With respect to the optimal macronutrient composition in the daily diet, most international authorities recommend to increase intakes of carbohydrates at the expense of fat and protein [1, 2]. However, in face of the worldwide increase in prevalence of both overweight and obesity, there is a plethora of recommendations for diets aiming at weight loss and weight management. Among them, a high-protein (HP) regimen has gained increasing interest in recent years [3]. For the general population, recommended dietary reference intakes (DRIs) for protein are 0.66 g * kg body weight^-1 * d^-1[4]. Actual consumption data for the US American population average 1.3 g * kg body weight^-1 * d^-1 in the 19–30 age group indicating a protein intake in excess of their needs [5]. The Acceptable Macronutrient Distribution Range (AMDR) for protein is given as 5-35% of daily calories depending on age [6]. A recent meta-analysis comparing HP vs. low-protein (LP) diets with a duration between 28 days and 12 months observed favorable effects of HP diets on biomarkers of obesity as well as cardiovascular risk factors such as HDL-cholesterol (HDL-C), triacylglycerols (TG), and blood pressure [7]. Several randomized controlled trials (RCTs) investigated the short-term effects of HP vs. LP diets, reporting advantages of HP protocols including a reduction in TG concentration [8–10]. A meta-regression of 87 studies concluded that low-carbohydrate, HP diets favorably affected body mass and composition independent of energy intake [11]. The benefits of HP diets might be explained by increased thermogenesis and satiety [12, 13]. Recent data from the 26-year follow up of the Nurses’ Health Study (NHS) revealed that protein sources such as red meat and high-fat dairy products were significantly associated with an elevated risk of coronary heart disease, while higher intakes of poultry, fish, and nuts correlated with a lower risk of coronary heart disease (CHD) [14]. Since there is a lack of information concerning studies with different protein contents covering a longer dietary intervention period, the aim of this meta-analysis was to compare the long-term effects of HP vs. LP regimens on biomarkers of obesity, cardiovascular complications as well as adverse effects of HP.

The review protocol has been registered in PROSPERO International Prospective Register of Systematic Reviews (crd.york.ac.uk/prospero/index.asp Identifier: CRD42012002791).

Quality assessment of studies

Full copies of studies were independently assessed for methodological quality by both authors using the Jadad score [17]. This 5-point quality scale includes points for randomization (randomized = 1 point; table of random numbers or computer generated randomization = an additional 1 point), double-blinding (double-blind = 1 point; use of a placebo = additional 1 point), and follow-up (numbers and reasons for withdrawal in each group are stated = 1 point) within the report of an RCT. An additional point was accepted if the analysis was by intention-to-treat to compensate for the fact that double-blinded study protocols are elusive in dietary intervention studies. Final scores of 0–2 were considered as low quality, while final scores of ≥ 3 were regarded as representing studies of high quality. Furthermore, the trials were assessed for methodological quality using the risk of bias assessment tool by the Cochrane Collaboration [18] (Figure 1).

In this systematic review, HP dietary protocols were compared with LP regimens with respect to their effects on biomarkers of obesity and obesity-associated disorders such as diabetes or cardiovascular disease. Analyses were restricted on HP as well as LP diets providing ≤ 30% of TEC in the form of fat to prevent potential bias due to variations in total fat intake. Main findings suggest no advantages or disadvantages of a higher dietary protein content. None of the dietary protocols turned out to be superior to its counterpart with regard to the biomarkers under investigation. Following primary analysis, decreases in fasting insulin were significantly more pronounced in HP diets. However, this was no longer valid after inclusion of high quality trials only in the secondary analysis. The raise in HDL-C turned out to be more pronounced in the HP group compared to the LP group following sensitivity analysis excluding studies that enrolled patients with T2D. In a previous study, HP diets exerted a 12%-increase in HDL-C under closely supervised dietary control [37]. Two meta-analyses provide evidence that higher fat intake was associated with higher levels of HDL-C when compared to low-fat diets [38, 39]. With respect to the studies included in the present systematic review, the trials by Gardner et al. [22], Dansinger et al. [26], and McAuley et al. [34] reported higher intakes of total fat at the end of their 12 months protocols (dietary records) in the HP groups as compared to the respective LP counterparts. Omitting these trials to the sensitivity analysis, changes in HDL-C turned out to be similar in both HP and LP regimen (data not shown), suggesting that HDL-C response was due to dietary fat content rather than to protein consumption.

