• Maximizes nutrient uptake in muscle.
• Releases and burns fat-cell content.
• Prevents the storage of fat.

Indigo-3G® acts as a "selective nutrient repartitioning agent" - meaning it maximizes nutrient uptake in muscle, releases and burns fat-cell content, and prevents storage of fat. In simple terms, Indigo-3G® repartitions nutrients so that what you eat turns into muscle and not fat.

The formula contains a self-microemulsifying delivery system of cyanidin-3 glucoside (C3G) that substantially enhances its potency, bioavailability, and stability. Most importantly, Indigo-3G® yields much higher blood levels of C3G® than any of the previous research indicated was possible, and provides the amount of C3G® required for maximum results.

New! - More Potent, Lower Cost Formula

Indigo-3G® has been and continues to be a major area of our ongoing research. Not only are we interested in understanding more about its wide-reaching and powerful effects, we're also looking for ways to increase the formula's potency (per milligram) and thereby decrease the cost to the consumer.

Improved Formula

As a reminder, Indigo-3G contains a self-microemulsifying delivery system of C3G that substantially enhances its potency, bioavailability, and stability. And recently we were able to recalibrate the delivery system to yield the same (or better) effect from 100 mg per capsule as we used to get with 125 mg per capsule. In other words, we're using "less to do more."

New Dosing Schedule

We were also able to reduce the dose schedule from three times per day to twice per day, making dosing not only more effective, but much more convenient, as well.

Auto-Ship Update

The Auto-Ship premium is switching from "bonus bottles" to a special discounted price of $97.50 per bottle. For more information about our Auto-Ship program, call 800-525-1940.

Customer Service agents are available by phone Monday through Friday, from 9 AM to 5 PM, MT.

"Everything I eat seems to turn to muscle."

When I started taking the dark-indigo capsules I weighed 221. Eight days later I had dumped fat and excess water and dropped down to 211. At first I thought I was taking a fat-burner. But after six days of adding carbs (like you suggested) I'm up to a lean 224, and I'm just as lean (if not leaner) as I was at 211!

I hate to be 'that guy' who keeps on raving about something, but I honestly would take up a second mortgage to make sure that I never run out of this stuff. It keeps getting better and better. The only downside is that I'm changing too fast and people are starting to 'talk.'"

– Christian Thibaudeau

"As a competing bodybuilder, I'm totally shocked with the effects of Indigo-3G."

I used Indigo-3G® in the four weeks between two of my bodybuilding shows, and I have to tell you that I'm totally shocked with the results. In the four weeks between shows, not only have I gotten tighter and bigger, I now have striations I've never seen before.

As a natural competitor – and almost 40! – that's simply not supposed to happen in four weeks between shows.

Not only that, I won the last contest!

– The Mighty Stu

"It's like being genetically gifted."

I'm now at the heaviest I've ever been, but I'm actually looking leaner with each passing week! The last time that I looked this lean I was on a strict diet and much lighter. I can tell a big increase in my work capacity as well. I can push my training to new levels, but my body seems to want more.

I now get what it's like to be genetically gifted.

– Barachiel

"He's F'n huge!"

I've lost 2 inches off of my waist and 2 inches off of my butt, and I'm looking a lot bigger. Even other people are commenting on the differences they're seeing in my body.

One of the powerlifters at my gym walked by while I was doing military presses and asked if he could borrow my arms for his bench press.

The next day, I was at a local festival with my daughter. Some guy from her high school saw us and texted her, "Is that your dad you're with?" She replied, "Yeah." He replied back, "He's F'n huge!"

I may not be huge relative to the rest of T-Nation, but for a 45-year-old, those comments can make your day.

– 8thMan

"I'm adding weight on every set."

After being on Indigo-3G® for several weeks, I'm getting so strong so quickly that I have to check every day to make sure that I'm putting 45's on the bar and not 35's. I'm adding more weight on every set and can't put enough weight on the bar. And I'm getting leaner, too!

I didn't expect results to be this sudden. I expected it to happen slowly over a few weeks, but I felt results within the first week. And I've never been able to lose body fat and increase strength at the same time.

People who've seen my results are commenting on the changes and asking where to get Indigo-3G.

– andrew_live

"Amazing pumps, like I'm going to explode!"

