Molecular Mechanisms for Curcumin Benefits Against Ischemic Injury

19 07 2010

To the Editor:

Curcumin (diferuloylmethane) is the yellow bioactive pigment from the powdered dry rhizomes of Curcuma longa Linn (commonly known as turmeric). Heretofore, numerous biological targets and pharmacological activities have been reported for curcumin, thereby posing this polyphenol as a promising therapeutic agent against a wide variety of disorders.

A number of studies have reported protective effects of this polyphenol in reducing ischemia reperfusion (I/R) injury in several tissues. Among these studies, is the recent interesting article by Wei et al. (1). In the mentioned study, the authors demonstrated the protective effect of curcumin on testicular I/R injury. Based on their findings, the authors proposed that the protective effects of curcumin could be due to its beneficial impacts on the formation and scavenging of reactive oxygen species (ROS) as well as induction of heme-oxygenase-1. Herewith, I would like to propose 4 other potential mechanisms by which curcumin may exert its protective effects against I/R injury.

There are considerable data indicating the role of pro-inflammatory cytokines such as (TNF)-α, IL-6 and Interleukin-1β in the ischemic injury. These cytokines promote the activation of endothelial cells through upregulation of several surface adhesion molecules which leads to the rolling, adhesion, activation and transmigration of neutrophils in the tissue. Intercellular adhesion molecule-1 (ICAM-1; CD54) is among the most important endothelial cell adhesion molecules which is upregulated during reperfusion injury. ICAM-1 interacts with the integrin lymphocyte function-associated antigen-1 (LFA-1; CD11a/CD18) on activated neutrophils. This interaction is crucial for neutrophil transmigration across endothelium.

1) Several previous studies have implied that curcumin could downregulate the expression of ICAM-1 and therefore might decrease the LFA-1/ICAM-1 interaction (2, 3).

2) This polyphenol has been shown to exert substantial reductions in the cerebral expression levels of pro-inflammatory cytokines such as TNF-α, IL-1β and IL-6, which are involved in the post-ischemic response and neutrophil activation (4, 5).

3) Along with the ICAM-1, the selectin family (including L-, P- and E-selectin) represent another class of cell adhesion molecules which have key roles in mediating neutrophil adhesion, tethering, rolling and activation. E-selectin has been reported to be upregulated on the testicular vascular endothelium following reperfusion (6). Interestingly, there is evidence that curcumin could decrease the expression of E-selectin in cultured human umblical vein endothelial cells (HUVEC) (7). Since curcumin could inhibit the NF-κB signaling pathway, downregulation of ICAM-1 and selectin expression by curcumin might be due to the alterations in the expression of numerous NF-κB-regulated genes including those of various surface adhesion molecules (2, 3).

4) There is also a large body of evidence supporting the involvement and upregulation of matrix metalloproteinases (MMPs), in particular MMP-2 and MMP-9, in the development of I/R injury in different tissues. Accordingly, MMP inhibition has been reported to exert favorable effects against I/R injury. Interestingly, the expression and activity of MMP-2 and MMP-9 has been reported to be decreased by curcumin (8).

These mechanisms, apart from the well-known antioxidant properties of curcumin, should be taken into consideration as mechanisms that may, at least partly, account for the observed protective effects against I/R injury. However, additional studies are required to confirm these mechanisms in the testicular tissue.

Amirhossein Sahebkar, Pharm.D.
Cardiovascular Research Center
Avicenna Research Institute
Biotechnology Research Center and School of Pharmacy
Mashhad University of Medical Sciences (MUMS)
Mashhad, Iran

References
1. Wei SM, Yan ZZ, Zhou J. Curcumin attenuates ischemia-reperfusion injury in rat testis. Fertil Steril 2009;91:271-7.

2. Kumar A, Dhawan S, Hardegen NJ, Aggarwal BB. Curcumin (diferuloylmethane) inhibition of tumor necrosis factor (TNF)- mediated adhesion of monocytes to endothelial cells by suppression of cell surface expression of adhesion molecules and of nuclear factor-κB activation. Biochem Pharmacol 1998;55:775–83.

3. Kunnumakkara AB, Diagaradjane P, Anand P, Harikumar KB, Deorukhkar A, Gelovani J, et al. Curcumin sensitizes human colorectal cancer to capecitabine by modulation of cyclin D1, COX-2, MMP-9, VEGF and CXCR4 expression in an orthotopic mouse model. Int J Cancer 2009;125:2187–97.

