I've been taking this stuff for over 3 years now, and it hasn't killed me yet.
In a previous peer-reviewed study commissioned by the Company that makes Protandim it was shown that after 120 days of supplementing with Protandim, erythrocyte SOD increased by 30% ( p < 0.01) and catalase by 54% ( p < 0.002). You can read this original study here,
http://www.protandim...erJan152006.pdf
Now there's a new more independent study. In this peer-reviewed study about Protandim published 31 October 2008, in the journal Free Radical Biology and Medicine they found that Protandim increased glutathione by 300%. Dr. Joe McCord, a co-author of the study, commented, “The results of this study may change how we view aging and the factors that impact healthy aging. Notably, glutathione, which is of particular interest to many disease researchers, was increased more than 300 percent with Protandim. You can read the 39 page study here,
http://www.protandim...ndim-study1.pdf
ACKNOWLEDGEMENTS: This work was supported by grants from Juvenile diabetes Research Foundation (5-2005-1104, to S.P.), American Diabetes Association (1-06-JF-40, to S.P.) and LifeVantage Corp. (to S.P. and J.M.M.). We thank the support provided by Microscopy Core facility at Denver VA Medical Center.
Here's 4 pages of the new study.
Synergistic induction of Heme oxygenase-1
Discussion:
The nutraceutical supplement Protandim has been shown to reduce the age dependent increase in the accumulation of circulating products of lipid peroxidation in healthy subjects [12]. In this study, we demonstrate that the phytochemical ingredients present in Protandim exert synergy in inducing heme oxygenase-1 (HO-1), a cytoprotective phase 2 enzyme, in cultured MIN6 and SK-N-MC cells. The effect of Protandim was significantly more than the sum of the effects of individual components. Omission of any one of the ingredients including those that did not have any independent effect reduced the activity of Protandim significantly. Curcumin was found to be the most active component of this supplement with respect to HO-1 induction. The
induction by Protandim involved the presence of ARE sites in HO-1 promoter and the nuclear localization of the transcription factor Nrf2. Involvement of multiple signaling pathways mediated by PI 3-kinase/Akt, p38 MAPK and PKCd appears to be the probable mechanism for the synergy among the components of Protandim. Furthermore, Protandim elevated the glutathione content of cells, a marker for the cellular defense against oxidative stress. This study suggests that induction of antioxidant enzymes by a combination phytochemicals at low doses is an efficient and safe approach to reduce oxidative stress in chronic diseases.
In response to oxidative stress and xenobiotic insult, phase 2 enzymes are induced as part of the cellular defense. The electrophiles generated by phase 1 enzymes (such as cytochrome P450s) are scavenged by phase 2 enzymes including HO-1, g–glutamylcysteine ligase, glutathione S-transferase and NAD(P)H:quinone oxidoreductase [27]. These enzymes contain ARE sites in their promoter region and are induced by the transcription factor Nrf2. Because Protandim induces HO-1 through Nrf2, we can anticipate that it could induce other phase 2 enzymes as well although the degree of induction is likely to vary depending on the number of ARE sites in the promoter region. Coordinated induction of a family of enzymes with antioxidant and
detoxification properties is likely to have therapeutic value. HO-1, in particular has emerged as an important mediator of cellular defense against wide ranging tissue injuries and has been suggested to be a therapeutic target in various disease models [15, 28, 29]. In addition to its antioxidant action by degradation of heme, HO-1 also exerts beneficial effects through the byproducts of heme degradation, namely CO and biliverdin [15]. The cytoprotective actions of HO-1 in pancreatic b cells which are known to express antioxidant enzymes at low levels have been well documented. For example, induction of HO-1 in mouse islets by protoporphyrin improves islet function and survival after transplantation [30]. HO-1 upregulation leads to protection of b cells from cytokines and Fas [30-32]. Overexpression of HO-1 in rat islets reduces lymphocyte infiltration in
the transplanted islets suggesting anti-inflammatory effects [33].
In this study, we used a neuroblastoma cell line (SK-N-MC) and a mouse b cell line (MIN6) to test the induction of HO-1 by Protandim. Our main objective was to determine if Protandim could be used as an antioxidant supplement in the context of neurodegenerative diseases and in diabetes. The brain is vulnerable to oxidative stress because of its high glucose-driven metabolic rate, high polyunsaturated fatty acid content, and high enzymatically active transition metal content [34]. The brain (2-3% of body weight) consumes 20% of the oxygen supply to the body, and 1-2% of the total
oxygen consumed will form reactive oxygen species (ROS). Oxidative stress and accumulation of free radical-induced damage is an important feature of aging. Markers of oxidative stress are found in aged rats, especially in those with impaired spatial learning [35]. Lipid peroxidation, DNA oxidation products and markers of protein oxidation accumulate in AD brains as a result of oxidative stress [36-39]. Tg2576 mice, a mouse model for AD treated with a combination antioxidant/anti-inflammatory agents have decreased protein carbonyls and decreased Ab levels [40], suggesting that
oxidative stress precedes AD pathology.