I had my neighbor come up out of the blue, squeeze my arms, and ask me about training. And she's never done that before.

The pumps I'm getting are amazing. I felt like I was going to explode. I've also noticed a great increase in work capacity, and find myself wanting to stay in the gym longer and longer to the point where I have to force myself to leave.

– ashylarryku

"I'm seeing my love handles disappearing!"

When I first started taking Indigo-3G, I was a little cautious about increasing carbs. But after seeing my love handles and lower-back fat disappearing, I'm excited to keep going with more carbs. In fact, I'm now eating as many carbs as when I was in high school.

Today at the gym I weighed 197. I haven't been under 200 in a long time. My lifts and strength are all up and energy is high. People at work have noticed something big time, too. And I had a guy at the gym ask if I was doing something different because my shirt "looked smaller."

– bbone32

"Indigo-3G® has exceeded my expectations."

I've always looked at supplements as providing a slight edge, but this one has just been fantastic. I was hoping for a small improvement, but have seen much more than that. I even get random comments from people, asking what I'm on and if it's legal.

– Bcurtis1

"I now run out of time before I run out of energy."

My wife says that I look bigger and more defined – chiseled! I'm 5 pounds heavier and look as lean as I ever have. I have so much work capacity during training that I run out of time before I run out of energy. Regarding carb tolerance, even at 300 grams per day I'm having no problems at all.

– Bigiron77

"The pumps are out of this world."

I've been on Indigo-3G® for several weeks. The other day I took off my shirt and looked like I was on a cycle of steroids.

– bodz

"I don't ever want to go a day without Indigo-3G."

As a former user, Indigo-3G is the closest thing I've found to an anabolic steroid. This is the stuff. I can't believe I am literally treating it like my "stash" of anabolics back in the day. Really, this is the ONLY supplement I've ever valued this highly.

Want to know what it feels like to be on anabolics? Using Indigo-3G® is probably the closest you can get from using a supplement. It's not exactly the same, but it's pretty close. Equally impressive is the fact that Indigo-3G® is also very healthy for you, and something you should take daily to maintain good health.

– buffd_samurai

"I believe Indigo-3G® is the best supplement I've ever used."

I've never been able to eat carbs without gaining weight and adding inches to my waist size. Now, that's no longer true. With Indigo-3G, I'm eating carbs while watching inches drop off my waist! I can see a lot more definition in my shoulders, too. My bodyweight's the same, but I'm a whole lot leaner.

I also have a lot more energy in the gym now that I'm using Indigo-3G. It's not my imagination, either – other people are taking notice. The trainer at my gym is just amazed. He can't believe that I'm able to add more exercises to my workouts. I used to run out of energy before I finished my workout; now I train until I run out of time.

Indigo-3G® has given me a whole new outlook on life.

– bwbski

"I've never seen anything work like this."

Indigo-3G® is amazing! If I could buy stock in the company, I'd put half of my life savings into it. Even my wife sees a big difference in my physique. In fact, she's complimented me more times this week than the previous 15 years combined.

– chushin

"John Romaniello commented that I look bigger and leaner, and John doesn't even know that I'm on Indigo-3G!"

With Indigo-3G, my body is now handling higher carbs and calories the way I've always wished that it could, and my muscle and strength gains are the best I've ever seen.

I couldn't be happier.

– citydawg926

"My waist is getting smaller while I'm gaining overall size."

Even someone that I see on a daily basis asked me if I'm gaining muscle. And after seeing some recent pictures, my sister said, "Holy s***, your back muscles are getting fierce!"

– corstijeir

"I've been hitting new PRs every day... while losing body fat!"

When I started taking Indigo-3G, I wanted to see my strength go up, and that has definitely happened. I'm setting PRs every workout while losing body fat. My energy is also through the roof!

– cr powerlinate

"I'm 37 and leaner now than I've ever been!"

After starting Indigo-3G, I dropped 7-8 pounds right out of the gate. I'm now leaner than I've ever been, and I'm starting to gain some appreciable muscle size. Every time I walk in to some place new, people will ask me what I'm doing to look like this. Even those who see me on a daily basis are taken aback by the changes they're seeing.

To say that I'm leaner now than I've ever been before is saying something – especially considering that I'm 37 years old. I've been training in one form or another for my entire adult life, and my current workload is the heaviest it's ever been.

