Dietary Milk Phospholipids Increase Body Fat and Modulate Gut Permeability, Systemic Inflammation, and Lipid Metabolism in Mice

The study aimed at how dietary milk polar lipids affect gut permeability, systemic inflammation, and lipid metabolism during diet-induced obesity (DIO). C57BL/6J mice (n = 6x3) were fed diets with 34% fat as energy for 15 weeks: (1) modified AIN-93G diet (CO); (2) CO with milk gangliosides (GG); (3) CO with milk phospholipids (MPL). Gut permeability was assessed by FITC-dextran and sugar absorption tests. Intestinal tight junction proteins were evaluated by Western blot. Plasma cytokines were measured by immunoassay. Body composition was assessed by magnetic resonance imaging. Tissue lipid profiles were obtained by thin layer chromatography. Hepatic expression of genes associated with lipid metabolism was assessed by RT-qPCR. MPL increased the efficiency of converting food into body fat and facilitated body fat accumulation compared with CO. MPL and GG did not affect fasting glucose or HOMA-IR during DIO. MPL increased while GG decreased plasma TG compared with CO. MPL decreased phospholipids subclasses in the muscle while increased those in the liver compared with CO. GG and MPL had little effect on hepatic expression of genes associated with lipid metabolism. Compared with CO, MPL decreased polar lipids content in colon mucosa. Small intestinal permeability decreased while colon permeability increased and then recovered during the feeding period. High-fat feeding increased plasma endotoxin after DIO but did not affect plasma cytokines. MPL and GG did not affect plasma endotoxin, adipo-kines and inflammatory cytokines. After the establishment of obesity, MPL increased gut permeability to large molecules but decreased intestinal absorption of small molecules while GG tended to have the opposite effects. MPL and GG decreased mannitol and sucralose excretions, which peaked at d 45 in the CO group. MPL decreased occludin in jejunum mucosa compared with CO. GG and MPL did not affect zonula occludens-1 in gut mucosa. In conclusion, during DIO, milk GG decreased gut permeability, and had little effect on systemic inflammation and lipid metabolism; MPL facilitated body fat accumulation, decreased gut permeability, did not affect systemic inflammation.


