassium excretion ratios, manual labor, and medication use suggests that residual confounding may explain the apparent association.
| Acknowledgments |
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Received November 10, 1999; first decision December 20, 1999; accepted February 28, 2000.
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