Do I honestly believe that Indigo-3G® has worked as advertised?

You're damn right I do! I can eat triple the amount of carbs that I did before, and seeing my waist shrink while doing so. I feel mentally focused, far less irritable than usual, and my drive to get to the gym and train, and to continue training once I'm there, is unlike anything I've experienced before.

– Dave-G

"You have to be kidding me!"

Two weeks after starting Indigo-3G, my ex came over to drop off my son. My shirt was off and she came up to me and said, "You have to be kidding me!" She's a personal trainer and in phenomenal shape right now. She never used to compliment me when we were still married.

And one of my friends that hasn't seen me in a while saw me right after I finished a workout and said, "You're juicing!"

I love these compliments!

– emenef

"I don't think you will hear of a negative experience."

I'm seeing a big change in my whole body. People are asking, "What are you doing?" Endurance is up, libido is awesome, stamina is up.

– HawkEye0487

"I honestly feel that I can't get fat while on Indigo-3G."

So far, I gained 4 pounds and lost an inch and a half on my waist.

– Hip Scar

"As a performance enhancer, I've never taken a supplement that comes close to what I'm experiencing on Indigo-3G."

I noticed endurance and strength effects immediately, and I'm seeing body recomposition from week to week.

On vacation, I had random people coming up and asking me what I'm on. The trainer at the gym hasn't seen me since I had my body fat measured, and he couldn't believe the changes I've made. He asked me what I was on and where he could get some!

As an ex-anabolics user, I've never used a supplement that even comes close to giving the effects of anabolics... until now!

Indigo-3G® is the best supplement I've I ever used. Period!"

– Hospitaller

"I'm amazed at the progress I'm seeing."

With Indigo-3G, it's not a choice anymore between losing fat and building muscle. You can do both at the same time. Plus, my muscles look full all the time. I've never had this kind of performance improvement before.

– Itisfinished

"My workout energy is off the charts – awesome!"

Workout energy is off the charts. I'm now able to train longer and add in assistance work after my planned training session. I've never been able to do that before. As for body recomposition, I'm losing fat and gaining muscle. I've never had that happen before at the same time.

– Long Trang

"I have never experienced anything comparable to this."

I take Indigo-3G® in combination with Surge Workout Fuel, Finibars, and Anaconda, because I believe taking advanced peri-workout nutrition is the best thing you can do as a natural bodybuilder. I have never used steroids and have no idea how it feels being on them, but I just cannot imagine any more dramatic effects than with this Biotest super-combo.

Additionally, far more important to me than getting big and strong is health, and living a healthy lifestyle. That's why I love Indigo-3G® even more – it's healthy!

I'll definitely be reordering Indigo-3G, that's for sure.

– ParagonA

"The product definitely met the hype and then some!"

Even though my body weight is the same, I'm losing fat. In fact, I've lost 3 inches from my waist and had to go and buy a new belt. I'd just purchased a bunch of a new pants a few months ago and now I can't wear them because they're too loose.

– sjsosu

"As a competing bodybuilder, I've never been this lean and this heavy."

I started taking Indigo-3G® two days after competing in a bodybuilding show, and without a doubt I was able to pound the carbohydrates without gaining fat. I used to eat around 250 grams of carbs a day, but now I'm taking in close to 500 grams on hard workout days. I totally can't believe it.

– synergy93

Powerful Nutrient Repartitioning

Scientists have discovered a naturally occurring compound - called cyanidin 3-glucoside (C3G) - that has the ability to precisely control nutrients so that what you eat literally turns to muscle instead of fat.

This might seem hard to believe, but research has proven C3G's effects on insulin signaling, selective insulin sensitivity, glucose and nutrient repartitioning, as well as its direct effect on fat and muscle cells, is extremely elaborate and precise.

C3G® literally makes your body function perfectly for building muscle mass without the fear of gaining fat, even while on a bulking diet.

The body-building process is now a whole new ballgame. No more depriving yourself of the optimal nutrition for building muscle because "your body can't handle the calories and carbs" - because now it can.

Dysfunctional Nutrient Uptake

Here's an interesting fact that you might not know. The fat cell (not the muscle cell) is a major regulator of how easily you gain muscle. It's the fat cell that sends all of the molecular signals for glucose and nutrient uptake in both fat and lean tissue. This is especially important because it's the nutrient uptake (not androgens) that has the most impact on gaining muscle.