INTRODUCTION
Excess energy can result in body fat accumulation (Saito et al., 2019, Tysoe, 2024) and eventually lead to obesity (Goetzman et al., 2024).Excessive fat accumulation in adipocytes may initiate inflammation since lipid mediators are precursors to inflammatory signaling molecules (Kolb, 2022).The inflamed adipose tissue can produce proinflammatory cytokines, such as tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) (Sethi and Hotamisligil, 2021).These proinflammatory cytokines can initiate local intestinal inflammation and disrupt intestinal barrier (Fang et al., 2023).Obesity, adipose inflammation and compromised gut integrity can self-perpetuate (Wu and Ballantyne, 2020).Another mechanism for leaky gut (Hasegawa et al., 2023, Mishra et al., 2023) in obese animals is the adaptation for nutrient absorption by intestinal hyperplasia (Morton and Hanson, 1984), which causes tight junction (TJ) proteins to malfunction (Ferraris and Vinnakota, 1995).Increased gut barrier permeability can result in endotoxemia and metabolic inflammation (Violi et al., 2023).Mice with diet-induced obesity (DIO) develop endotoxemia compared with lean mice (Naito et al., 2010).High fat diets can independently increase gut permeability and result in endotoxemia that leads to metabolic inflammation (Yan et al., 2024), which may promote obesity (Li et al., 2022).According to current evidence, gut permeability increase, endotoxemia, systemic inflammation and body fat accumulation are all players during the development of DIO.Yet, it is currently unclear which, if any, components drive the process.
Various studies have explored physiological effects of milk polar lipids supplemented in rodent and human diets.Milk sphingolipids reduce the uptake of cholesterol (Eckhardt et al., 2002, Noh andKoo, 2004), protect against bacterial infections in the gut (Vesper et al., 1999, Pfeuffer and Schrezenmeir, 2001, Kosmerl et al., 2023), and reduce the inflammatory response (Park et al., 2005, El Alwani et al., 2006, Park et al., 2007, Dalbeth et al., 2010).Sphingomyelin (SM) affects neonatal gut maturation in rats (Motouri et al., 2003) and regulates intestinal cholesterol absorption (Zhang et al., 2014).Dairy gangliosides (GG) inhibit degradation of gut occludin TJ protein during lipopolysaccharide (LPS) -induced acute inflammation (Park et al., 2010).Milk phospholipids (MPL) reduce hepatic triglycerides (TG) and cholesterol in rats (Cohn et al., 2008).A PL-rich milk fat globule membrane (MFGM) extract reduces hepatomegaly, hepatic steatosis and hyperlipidemia in mice (Wat et al., 2009).A buttermilk MFGM isolate promotes intestinal barrier integrity against LPS stress in mice (Snow et al., 2011).Extracted milk phospholipids reduce de novo hepatic fatty acid biosynthesis (Reis et al., 2013).Dietary milk polar lipids have limited beneficial effects on gut barrier integrity, systemic inflammation, and lipid metabolism in the context of severe obesity (Zhou and Ward, 2019).Milk GG protected the intestinal barrier integrity while MPL had beneficial effect on hepatic lipid metabolism during acute and chronic inflammation induced by lipopolysaccharide (Zhou and Ward, 2024).Based on these findings, it is conceivable that milk polar lipids may influence endotoxemia and systemic inflammation through affecting intestinal barrier integrity.Milk polar lipids could also influence lipid metabolism and the development of DIO.Dietary supplementation of milk polar lipids during the development of DIO may facilitate the understanding of the interrelationships among intestinal barrier integrity, endotoxemia, systemic inflammation and obesity.This study was designed to test the hypotheses that dietary milk polar lipids will reduce liver lipids and affect hepatic expression of genes associated with fatty acid synthesis and cholesterol, prevent gut permeability increase, and reduce systemic inflammation during the development of DIO in C57BL/6J mice fed a diet with 34% fat by energy.

Diets Formulation
Three diets based on the AIN-93G rodent diet were formulated (Zhou and Ward, 2019) so that they were identical in macro and micro nutrient except for the fat source, which was provided by soybean oil + lard (CO diet), soybean oil + lard + milk GG (GG diet), soybean oil + lard + MPL (MPL diet), respectively.The fat provided 34% energy.This amount of fat is considered high compared with the 17.2% (energy) fat in AIN-93G diet.The fatty acid profiles of the diets were reported previously (Zhou and Ward, 2019).The milk polar lipids were provided as a semi-purified MPL concentrate or a semi-purified milk GG concentrate prepared from dried milk cream by ethanol extraction (Fonterra USA Inc., Rosemont, IL).The GG was supplemented at 0.2 g/kg diet, including 0.17 g of ganglioside GD3 and 0.03 g of ganglioside GM3.The GG diet also contained small amount of phosphatidylcholine (PC), phosphatidylserine (PS), phosphatidylethanolamine (PE) and sphingomyelin (SM) (0.55, 0.48, 0.21 and 0.2 g/kg diet, respectively).The MPL was supplemented at 10 g/kg diet, including 2.9 g of SM, 5 g of PC, 1.6 g of PE and 0.6 g of PS (Zhou and Ward, 2024).