The pancreatic b cells are particularly vulnerable to oxidative stress-induced injury due to low level expression of antioxidant enzymes [1, 19]. Oxidative stress is known to play an important role in b cell dysfunction and loss in both types of diabetes. In type 1 diabetes, the cytokines released from immune cells that infiltrate islets generate free radicals including nitric oxide [41]. In type 2 diabetes, although insulin resistance is considered to be the primary defect, glucotoxicity resulting from chronic hyperglycemia is to known to cause b cell dysfunction and loss through generation of free radicals [42]. Thus antioxidant therapy is likely to be beneficial in improving b cell mass in diabetes. Furthermore, oxidative stress plays an important role in the loss of b cells in transplanted islets [43]. Islets are subjected to oxidative stress during isolation, storage and after transplantation. Overexpression of antioxidant enzymes in islets ex vivo has been shown to improve their function after transplantation [44, 45]. The biological actions of curcumin, Silymarin and EGCG have been extensively studied. Very limited information is available regarding the other two ingredients namely ashwagandha and Bacopa. Both are used in Ayurvedic medicine and studies have demonstrated their beneficial effects. Alcoholic extract of ashwagandha when
administered in rats exerts neuroprotective effects against 6-hydroxydopamine induced oxidative stress [46]. The markers of oxidative stress were improved by Ashwagandha. Several studies have demonstrated the antioxidant effects of extract of Bacopa monniera in vivo [47-49]. The active glycosides from this herb have been isolated and characterized [50].
Although curcumin showed the maximum effects in the induction of HO-1, it will be difficult to predict the same with other antioxidant enzymes especially the ones not regulated by Nrf2. For example, SOD and catalase observed to be induced by Protandim in the previous study [12] do not have ARE sites in their promoter regions. Different components are likely to play a primary role with respect to different end points of oxidative stress. As indicated previously, the composition of Protandim was designed based on the vast amount of studies carried out with those phytochemicals. The in vitro
cell culture model used in this study could be used to determine the role of different components of Protandim on diverse end points. We will be also able to design different combinations of phytochemicals depending on the objective with respect to different disease conditions.
Phosphorylation of Nrf2 on serine 40 results in its dissociation from Keap1 and translocation to nucleus [23]. Inducers of Nrf2-driven phase 2 enzymes have been reported to use multiple signaling pathways for Nrf2 phosphorylation. For example, signaling mediated by PI 3-kinase [13, 24], MEK/ERK [51], p38 MAPK [16], JNK [26] and Protein kinase C [52] have been shown to play a role in the induction of HO-1. In the present study, we observed significant decrease in Protandim-mediated HO-1 induction when PI 3-kinase, Akt, PKCd or p38MAPK was inhibited (Fig. 7). In our previous report with curcumin, we did not observe a significant role for p38 MAPK and PKCd played a minor role in the case of demethoxy curcuminoids [13]. Therefore it
appears that components other than curcumin might be contributing to HO-1 induction through p38 MAPK and PKCd. The concomitant stimulation of parallel signaling pathways seems most likely to be the source of the observed synergy among the components of Protandim.
The findings described in this study suggest that Protandim induces HO-1 through activation of Nrf2 by a mechanism involving multiple signaling pathways. Nrf2 is also known to induce several other antioxidant enzymes including enzymes involved in the synthesis of glutathione. Glutathione synthesis is regulated by g glutamyl cysteine ligase which consists of a regulatory subunit (GCLM) and a catalytic subunit (GCLC). The expression of both GCLM and GCLC is regulated by the Keap1-Nrf2-ARE pathway [53, 54]. Significant increases in the cellular glutathione content were observed in
Protandim-treated cells suggesting the induction of enzyme(s) involved in glutathione synthesis. This observation is of therapeutic significance because glutathione deficiency contributes to oxidative stress and plays an important role in the pathogenesis of many diseases [55].
Several studies have examined a possible link between consumption of diets rich in flavonoids and protection from diseases associated with oxidative stress [56, 57]. However doubts have been raised because of the low plasma concentrations of individual compounds after consumption through diet. These concentrations are significantly low when compared to those used in in vitro studies. Furthermore when higher pharmacological doses are used to demonstrate their effects in vivo, they cause toxic side effects. It is possible that the beneficial effects of dietary phytochemicals could result from the synergy between those compounds when used at low doses. Ayurvedic medicine also suggests synergy between components from one or more
herbal preparations [58]. This possibility is evident from the findings of this study. The dose response of Protandim on HO-1 promoter activity also gives a sigmoidal curve which is a marker for synergy (Fig. 1). Even at 10 mg/ml, Protandim is able to induce the HO-1 promoter. Curcumin, the primary inducer of HO-1, is present at a concentration of 1.05 mg/ml or 2.8 mM in a 10 mg/ml extract of Protandim. This is significantly lower than
the concentration of 20 mM required to demonstrate an noticeable effect is our recent study [13]. The ability of curcumin to induce HO-1 at such low concentrations in the presence other ingredients strongly suggest that there is synergy among the phytochemicals.
http://www.protandim.com/