As most advanced bodybuilders know, the most anabolic hormone in the body is insulin, not testosterone, and insulin is what determines nutrient uptake. Unfortunately, insulin is anabolic in both muscle and fat. But by decreasing the effects of insulin on fat and increasing the effects of insulin on muscle, you can achieve what no one thought was possible and make the common man look like a muscle-building genetic freak.

As always, there are genetic limitations to how much muscle an individual can gain - I think we all understand that C3G® can't turn short muscle bellies into long muscle bellies. The real issue at hand, however, is not the genetic limits on overall muscle size, but specifically dysfunctional nutrient uptake. That's the real limiting factor.

The sad fact is, dysfunctional nutrient uptake prevents people from ever attaining even a fraction of the gains they could, both in terms of muscle gains and overall leanness.

"I Want to Get Big... No, I Want to Get Shredded..."

Dysfunctional nutrient uptake results in what amounts to a bodybuilding identity crisis, where you go back and forth between trying to get big and trying to get shredded - but never truly achieve either goal.

You get bigger, alright, but it's mostly fat. You tend to rationalize it until the day you see some guy in the gym with razor-sharp etched muscle who has your look - at least, the one you've decided you now want. You're sick about what you've done to yourself, and with renewed vigor you're off to shredded-land. Except, at the end of that jaunt, you haven't etched anything and you're now "skinny," which totally ticks you off and sends you back in the other direction.

This is obviously an irrational approach to achieving a lean and muscular body, but what's not so obvious is the cause of this dysfunction. It's due to a condition that's tantamount to "inflammatory fat disease."

The Cause - Inflammatory Fat Disease

Most diseases of modern living have been linked to inflammation. So it's no wonder that researchers have found that inflammatory signaling has also caused fat cells to become dysfunctional, resulting in high circulating glucose levels, out of control appetite, fat cells that love to get "fatter," and lazy muscle cells that prefer to leave most of the fuel for the fat cells to gobble up and store.

This is where C3G® comes in. The consensus among researchers is that C3G® acts primarily (through the Glut4-RBP4 system) to optimize molecular crosstalk between fat cells, which directly decreases adipocyte inflammatory signals. The overall effect causes the body to repartition nutrients so that what you eat literally turns to muscle instead of fat.

In other words, your body resists storing fat, releases and oxidizes fat-cell content (fatty acids), and preferentially pulls glucose and nutrients into muscle cells.

Here's how C3G® specifically affects fat cells and muscle cells:

Fat Cells - C3G® actually up-regulates gene expression for fat oxidation (fat burning) and down-regulates gene expression for fat storage. The oxidation of fatty acids increases further, directly through the release of adiponectin. The overall effect is that fat cells begin to shrink in size. The reduced inflammation also decreases the size of fat cells and disgorges the excess water associated with the inflammatory process (leading to a less bloated and squishy feeling to the fat under the skin).

Muscle Cells - Insulin sensitivity of muscle cells increases, thereby enhancing insulin signaling and glucose and nutrient uptake, along with enhancing glycogen synthesis, which drives glucose and nutrients preferentially into muscle.

Effective C3G® Dosing

It's simple. Find blackberries that actually contain high levels of C3G® and then eat literally 40 pounds of them per day. Too bad that's not really feasible. Besides, blackberries (and the other berries and their extracts) that contain C3G® also contain quercetin and other compounds that can either inhibit or diminish the effects of C3G.

So, even if you had a bottomless pit of a stomach and access to an unlimited supply, it's impossible to eat enough blackberries or berry extract to achieve any repartitioning effects from the C3G® they contain.

To make C3G® even workable as a super-repartitioning compound, it needs to be formulated with enhanced bioavailability and stability (to prevent oxidation). So in summary, to be highly effective, you need a pure and stable C3G® compound that's bioengineered with enhanced bioavailability.

Of course, there are a small handful of supplements on the market formulated with crude extracts that claim to contain C3G. But just as the case with fresh berries and berry extracts, none of these products contain the purified form of C3G, or anywhere close to the amount of C3G® required to be effective for nutrient repartitioning. Furthermore, none are designed to enhance the absorption or stability of C3G.

Indigo-3G® is the only C3G® formula in the world with the potency, bioavailability, and stability required for effective, maximum results.