Animals
Five-week-old male C57BL/6J mice (n = 18; Jackson Laboratory) were housed in single cages at a constant temperature of 22 ± 1°C with a 12-h light/dark cycle.They were allowed ad libitum access to diet and water.After being fed chow diet for 2 weeks (for acclimatization and baseline data collection), the mice were randomly assigned to one of the following treatments: 1) CO diet (n = 6); 2) GG diet (n = 6); 3) MPL diet (n = 6).The mice were fed the experimental diets for 15 weeks.Diet intake was monitored daily and body weight was measured every other day.Body compositions were assessed every week by magnetic resonance imaging (MRI) using an EchoMRI-900 Body Composition Analyzer (EchoMRI, Houston, TX).The experimental timeline is shown in Figure 1a.The experiments were conducted in conformity with the Public Health Service Policy on Humane Care and Use of Laboratory Animals and were approved by the Utah State University Institutional Animal Care and Use Committee.The protocol number was 1507.

Tissue Sample Collection
The mice were sacrificed by CO 2 asphyxiation after a 4h fast.After blood collection, liver, quadriceps muscle, intestinal and colonic mucosa, feces and adipose tissue samples were collected.The adipose depots included gonadal, retroperitoneal, mesenteric, and subcutaneous depots.Each category of tissue was saved separately and the tissue mass was recorded.

Liver Gene Expression Analysis
The hepatic expression of genes associated with lipid metabolism was analyzed by real-time quantitative polymerase chain reaction (RT-qPCR) assays.Total RNA was extracted and converted into cDNA as previously described (Zhou et al., 2012).RT-qPCR was then performed with the EvaGreen® method using Biomark 48.48 Dynamic Arrays (Fluidigm, South San Francisco, CA).Primer sequences were as previously described (Zhou and Ward, 2019).The cycle threshold (Ct) values for the genes of interest were normalized with the Ct values for peptidylprolyl isomerase A. The relative gene expression was calculated by using the 2 −ΔΔCt method.

GG Analysis of Intestinal Mucosa
The total GG content in intestinal mucosa was determined by measuring the gangliosides bound sialic acid with gas chromatography-mass spectrometry (GC-MS).The gangliosides were extracted and purified by using Sep-Pak C18 reverse-phase cartridges (Waters, Milford, MA) according to method of Schnabl et al. (Schnabl et al., 2009).The gangliosides were derivatized by trimethylsilylation according to method of Carter and Gaver (Carter and Gaver, 1967).The samples were analyzed by a Shimadzu GC-2010 coupled with a Mass Spectrometer.The quantification was achieved from the standard curve generated by concurrent analysis of a series of ganglioside GD3 standards in different concentrations.

Statistical Analyses
The plasma FITC data and HPTLC intensity data were log-transformed to reduce skewness.One-way or mixed models ANOVA (ANOVA) was performed by SAS 9.2.The group means were compared by Ryan-Einot-Gabriel-Welsch Multiple Range Test or Least Squares Means Contrast in SAS.The data were reported as Mean ± Standard Error of the Mean (SEM).

Food Intake and Body Growth
The MPL group consumed more diet than the other 2 groups during the first 3 d and the difference persisted until d 10 (Supplementary Table 1).The MPL group consumed 1.3 g more diet during the first day than the CO group and 0.97 g more diet than the GG group.The daily food intake for the first 3 d was larger than during the whole study in all groups.There were no statistically significant differences among groups regarding diet intake during the rest of the study (Figure 1b).There was no overall treatment effect but significant time effect on body weight during the study (Figure 1c).There was a significant diet effect for body weight gain (P = 0.02) during the first 3-10 d of dietary treatment (Supplementary Figure 1).The MPL group consumed more energy, gained more body fat and converted a higher percentage of consumed energy into body fat from 3 d before to 4 d after dietary intervention (Table 1).The difference was no longer significant by the end of the study for total energy consumption (Table 1).The MPL group accumulated more body fat and had a higher conversion rate from energy to body fat (only statistically significant by posthoc tests).
The body fat composition data (by MRI) indicated that the percent body fat reached a plateau at around d 68 (Figure 1d and e).There was significant time effect on body lean mass during the study (Figure 1d and f).The plots of body fat against fat-free mass showed that the slope increased gradually until the data points segregated at d 68 (Supplementary Figure 2a-f), which indicated the establishment of the DIO (Fenton, 1956).The MPL facilitated fat accumulation after d 68 compared with the CO and GG (Figure 1d and e).The body fat percentage of the MPL group increased at a faster rate compared with that of the other groups (Figure 1e).The accumulating body weight gain of the MPL group tended to increase at a higher rate compared with the other groups but the slope was much smaller (Figure 1c) than that of the body fat increase (Figure 1d).There was no diet effect on liver mass or liver mass normalized to body weight (Supplementary Table 2 online).There were no differences in tissue masses for skeletal muscle and adipose depots (Supplementary Table 2 online).

