Great Plains Announces New Test for Phospholipase A2 (PLA2)

William Shaw, Ph.D., Lab Director and Matt Pratt-Hyatt, Ph.D., Associate Lab Director

The Great Plains Laboratory is excited to announce a new test for PLA2 activity, and we are the only commercial lab currently offering this particular test in urine. PLA2 is elevated in a wide range of inflammatory disorders from multiple sclerosis to cancer. This test can be easily added to the organic acids test to provide powerful new clinical insights and treatments for a variety of serious illnesses.

PLA2 Overview:

Chronic diseases are caused by many different biological imbalances, but they almost all create and have inflammation as a cornerstone. Inflammation plays a major part in most of the disorders that we spend billions of dollars to combat, searching for relief from the pain, swelling, and other symptoms that inflammation causes. Inflammation is the immune system's natural response to infection and injury. Phospholipase A2 (PLA2) is one of the key biochemical factors produced in the inflammation response. It is commonly found in human tissues, as well as insect and snake venom. In normal amounts, PLA2 is involved in remodeling cell membranes and changing cell architecture. In infections, PLA2 can break down the phospholipids in the membranes of bacteria, fungi, and parasites leading to their death. However, inflammation, like many other biological processes often has negative effects. The same phospholipase that attacks infectious agents may also attack the cell membranes of the human host, damaging or killing those cells. In addition, the products of the PLA2 reaction, lysolecithins and free fatty acids (Figure 1) are powerful detergents that have the ability to denature proteins and destroy their biological functions. The lysolecithins produced by PLA2 initiate the pain response.

The most common free fatty acid produced by PLA2 is arachidonic acid which can increase the production of powerful mediators of inflammation called prostaglandins, leukotrienes, and thromboxanes, collectively known as eicosanoids. These mediators play an important role in the generation and maintenance of inflammation in neural cells. In addition, arachidonic acid can be converted to 4-hydroxynonenal (4-NE), which can be very toxic due to covalent modification of important biomolecules including proteins, DNA, and phospholipids containing amino groups. In addition to PLA2 causing local damage, it may be transported by the blood vessels to other parts of the body, causing widespread tissue damage.


Diseases Associated with PLA2

Increased levels of PLA2 have been observed in most systemic inflammatory diseases. Studies have linked elevated PLA2 activity with multiple sclerosis, rheumatoid arthritis, Crohn's disease, pancreatitis, ulcerative colitis, allergies, atherosclerosis and cardiovascular disease, lung, prostate, small intestine, and large intestine cancers, with increased susceptibility to metastases, Candida infection, asthma, autism, chronic pulmonary obstructive disorder (COPD), and sepsis.

What Causes Elevated PLA2?

Phospholipase A2 is produced by the pancreas and released into the small intestine following a fatty meal. Infection or trauma of the pancreas may result in the release of phospholipase into the circulation, causing widespread damage or even death. Activation by viruses of proenzymes of PLA2 within the pancreas instead of, as normally, in the intestine, may cause pancreatitis. Phosholipase may be produced by cells of the immune system in response to bacterial antigens, especially those containing certain lipopolysaccharides (LPS). Allergies, especially those to house dust and cats, have been implicated as a trigger for PLA2 synthesis and release. Venoms from snakes, spiders, and bees contain high amounts of PLA2, which is responsible for much of the toxicity of these venoms. In addition, microorganisms such as Candida albicans and certain Clostridia species produce PLA2 which increases the ability of the microorganism to infect the host. Trauma may also cause significant increases in PLA2 and result in brain injury.

PLA2 and Inflammatory Disease

Research has implicated PLA2 in the pathophysiology of neurodegenerative diseases such as multiple sclerosis (MS) and Alzheimer's disease (AD). Multiple sclerosis involves both antigen-specific mechanisms and components of the innate immune system that result in inflammatory response. Elevated PLA2 activity was found to be ongoing among MS patients, with the highest levels measured in patients with progressive disease. In the development of Alzheimer's disease, the abnormal PLA2 levels appear to be related to oxidative signaling pathways involving NADPH oxidase and production of ROS species that lead to impairment and destruction of neurons and inflammation of glial cells.

Inflammation is the hallmark of rheumatoid arthritis (RA), a joint-destructive autoimmune disease. PLA2 is found in synovial fluid of RA-affected individuals and in the cartilage of RA patients as compared to cartilage from osteoarthritic and normal individuals.