Results - What to Expect

Indigo-3G® could very well change your life from this point forward - change the way you handle carbs, change the way you don't store body fat, and change the way you gain muscle. But you better get an action plan before you start the program, or you very well could miss out on the very best physique gains of your life.

Here's a fact: dramatic physique changes can't come overnight. Heck, they can't even come quickly, especially if you want them to last. We work with the gifted elite, and even these physique champions have to work brutally hard over time (just like you and me) to make impressive gains.

So don't expect to take Indigo-3G® tonight and wake up tomorrow morning transformed into the new you. You'll eventually get that new-you image; it's just going to take some honest hard work and a little time to get there. So be patient and watch the process become a whole lot easier and consistent with Indigo-3G®.

Your current physiology and body composition will determine what types of results you experience. But regardless of your condition, everyone should take Indigo-3G® for two weeks before evaluating the effects. That's the "break-in" period (the first two weeks). Beyond that is when things begin to really take shape.

So far, all of our subjects have experienced significant results in the areas of overall improved body composition, pronounced muscle fullness, increased rate of strength gains, universal carb tolerance, and being resistant to gaining fat. Many users have gone so far as to say, "I now know what it's like to be one of the lucky ones who can eat 'normally' and not get fat. I no longer have to worry about eating 'perfectly' to see my abs."

To make sure you experience the greatest effects from Indigo-3G, here are the important things to keep in mind:

Enhanced Carb Uptake

You can't repartition something you don't consume. So, if you don't consume ample supplies of nutrients, especially extra carbs in peri-workout nutrition, you'll greatly diminish muscle mass gains.

In contrast, low-carb diets make your muscles feel and appear flat. This "flatness" is due to low intramuscular pressure, which is associated with a catabolic (muscle wasting) state. Keeping muscle bellies nice and full, by including ample amounts of carbohydrates, means that you're not losing muscle mass and well on the way to building some.

Increased Muscle Gains and Fat Loss

Most users will begin noticing changes in body composition within the first week or two. Some have even reported substantial muscle gains and fat loss during that same period. As a general rule, those who have above average muscle mass and train often (5 or 6 days a week) will experience a more rapid rate of muscle mass gains and fat loss.

Initially, you might not see a change in overall body weight, but notice that you're getting leaner and more muscular. We've observed this weight homeostasis effect when doing Dexa scans on subjects, so don't let the number on the scales negatively influence your judgment.

Larger Muscle Pumps

Within a few days, muscle pumps should come much more quickly and easily, and be noticeably larger (even to others in the gym), sometimes to the point of substantially affecting movement. The pump should also last longer after the workout is completed. Even at rest, the muscles of Indigo-3G® users can feel and appear fuller and semi-pumped, especially when taking in ample amounts of proper quality nutrients.

Faster Strength Gains

Strength increases should come faster and more often. In fact, some reported having to recheck the plates on the bar after a set, thinking they forgot to add weight between sets.

Enhanced Work Capacity and Recovery

Due to enhanced glucose management, almost all users notice what many describe as "limitless energy to do more in the gym." And they also report improved recovery rates. As an extreme example of this, Christian Thibaudeau recently had a monster training day where he performed a total of 258 sets (this is not a typo) spread over 3 sessions, followed by another two workouts the very next day. That's FIVE hardcore-to-the-bone workouts over two days, with no recovery problems. Granted, he included our most advanced peri-workout nutrition protocol in every training session. Even so, Indigo-3G® potentiated the overall effects to a whole new level.

The First Step

Before you get start taking Indigo-3G, you really need to take a hard look at your current body and physique condition, and be totally honest with your assessment. To do that, I want you to strip down to your skivvies in front of the mirror - do it! Be brutally objective about what you see. And don't forget to check out that backside of yours. This might be the first time in a long time that many of you have looked back there, and it could possibly be a bit, shall I say, traumatizing.

When you've finished with the body-scan, no matter what you've seen, again, be totally honest with yourself and embrace whatever physique challenges lie between where you are now and where you ultimately want to be.

Now, let's get to work and make your goals a reality.

ABSTRACTS

Cyanidin 3-glucoside ameliorates hyperglycemia and insulin sensitivity due to downregulation of retinol binding protein 4 expression in diabetic mice.