4. Gonzales AM, Orlando RA. Curcumin and resveratrol inhibit nuclear factor-kappaB-mediated cytokine expression in adipocytes. Nutr Metab 2008;5.

5. Shehzad A, Lee YS. Curcumin: Multiple molecular targets mediate multiple pharmacological actions – A review. Drugs Fut 2010;35:113-9.

6. Sukhotnik I, Greenblatt R, Voskoboinik K, Lurie M, Coran AG, Mogilner JG. Relationship between time of reperfusion and E-selectin expression, neutrophil recruitment, and germ cell apoptosis after testicular ischemia in a rat model. Fertil Steril 2008;90(4 SUPPL):1517-22.

7. Fuller B, Dijk S, Butler P, Hoang V, Davidson B. Pro-inflammatory agents accumulate during donor liver cold preservation: A study on increased adhesion molecule expression and abrogation by curcumin in cultured endothelial cells. Cryobiology 2003;46:284-8.

8. Kim SY, Jung SH, Kim HS. Curcumin is a potent broad spectrum inhibitor of matrix metalloproteinase gene expression in human astroglioma cells. Biochem Biophys Res Commun 2005;337:510–6.

Published online in Fertility and Sterility doi:10.1016/j.fertnstert.2010.07.1071

The Authors Respond:

Testicular torsion is a common urologic emergency. Surgical detorsion is currently the only treatment for testicular torsion. It appears that the main pathophysiology of testicular torsion-detorsion is ischemia-reperfusion injury of testis. The testicular ischemia-reperfusion injury is associated with overgeneration of reactive oxygen species (ROS) such as superoxide anions, hydrogen peroxide, hydroxyl radicals, nitric oxide, and so on. ROS inflict significant injury on ischemic tissue through oxidation of cell membrane lipids, protein, and DNA.

In our previous work, we have demonstrated the protective effect of curcumin on testicular ischemia-reperfusion injury (1). The protective mechanism of curcumin may be due to inhibition of ROS formation, scavenging of ROS, and induction of heme-oxygenase-1. These mechanisms show the well-known antioxidant properties of curcumin.

We read with great interest the letter by Sahebkar. In the letter, the author proposes 4 other potential mechanisms by which curcumin may exert its protective effects against ischemia-reperfusion injury. In fact, these molecular mechanisms reveal the process of ROS generation and antioxidant properties of curcumin.

One of important sources of ROS generation involves the respiratory burst by neutrophils that infiltrate into tissue itself (2). The endothelial cell adhesion molecules play a critical role in recruiting neutrophils to the endothelium of testicular subtunical venules after testicular ischemia-reperfusion.

One class of endothelial cell adhesion molecules is the selectin (2). The selectin family consists of three members: L, P and E-selectins. E-selectin is expressed exclusively on endothelial cells. Some investigators have found that E-selectin knockout mice exhibit a significant reduction in neutrophil recruitment to endothelium of testicular subtunical venules after ischemia-reperfusion of the testis (2). The result indicates that E-selectin is the key cell adhesion molecule in mediating neutrophil adhesion to endothelium.

Intercellular adhesion molecule-1 (ICAM-1) represents another class of endothelial cell adhesion molecules. ICAM-1 interacts with the integrin lymphocyte function-associated antigen-1 (LFA-1) on neutrophils. The ICAM-1/ LFA-1 interaction is crucial for neutrophil adhesion to endothelial cells. Under normal conditions, the basal expression levels of E-selectin and ICAM-1 are low on endothelial cells. Pro-inflammatory cytokines, in particular, tumor necrosis factor-alpha (TNF-α) and interleukin-1-beta (IL-1β), are increased after testicular ischemia-reperfusion (3). TNF-α and IL-1β upregulate expression of endothelial cell adhesion molecules such as E-selectin and ICAM-1 (2,3).

The endothelial cell adhesion molecules mediate neutrophil adhesion to vascular endothelium (3). Subsequently, the neutrophils secrete matrix metalloproteinase-9 (MMP-9) (4). MMP-9 degrades various components of the extracellular matrix, including collagen and laminin in the basal lamina and zona occludens-1 in the endothelial tight junctions (5). Degradation of these components increases vascular permeability and facilitates neutrophil transmigration across vascular endothelium into tissue (6).