Plasma Glucose and HOMA-IR
Hyperglycemia (fasting glucose >250 mg/dl (Clee and Attie, 2007)) was observed at d 101 (Figure 2a).The HOMA-IR index increased over time (Figure 2b).By the end of the study, insulin resistance was developed as indicated by HOMA-IR (Lemaitre et al., 2018).Dietary polar lipids did not affect plasma glucose and HOMA-IR.

Plasma and Tissue Lipid Profiles
Plasma TG increased over time and the MPL group had higher plasma TG (Figure 2c) compared with the CO and GG groups at the end of the study.Plasma CE increased over time during the study and there was no treatment effect (Figure 2d).Plasma FFA (Supplementary Figure 3) and PC (Figure 2e) were lower in the MPL and GG groups compared with that in the CO group at d 35.Those differences disappeared toward the end of the study.Plasma SM decreased over time and the MPL group had higher plasma SM (Figure 2F) compared with the CO and GG groups at the end of the study.Lipid classes were analyzed in gonadal adipose tissue, liver and skeletal muscle.Those analytes that differed between diets are shown in Figure 3.The MPL decreased the adipose PC and SM compared with the CO.The MPL increased the liver PE, PC and SM (Figure 3a) compared with the CO.Dietary treatment did not affect lipid profile in the skeletal muscle.Dietary GG did not affect the mucosal gangliosides content.Dietary MPL increased the mucosal PC in the small intestine and decreased mucosal PC, PE and SM in the colon compared with the CO and the GG (Figure 3b).Expression of 13 genes involved with lipid metabolism was analyzed using RT-qPCR and those genes affected by diet treatment are shown in Figure 3c.The MPL suppressed hepatic expression of the fatty acid synthesis gene Acacb (vs the GG) and upregulated cholesterol reverse transport gene Scarb1 (vs the CO & the GG).

Gut Permeability
Gut permeability was assessed by FITC-dextran absorption test at 3 time points and by DST at 6 time points.Permeability to FITC-dextran decreased over time and there was no treatment effect (diet*time effect: P = 0.013, Figure 4a).The DST revealed significant time effect and diet x time interactions on gut permeability.Mannitol excretion rate increased in the CO and GG groups.Mannitol excretion rate decreased and then increased in the MPL group (Figure 4b).The GG and MPL group had lower mannitol excretion rate compared with the CO group at d 45.That difference lasted until d 75 for the MPL group (Figure 4b).By d 75, the GG group had higher mannitol excretion rate compared with the CO group (Figure 4b).Sucralose excretion rate increased significantly in the CO and GG groups (Figure 4c).Sucralose excretion rate decreased and then increased slightly in the MPL group.The GG and MPL group had lower sucralose excretion rate compared with the CO group at d 45.That difference lasted toward the end of the study(Figure 4c).The urinary lactulose/mannitol ratio decreased over time and there was diet x time effect (time effect: P = 0.001, diet x time effect: P = 0.029, Figure 4d).The MPL increased urinary lactulose/mannitol ratio significantly at d 73 and then decreased it toward the end of the study (Figure 4d).There was time effect but no treatment effect for the urinary sucrose/lactulose ratio (time effect: P < 0.0001, Figure 4e).The urinary sucrose/lactulose ratio decreased toward d 45 before rising again (Figure 4e).The urinary lactulose/sucralose ratio decreased significantly during the first 45 d (time effect: P < 0.0001, Figure 4f).There was diet x time effect (P = 0.036) and the MPL increased the ratio significantly at d 73 (Figure 4f).The urinary sucralose/mannitol ratio in the CO group increased slightly during the first 15 d, decreased toward d 30, increased significantly toward d 45, dropped significantly toward d 73 and then increased toward d 87 (time effect: P < 0.0001, Figure 4g).The MPL group had higher urinary sucralose/mannitol ratio at d 45 compared with the other groups (Figure 4g).As indicated by Western blot (Figure 4h), the MPL decreased tight junction protein occludin in jejunum mucosa but did not affect occludin in the mucosa of ileum and colon compared with the CO.The GG increased the tight junction protein ZO-1 in the colon mucosa compared with the CO and the ZO-1 was not affected in the mucosa of jejunum and ileum (Supplementary Figure 4).