Taken together, these results are in discrepancy with a recent meta-analysis by Santesso and co-workers [7] who reported weight loss, WC, HDL-C, TG, SBP, DBP and FI to be significantly more improved following short-and long-term HP diets as compared to LP protocols. The different findings might at least in part be explained by the fact that only long-term studies with a duration ≥ 12 months were included in the present meta-analysis. In addition, both post-intervention values as well as changes in mean differences were used as suggested by the Cochrane Collaboration [20] to avoid a standardized mean differences method, whereas Santesso et al. [7] separated between primary (change from baseline values) and secondary (final values) analyses. These results indicate that HP diets do not exert favorable effects on anthropometric measures like body weight, fat mass and waist circumference. However, in a meta-regression by Krieger et al. [11] high-protein intake turned out to be a significant predictor of fat free mass retention, thereby compensating a potential side-effect of long-term energy restriction.

Dietary protein content of the high-protein diets included in this meta-analysis varied between 30-40% of TEC, which is within the age-dependent AMDR of 5-35% for all but one RCT [31]. Via analysis of the National Health and Nutrition Examinations Survey conducted between 2003 and 2004, Fulgoni [5] concluded that the actual intake of protein in US-American adults of 1.3 g * kg body weight^-1 * d^-1 exceeds the DRI values of 0.66 g * kg body weight^-1 * d^-1. He suggested that recommendations could be adapted to 25-30% of TEC, assuming benefits of higher protein intake e.g. on regulation of body weight. Regarding biomarkers such as weight, waist circumference or fat mass, the present meta-analysis does not support this concept.

Three RCTs included in this meta-analysis investigated the effects of HP regimens on biomarkers of kidney function in patients with T2D. In all trials, HP diets did not affect renal functions assessed via measurement of serum creatinine and microalbuminurea [32, 33, 36]. Likewise, a 2-year RCT by Friedman et al. [40] reported no harmful effects of a high-protein/low carbohydrate diet on glomerular filtration rate, albuminuria, or fluid and electrolyte balance. With respect to prospective cohort studies, a systematic review by Mente et al. [41] indicated no significant correlations between animal protein sources, e.g. eggs, milk or meat on coronary heart disease (CHD), whereas vegetable protein sources like nuts were associated with a decreased risk. Findings from Greece, Sweden and the US noted an increased all-cause mortality following a HP/low carbohydrate diet based on animal sources in both women and men whereas a vegetable-based low-carbohydrate diet was associated with lower all-cause and cardiovascular disease mortality rates [42–44].

This systematic review did not consider unpublished data, and with respect to the moderate asymmetry of the Funnel plots, it cannot be excluded that publication bias such as lack of published studies with inconclusive results may have at least a moderate impact on the effect size estimates. An important limitation of dietary intervention trials is the heterogeneity of various aspects and characteristics of the study protocols. The literature chosen for the present meta-analysis varies regarding type(s) of diets used, definitions of HP and LP diets, study population (i.e. BMI, type 2 diabetics, abnormal glucose metabolism), intervention time, nutritional assessment as well as long-term follow-ups (between 1 and 2 yrs.). In addition, some studies were performed on hypocaloric terms, while others provided an isocaloric diet.

Not all of the studies gave details on the quality of their respective setup (e.g. method of randomization, follow-up protocol with reasons for withdrawal) yielding Jadad scores < 3. However, following sensitivity analyses including high quality studies only (Jadad score ≥ 3), pooled estimates of effect size were similar to those obtained with the complete set of studies. Some comparisons within the present meta-analyses were done using both post-intervention values and changes in mean difference, which is considered to be a legitimate procedure as described by the Cochrane Collaboration [20], and should not be regarded as a limitation.

In summary, the present meta-analysis investigated the long-term effects of HP vs. LP both low in fat on biomarkers predicting the outcome of obesity, cardiovascular disease and glycemic control. Since biomarkers under investigation were not affected by changes in dietary protein content, unanimous recommendation of a high-protein dietary approach is not supported by evidence. With respect to the potential risk of high-protein contents, further studies are required before dietary recommendations can be changed towards a higher percentage of daily protein consumption.