Bergamot Juice and Visceral Fat: Oil, Water, and Gut Microbiota

by Anthony L. Almada, MSc, FISSN

THEATRICAL ROOTS, UNIQUE FRUITS 

Claude Debussy’s signature piece, Clair de Lune (Moonlight), first published in 1905, is derived from the four part solo piano suite, Suite Bergamasque. Bergamasque refers (in part) to a 16th century form of Italian theater called Commedia dell’Arte (comedy of the professional artists), which was later performed throughout Europe and most keenly embraced in France. Bergamasque is a French derivation of the Italian bergamasco, which included works of literature, poetry, music, and dance, attributed to a small northern city in the Lombardy region named Bergamo, resting south of the Italian Alps. 

Over 1,100 km to the southeast is the Calabrian region of Italy, the front of “the boot” of the southernmost part of the Italian mainland. Here resides the epicenter of global production of Citrus bergamia Risso et Poiteau, AKA bergamot. Bergamot fruit is large, yellow, and round. This common name does not derive from the city of Bergamo, as is commonly believed. Rather, it may originate from Greece, Turkey, or Spain, each bearing cities or descriptors in the native tongue that serve as roots for bergamot or bergamia.

Bergamot’s peel-derived oil and essence served as the inspiration for the creation of the first ever Eau de Cologne, invented in the early 1700s by transplanted Italian national Johann Maria Farina, at his brother’s company in Cologne, Germany. 

A label of original Eau de Cologne [circa 1868], invented by Johann Maria Farina. To this day the original Eau de Cologne is produced from Bergamot extract. Image: DPMA

Note: The brand name associated to this fragrant invention was rabidly duplicated by numerous competitors for over 150 years yet in 1876 the name and address became one of the first registrations of intellectual property in the newly formed German Patent and Trademark Office (DPMA), as a trademark.

OIL & WATER IMMISCIBILITY

The emergence of cologne and perfume as a global industry had bergamot peel-derived oil at one of its axes. Essence of Bergamot has also entered into the hand sanitizer world, being used as a more pleasant aroma. However, as has been seen in innumerable plant-derived industries that focus upon a certain fraction or part of the plant, a bergamot waste stream emerged. The extra-peel remnants, the fleshy part and its rather bitter juice, were deemed to be of little to negative value (an attendant cost for disposal)…until the waste stream got science.

Researchers from the University of Messina (the northeastern most part of Sicily, and a 2.5 hour southwestern drive and ferry ride from Calabria) wished to subject the folklorical use of Bergamot juice (for its claimed blood lipid-lowering properties) to the rigors of a preclinical study. Adult rats that were fed either a “normal” diet or a hyperlipidemic diet (2% cholesterol, 20% coconut oil, 44% sucrose) for 30 days. Two groups of rats were fed the latter diet, one receiving hand squeezed Calabrian Bergamot juice (CBJ) from three cultivars (1 ml/day), the other received no placebo or “sham” drink (thus the research team was unblinded as to which group received CBJ). In their 2007 paper in an American Chemical Society journal they noted that the CBJ-treated rats showed a dramatic shift in the diet-induced dyslipidemia, and a marked reduction of the fatty infiltration seen in the livers from rats that received the fatty-sweet diet yet no CBJ. This study ushered in a continuum of clinical studies.

BERGAMOT BEGETS BETTER BLOOD LIPIDS

A systematic review of clinical studies (published just over one year ago) by Spanish researchers, assessing the effects of Bergamot derivatives, revealed the emerging body of evidence in relation to the management of dyslipidemia. Twelve clinical studies met the authors’ selection criteria yet only two were randomized, double blind, and placebo controlled. Of these two only one used a Bergamot extract as the sole intervention; the other study, errantly entitled with “…Citrus bergamia extracts-based formulation…” (500 mg/day) was actually a “plant sterol esters and orange oil”-based (820 mg/day) formulation, along with very high potency B vitamins and a modest amount of vitamin C. The authors’ conclusion from this review included this portentous statement [not edited for grammar]:

“Promising findings give away from an alternative therapeutic option in dyslipidemia management with bergamot supplementation, especially in patients with moderate hypercholesterolemia, low cardiovascular risk or intolerant to traditional pharmacological treatment. However, crucial issues are still remained to be clarified before a solid recommendation on this nutraceutical could be performed. These questions include the form in which most efficiently bergamot should be provided (BPF, certain flavonoids or whole bergamot juice extraction) and the optimal dose and its safety.”