Plasma Endotoxin and Cytokines
Plasma endotoxin and cytokines were measured at 3 time points (Figure 5).Plasma LPS did not change significantly during the first 35 d and then increased significantly after 101 d (from 500 to 2200 EU/ml, time effect: P < 0.0001, Figure 5a).The diet x time effect was almost significant (P = 0.051).The MPL group had higher plasma LPS compared with the CO group at d 101 (Figure 5a).The GG and MPL did not significantly affect plasma MCP-1, TNF-α, IL-6, leptin, resistin, PAI-1 and insulin (Supplementary Table 3 online and Figure 5b).Plasma leptin and resistin increased significantly after 101 d (time effect: P < 0.0001, Figure 5c&d).The MPL increased plasma PAI-1 during the study compared with the other 2 groups (diet effect: P = 0.045, Figure 5e).Plasma insulin increased over time (time effect: P = 0.008, Figure 5f).

DISCUSSION
This study was designed to test the hypotheses that the milk polar lipids (1) reduce liver lipid level and affect the expression of genes associated with fatty acid synthesis and cholesterol regulation in the liver, (2) prevent the increase of gut permeability, and (3) reduce plasma inflammatory cytokines in C57BL/6J mice during the development of DIO.The mouse model of DIO (with hyperglycemia and insulin resistance), increased gut permeability and increased systemic inflammation was successfully established.The second hypothesis was supported by the data.Both MPL and GG prevented gut permeability increase, as indicated by the decrease of mannitol and sucralose excretions.The MPL promoted body fat accumulation and increased obesity.Milk polar lipids only slightly affected hepatic expression of 2 out of 13 genes associated with lipid metabolism.The MPL increased colon permeability and decreased occludin in the jejunum mucosa.The MPL increased plasma LPS and did not affect plasma inflammatory cytokines.The GG  decreased ZO-1 in the colon mucosa and did not affect gut permeability and plasma inflammatory cytokines.
During this study, the main endpoints were measured at 3-6 time points.Although the results only supported a few hypotheses, this study revealed important dynamic changes in body fat and gut permeability and interesting interactions between milk polar lipids, body fat and gut permeability during development of DIO.
The MPL facilitated body fat accumulation by increasing efficiency of converting consumed energy into body fat especially during the first few days (Table 1).During the early phase of obesity in humans, the adipose expansion is mainly due to adipocytes hypertrophy.In the later stage of obesity after the body weight exceeds 170 per cent of the ideal, adipocytes hyperplasia starts to play a role and the degree of the hyperplasia is well correlated with the obesity severity (Lee et al., 2024).So, it is very likely that the MPL facilitated adipocytes hypertrophy at the beginning.The relevant mechanism needs further exploration.Body fat percentage increased at a faster rate in the MPL group from d 55 (Figure 1e).Two hypotheses may be proposed to explain the faster rate.One hypothesis is that phospholipids promote the preadipocyte differentiation and result in the adipocytes hyperplasia in the MPL group.Another hypothesis is that the faster increase in body fat was due to adipocytes hyperplasia when the body weight exceeded a threshold.Lecithin promotes the preadipocyte differentiation, upregulates differentiation-specific gene expression, and increases lipid levels in the adipocytes (Zhang et al., 2009).The L-α-lysophosphatidylinositol is positively associated with obesity in humans (Moreno-Navarrete et al., 2012).At the time when the fat accumulation started to occur at a faster rate in the MPL group (d 55, Figure 1e), the mice were not obese yet.So, the first hypothesis may be better supported.The body fat percentage of the CO group