Measurement of PLA2 is emerging as an important tool for evaluating the chance of cardiovascular disease (CVD), including future stroke, myocardial infarction, heart failure, and other vascular events. PLA2 appears to be more specific than hsCRP for CVD risk and may also have a pivotal role as a mediator of cardiovascular pathology. In atherosclerosis, PLA2 not only activates macrophages and formation of foam cells, but it also hydrolyzes LDL and HDL, spawning increased numbers of pro-atherogenic small LDL particles, and impairing anti-atherogenic HDL. PLA2 activity may even precipitate bleeding from atherosclerotic plaques.

PLA2 is expressed normally in pancreatic, gall bladder, and GI epithelial cells, but is significantly increased in inflammatory gastrointestinal disorders. In ulcerative colitis and Crohn's disease, all intestinal cell types increase expression of PLA2, which increases gut permeability and may actually contribute to infectivity.

PLA2 and Cancer

Elevations of PLA2 have been found in gastrointestinal cancers including colonic adenomas and carcinomas and pancreatic ductogenic carcinomas, among others. Patients with lung tumors positive for PLA2 had a greatly increased tumor growth rate and a markedly reduced survival rate. Patients with lung cancer also had higher plasma levels of PLA2 than patients with benign nodules. A similar pattern has been observed in prostate cancer, although metastatic tumors expressed lower PLA2 than primary tumors. As PLA2 releases arachidonic acid and other fatty acids from cell membranes, they initiate downstream production of tumor-promoting eicosanoids. In cancer, the spread of tumor cells from a primary tumor to the secondary sites within the body is a complicated process involving cell proliferation and migration, degradation of basement membranes, invasion, adhesion, and angiogenesis. Continued research on PLA2 expression in cancer will certainly reveal valuable new insights.

What lowers PLA2?

There has been a great deal of research done by both academia and pharmaceutical companies to find chemical inhibitors to PLA2. However, there has also been research on more natural methods for inhibiting PLA2. Glucocorticoids such as the natural hormone cortisol and pharmaceutical agents such as dexamethasone inhibit the production of phospholipase, decreasing harm caused by the enzyme but also decreasing the benefits of the enzyme in killing harmful microorganisms. Thus, excess glucocorticoids can reduce inflammation in a patient with tuberculosis while reducing the effects of PLA2 against the bacteria resulting in spread of the illness. Lithium at pharmacological doses, carbamazepine, and the antimalarial drug chloroquine are all PLA2inhibitors. Vitamin E is also an inhibitor of PLA2. In addition, eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) (belonging to the omega-3 class of fatty acids) inhibit PLA2. Analysis has shown that treatment with supplements of Cytidine 5'-Diphosphocholine (CDP-choline) can limit the ability of PLA2 to promote inflammation. CDP-choline is a precursor in the formation of phospholipids and has been used as a nutritional supplement at doses ranging from 500-4000 mg per day in the treatment of patients with a variety of disorders including Parkinson's disease, memory disorders, vascular cognitive impairment, vascular dementia, senile dementia, schizophrenia, Alzheimer's disease (especially effective in those with the epsilon-4 apolipoprotein E genotype), head trauma, and ischemic stroke. A trial in patients with Alzheimer's disease indicated that CDP-choline (1,000 mg/day) is well tolerated and improves cognitive performance, cerebral blood perfusion, and the brain bioelectrical activity pattern. No side effects were noticed at the lower doses of CDP-choline and only some mild gastrointestinal symptoms were found using higher doses. No abnormal blood chemistry or hematology values were found after the use of CDP-choline.

Testing for PLA2

Because PLA2 is a relatively small enzyme (about 14 KD), it is able to be excreted in urine. 10 mL of the first morning urine before food or drink is suggested for testing. There are no dietary restrictions. This test is convenient to include with other urine tests such as organic acids, amino acids, and peptides. Since chelating agents might interfere with the test, they should not be used for at least 48 hours prior to testing. PLA2 testing is recommended for the following disorders:

  • Multiple sclerosis
  • Rheumatoid arthritis
  • Crohn's disease
  • Pancreatitis
  • Ulcerative colitis
  • Allergies
  • Cardiovascular disease including atherosclerosis
  • Neurodegenerative diseases
  • Schizophrenia
  • Bipolar depression, subtype with psychosis
  • Candida infection
  • Sepsis
  • Long term depression
  • Asthma
  • Chronic obstructive pulmonary disease (COPD)

Inflammation plays such a key role in so many diseases, and we believe this new PLA2 test will be a valuable tool in the treatment of patients suffering from numerous disorders. The test is now available and we hope you will integrate it into your practice. For more information about PLA2 and possible treatments, please see the references below.