BERGAMOT FLAVONOIDS 2.0

Since the aforementioned 2019 review a short-term clinical trial was conducted with a novel Bergamot extract formulation, among persons with dyslipidemia and type 2 diabetes (T2D). The Bergamot-derived polyphenol fraction (BPF) utilized was standardized, in part, to three flavonones: neoeriocitrin, naringin, and neohesperidin. As has been observed with other citrus flavonoids (e.g. quercetin), oral bioavailability can be highly variable between persons, with up to ≈ 33% being non-responders i.e. displaying no significant increase in blood concentrations of a specific flavonoid and its metabolites. (Note: this has also been seen with drug-form cannabidiol (CBD); no rigorously controlled bioavailability studies have been conducted with consumer-spec, CBD-centric hemp extracts).

The BPF (derived from Calabrian Bergamot fruit juice) used in this study was formulated with the innovative food grade delivery system Phytosome? (BPF Phytosome?). This rational formulation strategy was developed and patented by Indena SpA in the latter half of the 1980s and leverages plant-derived phospholipids (from non-GM-derived sunflower) to form uniform complexes with water insoluble constituents e.g. flavonoids, rendering them dramatically more water soluble and, presumably, more bioavailable. 

 The BPF (derived from Calabrian Bergamot fruit juice) used in this study was formulated with the innovative food grade delivery system Phytosome? (BPF Phytosome?). This rational formulation strategy was developed and patented by Indena SpA in the latter half of the 1980s and leverages plant-derived phospholipids (from non-GM-derived sunflower) to form uniform complexes with water insoluble constituents e.g. flavonoids, rendering them dramatically more water soluble and, presumably, more bioavailable. 

This pilot study integrated modified bioavailability and functional outcomes into a 30 day intervention period. Three groups of twenty subjects (all with T2D and dyslipidemia; equally balanced among genders) were randomized to one of the three following treatments:

? Placebo tablets

? BPF tablets (650 mg; standardized to 40% BPF) 

? BPF Phytosome (Vazguard?) tablets (500 mg, providing 200 mg of BPF or ≈ 31% of the BPF tablets dosage, the balance being the Phytosome? formulation) 

Subjects were asked to take one tablet before meals, twice daily.

Both BPF interventions were associated with significant (relative to placebo) and favorable changes in circulating total and LDL cholesterol, and triglycerides, unattended by changes in HDL cholesterol. Additionally, significant shifts in lipoprotein particle size (via NMR) for HDL and IDL manifested, suggestive of a cardio-/vaso-protective outcome. Fasting glucose was significantly reduced in the two BPF groups, relative to placebo. Unfortunately, fasting blood insulin was not measured, to perform surrogate measures of whole body insulin resistance e.g. HOMA-IR or QUICKI. 

Pharmacokinetic measures of blood concentrations of naringin, naringenin and naringenin glucuronide were assessed in the subjects after taking placebo, BPF, and Vazguard, on day 0 and after 30 days of supplementation. PK values appeared similar between the two BPF treatment groups, for naringin and naringenin (naringin + a linked carbohydrate AKA glycoside), while naringenin glucuronide values were modestly higher for the unmodified BBF. This latter value suggests that enterohepatic transformation of naringenin was altered by the Phytosome? formulation. Most distinctively, the quantitatively similar pharmacokinetic responses of Vazguard? to the unmodified BPF, in light of the nearly two-thirds lesser concentration of BPF/dose, reveals an approximately 2.5 times higher blood naringin dose response, on a milligram to milligram basis of BPF.

Plasma naringin concentration response after oral supplementation with a single dose of BPF (650 mg), or BPF (200 mg, in a Phytosome?: Vazguard?). From: Mollace et al., 2019.

The pharmacodynamic (blood lipids and glucose) and pharmacokinetic responses indicate the two treatments to be bioequivalent, despite the amount of BPF ingested from Vazguard? being over three times less than the standard BPF formulation. Alternatively stated, in this study among persons with T2D and dyslipidemia, Vazguard? achieved the same outcomes as BPF, at doses less than one-third of BPF.

Vazguard? to ViscGuard?

One of the most common (and inexpensive) outcome measures to collect in a clinical trial is body mass; one of the most expensive, non-invasive outcome measures is visceral fat mass, as it usually requires a magnetic resonance imaging (MRI), computerized tomography (CT), or dual energy X-ray absorptiometry (DEXA) scan, all employing expensive medical equipment and relatively longer analysis time, performed by a trained technician. A compelling body of evidence indicates that excess amounts of visceral fat are strongly associated with several metabolic dysregulatory manifestations; the metabolic aberrations directly mediated by visceral fat include hypertriglyceridemia, increases in C-reactive protein, and (possibly) insulin resistance of the liver. Previous studies assessing the impact of Bergamot juice fractions on cardiovascular and metabolic health profiles have either failed to measure changes in body mass, especially visceral fat, or have done so and observed no significant changes. 

A recent clinical trial conducted by researchers at the University of Pavia, Italy, in collaboration with research scientists from Indena SpA, marks the first intervention with an extract of Bergamot that assessed visceral fat mass shifts in response to chronic supplementation. Vazguard? is the first and only Bergamot extract shown to distinctively reduce visceral fat mass in humans.