Finally, neutrophils that infiltrate into tissue release ROS that can cause tissue injury (2). Curcumin has been shown to downregulate expression of pro-inflammatory cytokines, endothelial cell adhesion molecules, and MMP-9 (7,8). Therefore, curcumin may reduce neutrophil recruitment to the testis, ROS generation, and testicular injury by downregulation of pro-inflammatory cytokines, endothelial cell adhesion molecules, and MMP-9. However, further studies are required to confirm these mechanisms in testicular ischemia-reperfusion injury model.

Si-Ming Wei, M.D., Ph.D.a,b
Zhi-Zhong Yan, M.Sc.c
Jian Zhou, M.D.d

a Department of Surgery, Zhejiang Medical College
b Department of Urology, Third Affiliated Hospital of Hangzhou City, Zhejiang Chinese Medical University
cDepartment of Reproductive Medicine , Red Cross Hospital of Hangzhou City,
dDepartment of Surgery, First Affiliated Hospital , Zhejiang Chinese Medical University, Hangzhou City, Zhejiang Province, China

References
1. Wei SM, Yan ZZ, Zhou J. Curcumin attenuates ischemia-reperfusion injury in rat testis. Fertil Steril 2009;91:271-7.

2. Lysiak JJ, Turner SD, Nguyen QA, Singbartl K, Ley K, Turner TT. Essential role of neutrophils in germ cell-specific apoptosis following ischemia/reperfusion injury of the mouse testis. Biol Reprod 2001 ;65:718-25.

3. Lysiak JJ, Nguyen QA, Kirby JL, Turner TT. Ischemia-reperfusion of the murine testis stimulates the expression of proinflammatory cytokines and activation of c-jun N-terminal kinase in a pathway to E-selectin expression. Biol Reprod 2003 ;69:202-10.

4. Lindsey M, Wedin K, Brown MD, Keller C, Evans AJ, Smolen J, et al. Matrix-dependent mechanism of neutrophil-mediated release and activation of matrix metalloproteinase 9 in myocardial ischemia/reperfusion. Circulation 2001 ;103:2181-7.

5. Rosenberg GA, Cunningham LA, Wallace J, Alexander S, Estrada EY, Grossetete M, et al. Immunohistochemistry of matrix metalloproteinases in reperfusion injury to rat brain: activation of MMP-9 linked to stromelysin-1 and microglia in cell cultures. Brain Res 2001 ;893:104-12.

6. Gidday JM, Gasche YG, Copin JC, Shah AR, Perez RS, Shapiro SD, et al. Leukocyte-derived matrix metalloproteinase-9 mediates blood-brain barrier breakdown and is proinflammatory after transient focal cerebral ischemia. Am J Physiol Heart Circ Physiol 2005 ;289:H558-68.

7. Aggarwal BB, Sung B. Pharmacological basis for the role of curcumin in chronic diseases: an age-old spice with modern targets. Trends Pharmacol Sci 2009 ;30:85-94.

8. Fuller B, Dijk S, Butler P, Hoang V, Davidson B. Pro-inflammatory agents accumulate during donor liver cold preservation: a study on increased adhesion molecule expression and abrogation by curcumin in cultured endothelial cells. Cryobiology 2003 ;46:284-8.

Published online in Fertility and Sterility doi:10.1016/j.fertnstert.2010.07.1070

Advertisements

Actions

Information

2 responses

14 12 2010
Bing H. Tang, M.D. M.P.H.

I recently read with interest the letter by Sahebkar. In the letter, the author proposes 4 other potential mechanisms by which curcumin may exert its protective effects against ischemia-reperfusion injury. In fact, these molecular mechanisms reveal the process of ROS generation and antioxidant properties of curcumin.

Indeed, Sahebkar’s proposal is not only interesting but also important on molecular base. In fact, other than curcumin, this author would like to propose as following:

OXIDANT-INDUCED VEGF-EXPRESSION OF STEM CELLS AUGMENTED WITH THE EXTRACT OF GRAPE SEEDS PROANTHOCYANIDINS

Vascular endothelial growth factor (VEGF) is the most prevalent, efficacious, and long-term signal that is known to stimulate in both wounds and surgical sites.

Proanthocyanidins are biologically active polyphenolic bioflavonoids.
Grape seed proanthocyanidin containing resveratrol (GSPE) facilitates oxidant-induced VEGF-expression in keratinocytes. GSPE can up-regulate hydrogen peroxide, and advancing stem-cell maturation, which was targeting K562 (CCL-243, ATCC), a human erythroleukemia cell line.

Material and methods:

Among 11 natural health substances, highly augmented CD34CD45+ stems cell suspensions were cultured. The expression of stem-cell mediated cytotoxicity with ratio of survival of K562 cells of GSPE treated groups versus the controls.