Clinical References:

  • Green, .J.A., Smith, G.M., Buchta, R., et al. (1991) Circulating phospholipase A2 activity associated with sepsis and septic shock is indistinguishable from that associated with rheumatoid arthritis. Inflammation 15: 355-367
  • Lilja, I., Smedh, K., Olaison, G., et al. (1995) Phospholipase A2 gene expression and activity in histologically normal ileal mucosa and in Crohn's ileitis. Gut. 37: 380-385.
  • Mounier, C.M., Wendum, D., Greenspan, E., et al. (2008) Distinct expression pattern of the full set of secreted phospholipases A2 in human colorectal adenocarcinomas: sPLA2-III as a biomarker candidate. Br J Cancer 98: 587-595.
  • Nicolas, J.P., Lin, Y., Lambeau, G., et al. (1997). Localization of structural elements of bee venom phospholipase A2 involved in N-type receptor binding and neurotoxicity. J Biol Chem. 272: 7173-7181.
  • Pinto, F., Brenner, T., Dan, P., et al. (2003) Extracellular phospholipase A2 inhibitors suppress central nervous system inflammation. Glia 44(3):275-282.
  • Pruzanski. W., Scott, K., Smith, G., et al. (1992) Enzymatic activity and immunoreactivity of extracellular phospholipase A2 in inflammatory synovial fluids. Inflammation 16: 451-457.
  • Sawada, H., Murakami, M., Enomoto, A., et al. (1999) Regulation of type V phospholipase A2 expression and function by proinflammatory stimuli. Eur J Biochem 263:826–835.
  • Adibhatla, R.M. and Hatccher, J.F. (2005). Cytidine 5'-Diphosphocholine (CDP-Choline) in Stroke and Other CNS Disorders. Neurochemical Research. 30: 15-23.
  • Shaw, W. Possible synergistic effects of nonesterified fatty acids and lysolecithins, a toxic methionine metabolite, and ammonia in the production of hepatic encephalopathy and schizophrenia. Orthomolecular Medicine. 1988.3: 87.
  • Shaw, W. Possible role of lysolecithins and nonesterified fatty acids in the pathogenesis of Reye's syndrome, sudden infant death syndrome, acute pancreatitis, and diabetic ketoacidosis. Clin. Chem. 1985. 31:1109.
  • Alvarez, X.A., Mouzo, R., Pichel, V., Pérez P., et al. Double-blind placebo-controlled study with citicoline in APOE genotyped Alzheimer's disease patients. Effects on cognitive performance, brain bioelectrical activity and cerebral perfusion. Methods Find Exp Clin Pharmacol. 1999 Nov;21(9):633-44.
  • Fioravanti, M. and Yanagi, M. Cytidinediphosphocholine (CDP-choline) for cognitive and behavioural disturbances associated with chronic cerebral disorders in the elderly. Cochrane Database Syst Rev. 2005 Apr 18;(2):CD000269.
  • Rosenson, R.S. and Gelb, M.H. Secretory phospholipase A2: A multifaceted family of proatherogenic enzymes. Current Cardiology Reports 2009, 11:445–451
  • Farooqui, A.A., Ong, W., and Horrocks, L.A. Inhibitors of brain phospholipase A2 activity: Their neuropharmacological effects and therapeutic importance for the treatment of neurologic disorders. Pharmacol Rev 58:591–620, 2006
  • Wang, M., Hao, F.Y., J.G. Wang, and Xiao, W. Group IIa secretory phospholipase A2 (sPLA2IIa)and progression in patients with lung cancer. European Review for Medical and Pharmacological Sciences 2014; 18: 2648-2654
  • Pniewska, E., Sokolowska, M., Kupry-Lipinska, I., et al. The step further to understand the role of cytosolic phospholipase A2 alpha and group X secretory phospholipase A2 in allergic Inflammation: pilot study. Biomed Red Int. Volume 2014, Article ID 670814, 9 pages
  • Touqui, L. and Alaoui-El-Azher, M. Mammalian secreted phospholipases A2 and their pathophysiological significance in inflammatory diseases. Current Molecular Medicine 2001, 1, 739-754 739
  • Putignano, S., Gareri, P., Castagna, A., et al. Retrospective and observational study to assess the efficacy of citicoline(CDP-choline) in elderly patients suffering from stupor related to complex geriatric syndrome. Clinical Interventions in Aging 2012:7 113–118.
  • Farooqui, A.A. and Horrocks, L.A. Phospholipase A2-generated lipid mediators in the brain: The good, the bad, and the ugly. The Neuroscientist, Vol. 12, No. 3, 245-260 (2006)
  • Mahmoudabadi, A.Z., Zarrin, M., and Miry, S. Phospholipase activity of Candida albicans isolated from vagina and urine samples. Jundishapur J Microbiol. 2010; 3(4): 169-73.
  • Bell , J.G., MacKinlay , E.E., Dick, J.R., et al. Essential fatty acids and phospholipase A-2 in autistic spectrum disorders. Prostaglandins, Leukotrienes and Essential Fatty Acids Volume 71, Issue 4 , October 2004, Pages 201-204
  • Tavares, H., Yacubian, J., Talib, L.L., et al. Increased phospholipase A2 activity in schizophrenia with absent response to niacin. Schizophr Res. 2003 May 1;61(1):1-6.
  • Ross, B.M. , Hughes, B. , Kish, S.J. , and Warsh, J.J. Serum calcium-independent phospholipase A2 activity in bipolar affective disorder. Bipolar Disord. 2006 Jun;8(3):265-70.
  • Eckert, G.P. , Schaeffer, E.L., Schmitt, A., et al. Increased brain membrane fluidity in schizophrenia. Pharmacopsychiatry 2011 Jun;44(4):161-2.
  • Titsworth, W.L., Liu, N.K., and Xu, X.M. Role of Secretory phospholipase A2 in CNS inflammation: Implications in traumatic spinal cord injury. CNS Neurol Disord Drug Targets 2008 June ; 7(3): 254–269
  • Funakoshi, A., Yamada, Y., Migita, Y., and Wakasugi, H. (1993). Simultaneous determinations of pancreatic phospholipase A2 and prophospholipase A2 in various pancreatic diseases. Dig Dis Sci 38: 502-506

The Role of Vitamins, Antioxidants, and Anti-Inflammatories in Breast Cancer Prevention and Treatment

Terri Hirning

October is Breast Cancer Awareness Month. As such, we would like to take a moment to focus on how nutritional and supplement therapy can play a role in the prevention and treatment of cancer. When we look at how antioxidants impact cancer, we can see that there is scientific documentation of reduced development of breast cancer in those with high dietary intake of antioxidants. One study in late 2014 titled The Rotterdam Study provides this information: "These results suggest that high overall dietary antioxidant capacity are associated with a lower risk of breast cancer."1 Women who had higher rates of antioxidant intake via diet were less likely to develop breast cancer. What about those who already had breast cancer? Could it help with treatment? A March 2014 issue of Anticancer Research featured a study showing the use of lycopene and beta-carotene in cell death of human breast cancer cell lines. "Our findings show the capacity of lycopene and beta-carotene to inhibit cell proliferation, arrest the cell cycle in different phases, and increase apoptosis."2 Vitamin C has also been studied in terms of its potential impact on breast cancer deaths and has been shown to have a positive effect on mortality rates. "Dietary vitamin C intake was also statistically significantly associated with a reduced risk of total mortality and breast cancer-specific mortality."3

If we can look at the data and determine that higher intake of antioxidants and nutrients can not only reduce development of breast cancer but can also positively impact the mortality rates of cancer, the question then becomes how do we encourage our patients and clients to incorporate these into their diets with higher frequency? We must educate them on the role antioxidants play and the resources available to obtain them, whether from foods or supplements. For example, lycopene is a nutrient that is highlighted for its anticancer properties, specifically in reference to breast cancer. Lycopene is a carotenoid that gives many fruits and vegetables their red color. Unlike other carotenes, lycopene does not get converted into vitamin A. The top 10 sources of dietary lycopene are:

  • Guava
  • Watermelon
  • Tomatoes (cooked)
  • Papaya
  • Grapefruit
  • Sweet Red Peppers (cooked)
  • Asparagus (cooked)
  • Red (purple) cabbage
  • Mango
  • Carrots

Continued from BioMed Today:

Encouraging patients to incorporate more foods with lycopene, like those listed above, into their diets is one component. Supplements can also be suggested as a potential option. This is especially true in the case of vitamin D, for which there are many studies showing its role in the prevention of cancers. Adequate vitamin D is being revealed as a critical factor for preventing many diseases, including breast cancer, today.8 "Case-control studies and laboratory tests have consistently demonstrated that vitamin D plays an important role in the prevention of breast cancer."9 Unfortunately due to a variety of reasons, many people are deficient in vitamin D which then can then compromise optimal health. Testing for vitamin D levels, ideally twice a year, is a great way to monitor this critical nutrient and help your patients optimize their health and wellness. Supplementation can then also be recommended to optimize levels. Another promising resource for warding off disease and cancer is curcumin, the extract of the turmeric root. Because of its potent antioxidant and antimicrobial properties, it is being studied extensively for its potential in cancer treatment. The American Cancer Society's website has this to say about it: "Curcumin can kill cancer cells in laboratory dishes and also slows the growth of the surviving cells. Curcumin has been found to reduce development of several forms of cancer in lab animals and to shrink animal tumors."4 The typical therapeutic dose, between 3 and 10 grams per day, exceeds what is normally used in cooking and obtained through dietary consumption so a supplement would be most effective.

Could another reason for the efficacy of curcumin on cancer cell death be its potent anti-inflammatory properties? Curcumin has been studied widely for both its safety and anti-inflammatory potential.5,6 "The laboratory studies have identified a number of different molecules involved in inflammation that are inhibited by curcumin including phospholipase, lipooxygenase, cyclooxygenase 2, leukotrienes, thromboxane, prostaglandins, nitric oxide, collagenase, elastase, hyaluronidase, monocyte chemoattractant protein-1 (MCP-1), interferon-inducible protein, tumor necrosis factor (TNF), and interleukin-12 (IL-12)."7 We see science validating the role our lifestyle has in development of cancer. Diet, exercise, supplementation, our stress level, and other factors all contribute to the whether or not we develop disease and also to our ability to reverse it. It is important to find ways to offer a variety of prevention and treatment options that work with our patients' lifestyles.

Clinical References:

  • Pantavos A, Ruiter R, Feskens E, E deKeyser C, Hofman A, H Stricker B, H Franco O, C Kiefte-deJong J (2014). Total dietary antioxidant capacity, individual antioxidant intake and breast cancer risk: The rotterdam study, International Journal of Cancer. 2014 Oct 4. doi: 10.1002/ijc.29249. [Epub ahead of print]
  • Gloria NF, Soares N, Brand C, Oliveira FL, Borojevic R, Teodoro AJ (2014).Lycopene and beta-carotene induce cell-cycle arrest and apoptosis in human breast cancer cell lines, Anticancer Research. 2014 Mar;34(3):1377-86.
  • Harris HR, Orsini N, Wolk A (2014). Vitamin C and survival among women with breast cancer: a meta-analysis,European Journal ofCancer. 2014 May;50(7):1223-31. doi: 10.1016/j.ejca.2014.02.013. Epub 2014 Mar 7.
  • Turmeric (2012). Retrieved on October 5, 2014 from Link
  • Chainani-Wu NJ (2003). Safety and anti-inflammatory activity of curcumin: a component of tumeric (Curcuma longa), Journal of Alternative and Complementary Medicine. 2003 Feb; 9(1):161-8.
  • Jurenka, JS (2009). Anti-inflammatory Properties of Curcumin, a Major Constituent of Curcuma longa: A Review of Preclinical and Clinical Research, Alternative Medicine Review. Volume 14, Number 2, 2009.
  • Nita Chainani-Wu (2003). The Journal of Alternative and Complementary Medicine. February 2003, 9(1): 161-168. doi:10.1089/107555303321223035.
  • Vitamin D and Cancer Prevention (2013). Retrieved on October 5, 2014 from
  • Walentowicz-Sadłecka M, Sadłecki P, Walentowicz P, Grabiec M (2013). The role of vitamin D in the carcinogenesis of breast and ovarian cancer, Ginekologia Polska. 2013 Apr;84(4):305-8.