Sixty-four overweight to obese, sedentary subjects (36 females; 56.2%) presenting with mild hyperlipidemia, mild insulin resistance, lacking a history of cardiovascular disease, and between the ages of 18-60, were recruited. Subjects were then randomized to receive placebo or Vazguard? tablets, the latter delivering 200 mg of BPF/tablet. They were asked to take one tablet at breakfast and one at dinner, for twelve weeks. The diet prescribed to the subjects, and reinforced by a dietitian at study entry, aimed for 28% of energy from fat (cholesterol <200 mg), 57% of energy from carbohydrates (10% from simple carbohydrates), with 20–25 g of bran and 15% of energy from protein. 

Body composition and blood parameters were assessed at 30, 60, and 90 days of supplementation. Body composition was assessed via Dual-Energy X-ray Absorptiometry (DXA; the paragon technique of assessing body composition in clinical research), with regional fat calculated to assess visceral and lower body fat mass.  The primary outcome measure was visceral fat and the study was powered to this, requiring a minimum of 60 subjects. Fat-free mass (FFM; inclusive of bone mass) was also quantified. Blood variables included these secondary outcome measures:

? Total, LDL, HDL, and VLDL cholesterol; triglycerides

? Fasting glucose and insulin; HOMA-IR (Homeostatic Model Assessment of [whole body] Insulin Resistance, a score derived from this formula: (Fasting insulin) X (Fasting glucose) / 405

? Apo A1, ApoB, and ApoA1/HDL ratio (Apo is short for Apolipoprotein; Apo A1 and B are the principal protein transporters of HDL and non-HDL particles, respectively, and they may offer additional precision in predicting cardiovascular disease risk)

 ? AST, ALT, and gGT (These are circulating enzymes that are indices of liver function or injury, with injury very frequently associated with significant increases)

After 30 days of Vazguard supplementation (and a dietitian-guided diet) significant reductions in DXA-determined visceral fat were observed, relative to placebo. These were maintained through the end of the study. This equated, as an average across the Vazguard group, to 151 grams or ≈ one-third of a pound, which endured through the duration of the twelve weeks. The placebo group showed a 35-gram decline by the end of the study. Surprisingly, although FFM remained stable throughout the study within the Vazguard group, the placebo group demonstrated a 2.35 kg (5.2 pound) decline by study end—the differences between the two groups were not significant. This may have been due to several of the placebo subjects experiencing an even larger reduction in FFM relative to the placebo group average. Whole body mass changes did not differ between groups, with both groups displaying modest declines over the course of the study.

Three day (two weekdays and one weekend day—dietary patterns shift on weekends) weighed food records were recorded at the beginning and end of the study. 

Significant and favorable improvements were observed in the Vazguard? group for these outcomes, relative to placebo, from the beginning to the end of the study:

? ApoA1 (increased)

? ApoB/ApoA1 ratio (decreased)

? Total cholesterol/HDL ratio (decreased)

? LDL/HDL ratio (decreased)

No adverse effects were reported in either group. Additionally, the liver enzymes profiles remained unchanged throughout the study, in both groups. The absence of any photochemical skin reactions among the subjects randomized to receive Vazguard? may be attributed to the presence of ultra trace concentrations of furocoumarins (bergamottin, bergaptene) in Vazguard?, relative to the un-extracted, whole juice.

Vazguard?, a precision extract of Bergamot, integrated with Phytosome? technology, is the first and only Bergamot extract shown to distinctively reduce visceral fat mass in humans. Additionally, the findings of favorable blood lipoprotein/apolipoprotein shifts suggest a cardioprotective effect. It is of note that although the beginning and end of study dietary intake (total energy intake [calories], macronutrients) data were not reported in the paper, communications with the authors revealed that no significant between group differences were observed, suggesting that Vazguard? may alter adipose/lipid storage and utilization kinetics, and possible de novo lipogenesis (new synthesis of lipids). This warrants exploration in future studies.

A Possible Gut Microbiota Link?

Recent in vitro studies by scientists at Indena suggests that Vazguard? may induce shifts in alpha diversity of the human fecal microbiota, which may be related to alterations in whole body lipid metabolism and lipid storage. One of the genera altered in these experiments was Blautia species.  This genus has a demonstrated ability to transform certain flavonoids into novel compounds. It is of note to mention that fecal Blautia species abundance has been shown to be inversely related to visceral fat in cross-sectional studies in humans, both age- and gender-independent.

Additional clinical studies exploring the influence of Bergamot extracts and Vazguard? on the colonic and fecal microbiota, and on adipogenesis, excess lipid storage, and insulin sensitivity and lipid metabolism, will open new vistas into blood and adipose tissue lipidomics.

Mr. Almada is a paid consultant to Indena SpA, innovators and marketers of Vazguard?.