Abstract

Adipocyte dysfunction is strongly associated with the development of obesity and insulin resistance. It is accepted that the regulation of adipocytokine expression is one of the most important targets for the prevention of obesity and improvement of insulin sensitivity. In this study, we have demonstrated that anthocyanin (cyanidin 3-glucoside; C3G) which is a pigment widespread in the plant kingdom, ameliorates hyperglycemia and insulin sensitivity due to the reduction of retinol binding protein 4 (RBP4) expression in type 2 diabetic mice. KK-A(y) mice were fed control or control +0.2% of a C3G® diet for 5 weeks. Dietary C3G® significantly reduced blood glucose concentration and enhanced insulin sensitivity. The adiponectin and its receptors expression were not responsible for this amelioration. C3G® significantly upregulated the glucose transporter 4 (Glut4) and downregulated RBP4 in the white adipose tissue, which is accompanied by downregulation of the inflammatory adipocytokines (monocyte chemoattractant protein-1 and tumor necrosis factor-alpha) in the white adipose tissue of the C3G® group. These findings indicate that C3G® has significant potency in an anti-diabetic effect through the regulation of Glut4-RBP4 system and the related inflammatory adipocytokines.

Biochem Pharmacol. 2007 Dec 3;74(11):1619-27. Epub 2007 Aug 10.

Anthocyanin enhances adipocytokine secretion and adipocyte-specific gene expression in isolated rat adipocytes.

Abstract

Adipocyte dysfunction is strongly associated with the development of obesity and insulin resistance. It is accepted that the regulation of adipocytokine secretion or the adipocyte-specific gene expression is one of the most important targets for the prevention of obesity and amelioration of insulin sensitivity. In this study, we demonstrated that anthocyanins (cyanidin or cyanidin 3-glucoside) have the potency of a unique pharmacological function in isolated rat adipocytes. Treated adipocytes with anthocyanins enhanced adipocytokine (adiponectin and leptin) secretion and up-regulated the adipocyte specific gene expression without activation of PPARgamma in isolated rat adipocytes. The gene expression of adiponectin was also up-regulated in white adipose tissue in mice fed an anthocyanin supplemented diet. As one of the possible mechanisms, AMP-activated protein kinase activation would be associated with these changes, nevertheless, the AMP:ATP ratio was significantly decreased by administration of the anthocyanins. These data suggest that anthocyanins have a potency of unique therapeutic advantage and also have important implications for preventing obesity and diabetes.

Biochem Biophys Res Commun. 2004 Mar 26;316(1):149-57.

Cyanidin 3-glucoside protects 3T3-L1 adipocytes against H2O2- or TNF-alpha-induced insulin resistance by inhibiting c-Jun NH2-terminal kinase activation.

Abstract

Anthocyanins are naturally occurring plant pigments and exhibit an array of pharmacological properties. Our previous study showed that black rice pigment extract rich in anthocyanin prevents and ameliorates high-fructose-induced insulin resistance in rats. In present study, cyanidin 3-glucoside (Cy-3-G), a typical anthocyanin most abundant in black rice was used to examine its protective effect on insulin sensitivity in 3T3-L1 adipocytes exposed to H(2)O(2) (generated by adding glucose oxidase to the medium) or tumor necrosis factor alpha (TNF-alpha). Twelve-hour exposure of 3T3-L1 adipocytes to H(2)O(2) or TNF-alpha resulted in the increase of c-Jun NH(2)-terminal kinase (JNK) activation and insulin receptor substrate 1 (IRS1) serine 307 phosphorylation, concomitantly with the decrease in insulin-stimulated IRS1 tyrosine phosphorylation and cellular glucose uptake. Blocking JNK expression using RNA interference efficiently prevented the H(2)O(2)- or TNF-alpha-induced defects in insulin action. Pretreatment of cells with Cy-3-G reduced the intracellular production of reactive oxygen species, the activation of JNK, and attenuated H(2)O(2)- or TNF-alpha-induced insulin resistance in a dose-dependent manner. In parallel, N-acetyl-cysteine, an antioxidant compound, did not exhibit an attenuation of TNF-alpha-induced insulin resistance. Taken together, these results indicated that Cy-3-G exerts a protective role against H(2)O(2)- or TNF-alpha-induced insulin resistance in 3T3-L1 adipocytes by inhibiting the JNK signal pathway.