Cytotoxicity Assay was done according to the technique of Nociari, MM.

Results:

There were 52 measures having ratio (the sample % vs. the control % of cells gated in the flow cytometry) less than 1.50, while there were merely three greater than 1.50.

The difference between these two groups is statistically significant (two tailed p < 0.005). Flow cytometric analysis of five CDs revealed that among CD 34 hematopoietic stem cells, among those 11 natural health substances, the ratio of GSPE is highest, 2.18.

On the other hand, human umbilical cord blood possesses more progenitor cells than peripheral blood. The UCB samples exhibited a significant immune response on stem cells CD34CD45.

CONCLUSION

After treating the mononuclear cells (MNCs) of human umbilical cord blood (hUCB) for the crude extracts of 11 natural health substances, the extract of grape seeds (GSPE) has the highest actively cellular
differentiation: CD34 can be found on hematopietic stem cells, while
CD45 can be found on all nucleated hematopoietic cells.

The identification of GSPE for augmenting stem cells function may open a new avenue of promoting oxidant-induced VEGF-expression in order to advance new ventures of endovascular therapy, and thereby posing Proanthocyanidins as a promising therapeutic agent against a wide variety of disorders.

A number of studies have reported protective effects of Proanthocyanidins in reducing ischemia reperfusion (I/R) injury in several tissues. Among these studies, is the recent interesting article by Tang et al. (1). In the mentioned study, the authors demonstrated the protective effect of oxidant-induced VEGF-expression.

Bing H. Tang, M.D. M.P.H.
Dr. Chen's Systemic Biology Laboratory
Institute of Biological Chemistry
Academia Sinica
Nankang, Taipei, Taiwan

Research Consultant
New York College of Traditional Chinese Medicine Mineola, New York

Researcher
Research and Ethics
Ranville, California

Reference
(1) Tang, B., Chen S., et al. The effect of grape seed extract on hematopoietic stem cells in the umbilical cord blood. In Vivo, Fall 2007, vol. 29, No. 1 pp 12-15

14 12 2010
Bing H Tang, M D

The crude extract of grape seed was studied, and introduced in our aforeposted response. It was among the following 11 natural health substances:

Quercetin dihydrate, Astaxanthin, Ginsenoside Compund-K (CK-1), Cordyceps Sinensis (TCS-W), Caffeic acid, Bilobalide, Caffeic acid, Eugenol, Rutin hydrate, wheat grass extracts, resveratrol, proanthocyanidin (grape seed extract, GSPE), Ganoderma lucidum.

These 11 substances were as well used in our other study of N K cells as shown in the reference 5] listed below.

Bing H. Tang, M.D. M.P.H.
Dr. S T Chen’s Systemic Biology Laboratory
Institute of Biological Chemistry
Academia Sinica
Nankang, Taipei, Taiwan

Research Consultant
New York College of Traditional Chinese Medicine
Mineola, New York

Researcher
Research and Ethics
Danville, California

References
1] Tang, B., Chen S., et al. The effect of grape seed extract on hematopoietic stem cells in the umbilical cord blood. In Vivo, Fall 2007, vol. 29, No. 1 pp 12-15

2] Tang B, Chen S T et al. Stimulating angiogenesis: Oxidant-Induced
vegf-Expression of stem cells augmented with the extract of grape seeds poanthocyanidins. International Intracranial vascular Surgery Conference, Dec, 3, 2006

3] Tang B. et al. 2007, Vascular endothelium Growth Factor of Stem Cells Enhanced with Proanthocyanidins. International Symposium of Pediatric Neuro-Oncology, October, 21, 2007

4] Tang B. et al, 2007, Vascular endothelial growth factor (VEGF) in
Neurosurgery: in association with the effect of grape seed extract on
hematopoietic stem cells in the umbilical cord blood. November 24

5] Tang B, Chen S T et al. In vitro Maturation Of NK Cells In The Human Umbilical Cord Blood Treated With Wheat Grass Extract: A Pilot Study In Vivo, Vol. 29 (3) 9-20

Leave a Reply

Fill in your details below or click an icon to log in:

WordPress.com Logo

You are commenting using your WordPress.com account. Log Out / Change )

Twitter picture

You are commenting using your Twitter account. Log Out / Change )

Facebook photo

You are commenting using your Facebook account. Log Out / Change )

Google+ photo

You are commenting using your Google+ account. Log Out / Change )

Connecting to %s




%d bloggers like this: