Is Pork Inflammatory? How It Affects Your Body And Health

The purpose of this study was to find out how well boiled pork powder (BPP) and hot water extract powder (HWEP) from 4 cuts of Landrace, Yorkshire, and Duroc (LYD) meat fight inflammation and free radicals. The highest DPPH radical scavenging activities determined were from BPP of Boston butt (13. 65 M TE) and HWEP of loin (19. 40 M TE) and ham (21. 45 M TE). The 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical scavenging activities of BPP from shoulder ham (39. 28 M TE) and ham (39. 43 M TE) were higher than those of other meat cuts, and ham HWEP had the most ABTS radical scavenging activity. A higher oxygen radical absorbance capacity was determined for BPP from ham (198. 35 M TE) and in HWEP from loin (204. 07 M TE), Boston butt (192. 85 M TE), and ham (201. 36 M TE). Carnosine content of BPP and HWEP from loin and were determined to be 106. 68 and 117. 77 mg/g on a dry basis, respectively. The anserine content of BPP (5. 26 mg/g, dry basis) and HWEP (6. The extract from shoulder ham (79 mg/g, dry basis) had the highest value compared to the other meat cuts. The viability of RAW 264. 7 cells was increased with increasing HWEP from loin and ham treatment. This study also found that HWEP from loin and ham significantly decreased the levels of IL-6 and TNF-α. This effect was dose-dependent. These results suggested that boiled pork and hot water extract of pork have antioxidative and cytokine inhibitory effects.

Oxidative damage generally occurs due to free radicals and reactive oxygen species. Synthetic antioxidants (e. g. , propyl gallate, butylated hydroxytoluene, and butylated hydroxylanisole) are often used to fight free radicals in food and living things (Bouayed and Bohn, 2010). However, the use of these radical scavengers is restricted due to potential health risks (Chi et al. , 2014; Zarei et al. , 2014). Lipopolysaccharides (LPS) and other microbial products can make inflammation worse, which is a major defense against pathogens. Macrophages react very strongly to LPS, and when macrophages are activated by LPS, they make and release more inflammatory mediators, such as tumor necrosis factor-α (TNF-α), which is a pro-inflammatory cytokine (Guastadisegni et al. , 2002). But making too many inflammatory cytokines needs to be controlled because it can cause harmful inflammatory responses like rheumatoid arthritis, septic shock, and other long-term inflammatory diseases (Kim et al. , 2011).

In Korea, pork is one of the most popular meats, and this trend has grown over the past few years. Pork consumption per capita in 2005 was 17. 8 kg and rose to 20. 89 kg in 2013 (Korea Meat Trade Association, 2015). Koreans like pork cuts with a lot of fat, like the belly and Boston butt. They don’t like pork cuts with less fat, like loin and ham. However, consumers have recently had a negative preconception about livestock products (Cross et al. , 2007). People think that the high fat content of meats makes them more likely to get obesity, high blood pressure, heart disease, cancer, and other diseases. (Özvural and Vural, 2008). However, meat is a good source of protein with high biological value. Especially, pork is an excellent source of valuable nutrients such as vitamins, minerals (e. g. , vitamin B12, folic acid, and iron), and antioxidative peptides. These histidine-di-peptides, including carnosine (β-alanyl-L-histidine) and anserine (N-β-alanyl-3-methyl-L-histidine), are useful peptides that fight free radicals. They are found in large amounts in heart muscle, the brain, and the nervous system (Quinn et al. , 1992). These antioxidant di-peptides are obtained from the muscle of animals such as pork, beef, chicken, and fish. Some of their well-known jobs are getting rid of toxins from the body and keeping macrophages’ immune systems in check (Guiotto et al. , 2005). The amount of carnosine in raw loin and ham from Korean native black pigs (KNBP) was found to be 1 in our previous study. 35 mg/mL and 1. 27 mg/mL, respectively (Kim et al. , 2013). Even though pork is good for you in many ways, not many studies have looked at how different cuts of pork affect antioxidation and inflammation. Also, pork is often consumed by various cooking methods such as grilling, boiling, and stewing. Many water soluble compounds in pork can be eluted in the hot water when it cooked. So, the point of this study was to find out what happened to proinflammatory cytokines and radical scavenging activity when boiled pork and hot water extracts of four different cuts were used.

We bought four different cuts of pork from a market in Chuncheon: loin, boston butt, shoulder ham, and ham of LYD (castrated, n=4). All the fat under the skin, between the muscles, and that you could see was taken off each piece of raw meat. The meat was then put in five times distilled water, boiled for one hour, and filtered through cheese cloth. The boiled pork and hot water extracts were separated and lyophilized. Based on the concentration, the lyophilized powder was mixed with distilled water and used as a sample for further testing.

Dipeptide (carnosine and anserine) content

Carnosine and anserine content were analyzed in accordance with the methods described by Gil-Agusti et al. (2008). The Polytron PT-2500 E, Kinematica, Switzerland, was used to mix a 3 g sample with 15 mL of distilled water for 10 minutes at 15,000 rpm. Homogenized samples were centrifuged at 3,000 rpm for 10 min. The slurry was filtered through Whatman No. 1 filter paper. We mixed the filtrate with chloroform and shook it for 30 seconds. After that, we centrifuged it at 3,000 rpm for 10 minutes at 4°C. The supernatant was used for dipeptide content analysis. An Agilent Technologies 1260 Infinity HPLC and a Zorbax Eclipse XDB-C18 column (250×4 cm) were used to analyze the samples. 6 mm, 5 μm, Agilent, USA). A flow rate of 1 mL/min, a column temperature of 25°C, and UV absorbance at 210 nm were used to test the samples. The mobile phase was comprised of 0. 01 M sodium dodecyl sulfate soluble in 0. 01 M sodium dihydrogen phosphate at pH 7. 0, and sodium hydroxide was used for validation. Sample volumes injected were 20 μL. All samples and solvents were filtered through 0. 45 μm membrane filters. Standard carnosine and anserine were purchased from Sigma (USA). The experimental values are indicated linearity, using the area ratio method.

We measured how well BPP and HWEP got rid of DPPH radicals according to Blois’s method from 1958, with a few small changes. Briefly, 0. A 2 mM DPPH solution was mixed with a sample of each concentration in distilled water. The mixture was then vortexed and left to sit at room temperature for 30 minutes. After 30 minutes, a UV/VIS spectrophotometer (SpectraMax M2e, Molecular Devices, USA) was used to measure the absorbance at 517 nm. The absorbance was then expressed as millimolar Trolox equivalents (TE) per gram of low molecular weight sample.

ABTS+ scavenging activity was carried out according to the procedure described by Re et al. (1999). ABTS+ was produced by reacting 14 mM ABTS treated with 4. 9 mM potassium persulfate. Before it was used, the mixture was stirred and left to sit at room temperature in the dark for 12 to 16 hours to make a dark blue solution. The ABTS+ solution (SpectraMax M2e, Molecular Devices, USA) was diluted in 5. 5 mM PBS (pH 7. 4) and equilibrated to 30℃ until the absorbance reached 0. 70±0. 02 at 734 nm. The reactions consisted of 50 μL sample and 950 μL of the ABTS+ radical solution. We used a SpectraMax M2e (Molecular Devices, USA) to measure the absorbance at 30°C after 30 minutes. The millimolar TE per gram of low molecular weight sample was then calculated.

A modified version of the Benzie and Strain (1996) method was used to test the ability to reduce ferric ions. The FRAP reagent was produced and added to acetate buffer (300 mM/L, pH 3. Six), 2,4,6-triazine (TPTZ, 10 mM in 40 mM/HCl) solution, and FeCl3·6H2O (20 mM) were mixed in a way that was 10:1:1 (v/v). The mixture was incubated at 37℃. 25 L of sample and 175 L of FRAP reagent were put into test tubes, and they were heated to 37°C for 5 minutes. The absorbance of FRAP was recorded at 595 nm (SpectraMax M2e, Molecular Devices, USA). The FRAP test for each sample was given in units of mol TE per gram of weight, and the linear calibration curve was used to figure it out.

The oxygen radical absorbance activity assay was performed using a modified version of Gillespie et al. (2007). Fluorescein (80 nM) and 25 L of sample were put in containers and shaken. At the end, each well had 200 μL of solution. The mixture was placed at 37℃ for 15 min. Then, 25 L of AAPH (150 mM) was added to each well. Our lab used a fluorometric spectrophotometer (SpectraMax M2e, Molecular Devices, USA) to measure absorbances at 480 and 520 nm (every 60 minutes) after shaking each set automatically at 37℃. Trolox standards and a blank sample were also used. The ORAC values were found by using the area under the curve (AUC) and regression equations to find the relationship between the TE and the net AUC. The final ORAC values are expressed as mol of TE.

A mouse macrophage cell line, RAW 264. 7, was obtained from American Type Culture Collection (ATCC TIB-71TM, USA). RAW 264. Seven percent of the cells were put in 100 mm tissue culture dishes that had Dulbecco’s modified Eagle’s medium (DMEM) mixed with 10% fetal bovine serum (Welgene, USA) and 1% penicillin/streptomycin (Gibco, USA). RAW 264. 7 cells were maintained at 37℃ in a humidied incubator containing 5% CO2.

RAW 264. 7 cells were plated at a density 5×104 cells/mL in tissue culture plates and incubated for 24 h. Five microliters of the sample at 10, 50, 100, 200, and 500 μg/mL concentrations were added to the wells. After an hour, the cells were stimulated with LPS (1 μg/mL) and incubated for 24 h at 37℃. One hundred microliters of MTT (5 mg/mL) was added to each well, and the plates were left to sit at 37°℃ for 4 hours. The medium with MTT was removed and then dissolved in dimethyl sulfoxide (DMSO). The optical density (OD) of each well was measured at 540 nm (SpectraMax M2e, Molecular Devices, USA).

Cells were plated density 2×105 cells/mL and grown to confluence in 24-well tissue culture dishes. Cells were cultured in an incubator (37℃ and 5% CO2) for 24 h. The next day, a new serum-free medium was added to the wells, and 5 L of samples were added at different concentrations (10, 50, 100, 200, and 500 μg/mL). After about an hour, LPS (1 μg/mL) was added to the medium to make it work. It was then kept at 37°C for 24 hours. The medium with LPS was collected and centrifuged at 12,000 rpm for 5 min at 4℃. The levels of IL-6 and TNF-α were analyzed by ELISA as per the manufacturer’s instructions. We found IL-6 and TNF-α using the mouse IL-6 BD OptEIATM, ELISA sets (BD PharMingen, USA) and the mouse TNF (Mono/Mono) BD OptEIATM, ELISA sets (BD PharMingen, USA). The coating buffer with capture antibody was put in a 96-well plate and left to sit at 4°C overnight, which is about 16 hours. The capture antibodies were aspirated and washed 3 times. Blocking solution (Assay Diluent) was added and incubated for 1 h at room temperature. Standards (15. 6, 31. 3, 62. Samples (5, 125, 250, 500, and 1,000 pg/mL) were put into the wells and left to sit at room temperature for two hours. The standards and samples were removed and washed 6 times. Working Detector (Detection Antibody SAv-HRP reagent) was put into each well, and the plates were left to sit for one hour. They were then washed six times. To each well, 100 L of substrate solution (tetramethylbenzidine and hydrogen peroxide) was added. The plate was incubated for 30 min at room temperature in the dark. The stop solution (2 N H2SO4) was added to each well and absorbance from 450-540 nm was read.

One-way analysis of variance (ANOVA) was used to look at all the results, following the general linear model steps for SAS software (ver. 9. SAS Institute Inc. , USA). Results are shown as mean values and standard error of the sample. Duncans multiple range tests were used to determine statistical significance between the treatments (p<0. 05).

Carnosine and anserine content of BPP and HWEP of LYD

shows the carnosine and anserine content of BPP and HWEP from the four pork cuts. In terms of carnosine, both BPP and HWEP of loin had much higher amounts of carnosine than the other pork cuts. Particularly, carnosine in HWEP of the four cuts was higher than BPP. The amount of anserine in BPP and HWEP was higher in shoulder ham than in the other pork cuts. It was found by Tinberge and Slump in 1976 that pork had 104–338 mg/100 g of carnosine and 7–16 mg/100 g of anserine. According to Aristoy and Toldrá (1998), the amount of carnosine in four pig muscles (masseter, trapezius, semimembranosus, and longissimus dorsi) was 21 06, 180. 98, 320. 84, and 313. 02 mg/100 g muscle, respectively. Also, anserine contents of those muscles were 6. 09, 10. 68, 17. 56, and 14. 56 mg/100g muscle. They suggested that the caronsine and anserine levels are a good indication of muscle glycolytic activity. Lumber longissimus dorsi (loin) and semimembranosus (ham) were classified as glycolytic muscle due to higher carnosine content. In cooked beef, the carnosine content was 134 mg/100 g of tissue (Park et al. , 2005). There may not be much carnosine in beef because it has a lot of fat. This is because carnosine is mostly found in the cytosol of skeletal muscle, and more fat would mean less skeletal muscle and less carnosine (Park et al. , 2005). They found that the amount of carnosine and anserine in beef leg muscle was 150 mg/100 g of tissue for carnosine and 50 mg/100 g of tissue for anserine. Plowman and Close (1988) also said that the amounts of carnosine and anserine in beef shoulder and rib steak were 341 mg/100 g of tissue and 58 mg/100 g of tissue, respectively. The findings showed that animal muscles have different amounts of carnosine depending on whether they are glycolytic or oxidative.

Any guess on what most anti-inflammatory food is?

FAQ

Does pork make inflammation worse?

What meats are anti-inflammatory?

Are pork chops anti-inflammatory?

Is pork bad for You?

Unfortunately, that popularity comes at a cost. Along with being the most commonly consumed meat in the world, pork may also be one of the most dangerous, carrying some important and under-discussed risks that any consumer should be aware of (1). 1. Hepatitis E

Why is pork not recommended to eat?

This is not true, some cuts like sirloin and pork rump steak, for example, are very healthy, even healthier than beef and chicken. Only the fattest cuts like bacon and crackling should be avoided.

Does meat cause inflammation?

Chronic inflammation is linked to heart disease , diabetes , and arthritis. While lean meats can be a good source of iron and other minerals, red meat and processed meats are more likely to cause inflammation. They’re typically stocked with saturated fat, prompting immune cells to release inflammatory proteins into the bloodstream.

Why do people eat pork?

Pork is why we love BBQ, taco al pastor and schnitzel. As pigs in a blanket or bacon-wrapped dates, it’s one food that makes a party delicious. This well-known red meat from domestic pigs is the most commonly eaten meat in the world, according to the United Nations Food and Agriculture Organization.