Biochem Pharmacol. 2008 Mar 15;75(6):1393-401. Epub 2007 Dec 3.

Microarray profiling of gene expression in human adipocytes in response to anthocyanins.

Abstract

Adipocyte dysfunction is strongly associated with the development of obesity and insulin resistance. It is accepted that the regulation of adipocytokine secretion or the adipocyte specific gene expression is one of the most important targets for the prevention of obesity and amelioration of insulin sensitivity. Recently, we demonstrated that anthocyanins, which are pigments widespread in the plant kingdom, have the potency of anti-obesity in mice and the enhancement adipocytokine secretion and its gene expression in adipocytes. In this study, we have shown the gene expression profile in human adipocytes treated with anthocyanins (cyanidin 3-glucoside; C3G® or cyanidin; Cy). The human adipocytes were treated with 100 microM C3G, Cy or vehicle for 24 h. The total RNA from the adipocytes was isolated and carried out GeneChip microarray analysis. Based on the gene expression profile, we demonstrated the significant changes of adipocytokine expression (up-regulation of adiponectin and down-regulation of plasminogen activator inhibitor-1 and interleukin-6). Some of lipid metabolism related genes (uncoupling protein2, acylCoA oxidase1 and perilipin) also significantly induced in both common the C3G® or Cy treatment groups. These studies have provided an overview of the gene expression profiles in human adipocytes treated with anthocyanins and demonstrated that anthocyanins can regulate adipocytokine gene expression to ameliorate adipocyte function related with obesity and diabetes that merit further investigation.

Biochem Pharmacol. 2006 Apr 14;71(8):1184-97. Epub 2006 Feb 17.

Gene expression profile of isolated rat adipocytes treated with anthocyanins.

Abstract

Adipocyte dysfunction is strongly associated with the development of obesity and insulin resistance. It is accepted that the regulation of adipocytokine secretion or the adipocyte specific gene expression is one of the most important targets for the prevention of obesity and amelioration of insulin sensitivity. Recently, we demonstrated that anthocyanins, which are pigments widespread in the plant kingdom, have the potency of anti-obesity in mice and the enhancement adipocytokine secretion and adipocyte gene expression in adipocytes. In this study, we have shown for the first time the gene expression profile in isolated rat adipocytes treated with anthocyanins (cyanidin 3-glucoside; C3G® or cyanidin; Cy). The rat adipocytes were treated with 100 muM C3G, Cy or vehicle for 24 h. The total RNA from the adipocytes was isolated and carried out GeneChip microarray analysis. A total of 633 or 427 genes was up-regulated (>1.5-fold) by the treatment of adipocytes with C3G® or Cy, respectively. The up-regulated genes include lipid metabolism and signal transduction-related genes, however, the altered genes were partly different between the C3G- and Cy-treated groups. Based on the gene expression profile, we demonstrated the up-regulation of hormone sensitive lipase and enhancement of the lipolytic activity by the treatment of adipocytes with C3G® or Cy. These data have provided an overview of the gene expression profiles in adipocytes treated with anthocyanins and identified new responsive genes with potentially important functions in adipocytes related with obesity and diabetes that merit further investigation.

Biochim Biophys Acta. 2005 Apr 15;1733(2-3):137-47. Epub 2005 Jan 12.

Hypoglycemic activity of a novel anthocyanin-rich formulation from lowbush blueberry, Vaccinium angustifolium Aiton.

Abstract

Blueberry fruits are known as a rich source of anthocyanin components. In this study we demonstrate that anthocyanins from blueberry have the potency to alleviate symptoms of hyperglycemia in diabetic C57b1/6J mice. The anti-diabetic activity of different anthocyanin-related extracts was evaluated using the pharmaceutically acceptable self-microemulsifying drug delivery system: Labrasol. Treatment by gavage (500 mg/kg body wt) with a phenolic-rich extract and an anthocyanin-enriched fraction formulated with Labrasol lowered elevated blood glucose levels by 33 and 51%, respectively. The hypoglycemic activities of these formulae were comparable to that of the known anti-diabetic drug metformin (27% at 300 mg/kg). The extracts were not significantly hypoglycemic when administered without Labrasol, demonstrating its bio-enhancing effect, most likely due to increasing the bioavailability of the administered preparations. The phenolic-rich extract contained 287.0+/-9.7 mg/g anthocyanins, while the anthocyanin-enriched fraction contained 595+/-20.0 mg/g (cyanidin-3-glucoside equivalents), as measured by HPLC and pH differential analysis methods. The greater hypoglycemic activity of the anthocyanin-enriched fraction compared to the initial phenolic-rich extract suggested that the activity was due to the anthocyanin components. Treatment by gavage (300 mg/kg) with the pure anthocyanins, delphinidin-3-O-glucoside and malvidin-3-O-glucoside, formulated with Labrasol, showed that malvidin-3-O-glucoside was significantly hypoglycemic while delphinidin-3-O-glucoside was not.

Phytomedicine. 2009 May;16(5):406-15. Epub 2009 Mar 20.

Dietary cyanidin 3-O-beta-D-glucoside-rich purple corn color prevents obesity and ameliorates hyperglycemia in mice.

Abstract

Anthocyanins, which are used as a food coloring, are widely distributed in human diets, suggesting that we ingest large amounts of anthocyanins from plant-based foods. Mice were fed control, cyanidin 3-glucoside-rich purple corn color (PCC), high fat (HF) or HF + PCC diet for 12 wk. Dietary PCC significantly suppressed the HF diet-induced increase in body weight gain, and white and brown adipose tissue weights. Feeding the HF diet markedly induced hypertrophy of the adipocytes in the epididymal white adipose tissue compared with the control group. In contrast, the induction did not occur in the HF + PCC group. The HF diet induced hyperglycemia, hyperinsulinemia and hyperleptinemia. These perturbations were completely normalized in rats fed HF + PCC. An increase in the tumor necrosis factor (TNF)-alpha mRNA level occurred in the HF group and was normalized by dietary PCC. These results suggest that dietary PCC may ameliorate HF diet-induced insulin resistance in mice. PCC suppressed the mRNA levels of enzymes involved in fatty acid and triacylglycerol synthesis and lowered the sterol regulatory element binding protein-1 mRNA level in white adipose tissue. These down-regulations may contribute to triacylglycerol accumulation in white adipose tissue. Our findings provide a biochemical and nutritional basis for the use of PCC or anthocyanins as a functional food factor that may have benefits for the prevention of obesity and diabetes.

J Nutr. 2003 Jul;133(7):2125-30.

Cyanidin 3-glucoside protects 3T3-L1 adipocytes against H2O2- or TNF-alpha-induced insulin resistance by inhibiting c-Jun NH2-terminal kinase activation.

Abstract

Anthocyanins are naturally occurring plant pigments and exhibit an array of pharmacological properties. Our previous study showed that black rice pigment extract rich in anthocyanin prevents and ameliorates high-fructose-induced insulin resistance in rats. In present study, cyanidin 3-glucoside (Cy-3-G), a typical anthocyanin most abundant in black rice was used to examine its protective effect on insulin sensitivity in 3T3-L1 adipocytes exposed to H(2)O(2) (generated by adding glucose oxidase to the medium) or tumor necrosis factor alpha (TNF-alpha). Twelve-hour exposure of 3T3-L1 adipocytes to H(2)O(2) or TNF-alpha resulted in the increase of c-Jun NH(2)-terminal kinase (JNK) activation and insulin receptor substrate 1 (IRS1) serine 307 phosphorylation, concomitantly with the decrease in insulin-stimulated IRS1 tyrosine phosphorylation and cellular glucose uptake. Blocking JNK expression using RNA interference efficiently prevented the H(2)O(2)- or TNF-alpha-induced defects in insulin action. Pretreatment of cells with Cy-3-G reduced the intracellular production of reactive oxygen species, the activation of JNK, and attenuated H(2)O(2)- or TNF-alpha-induced insulin resistance in a dose-dependent manner. In parallel, N-acetyl-cysteine, an antioxidant compound, did not exhibit an attenuation of TNF-alpha-induced insulin resistance. Taken together, these results indicated that Cy-3-G exerts a protective role against H(2)O(2)- or TNF-alpha-induced insulin resistance in 3T3-L1 adipocytes by inhibiting the JNK signal pathway.

Biochem Pharmacol. 2008 Mar 15;75(6):1393-401. Epub 2007 Dec 3.