reached a plateau at d 65.At around d 85, the percent body fat plateaued in the GG and MPL groups.Mice in the MPL group had an increase of approximately 240% more body fat on average than mice in the CO group (Figure 1e).The further increase of the body fat after d 65 in the GG and MPL groups could have been caused by the phospholipids in the diets.It is possible that the phospholipids induced the preadipocytes differentiation from the beginning of the dietary treatment.It may be further hypothesized that the phospholipids-induced adipocytes hyperplasia did not contribute significantly to the body fat content and the later hypertrophy of the newly differentiated adipocytes were responsible for the further body fat increase after d 65 in the MPL group.
Plasma LPS did not increase during the first 35 d, whereas it increased from d 35 to 101.Plasma LPS increased significantly in all groups once the DIO was established (Figure 5a).The increase of plasma LPS was accompanied by the increase of colon permeability (Figure 4g).The increase of colon permeability and the establishment of DIO resulted in a big increase of plasma LPS.It is still not clear if plasma LPS increase was mainly due to colon permeability increase or was the result of DIO.There were no significant dietary treatment effects on major plasma inflammatory cytokines.
During the maturation of the gut, permeability decreased as indicated by decreased plasma FITC (Figure 4a) and the urinary lactulose/mannitol ratio (Figure 4d).Compared with the CO and the GG, the MPL facilitated the decrease of permeability to FITC when the mice were lean and increased the permeability to FITC when the animals were obese.After the DIO was established, the small intestinal permeability increased significantly in the MPL group compared with that in the CO and GG groups.There was a peaking of the body fat percentage around that time point (Figure 1e).The opposite effect of the MPL before DIO and after DIO was quite interesting.It is possible that MPL may facilitate gut maturity by providing building material but may increase gut permeability when the animal is obese.
Colon permeability increased during the first 15 d, receded toward d 30, and then increased significantly at d 45 (Figure 4g).The initial increase of colon permeability may be caused by the high fat diet, which increases intestinal permeability through the dietary fat and increased luminal bile juice levels (Suzuki and Hara, 2010).The high fat feeding before the DIO increased the colon permeability but did not increase plasma LPS significantly.
The significant increase of the colon permeability at d 45 (Figure 4g) may be due to the increased inflammation as indicated by the increase of plasma IL-6 (Figure 5b) around this time.The colon permeability decreased (Figure 4g) and plasma LPS increased (Figure 5a) after the establishment of DIO.The increase of plasma LPS after the DIO may not be contributed significantly by the decreased colon permeability but may be mediated through the chylomicrons as the postprandial carriers for the LPS (Lebrun et al., 2022).The chylomicron secretion process is different in DIO from when the mice are lean (Uchida et al., 2012).The mechanisms remain unclear for the altered intestinal TG metabolism in mouse models of obesity (Uchida et al., 2012).Taken together, it may be hypothesized that the absorption of gut LPS in DIO is mainly mediated through the transcellular pathway instead of the paracellular route.If the absorbed gut LPS in plasma during DIO is mainly carried by chylomicron remnants, this may help explain the lack of a considerable inflammatory response corresponding to the increased plasma LPS since the lipoproteins may inactivate LPS (Vreugdenhil et al., 2003).Plasma LPS did not increase (Figure 5a) at d 35 but the IL-6 increased manyfold (Figure 5b).Plasma IL-6 then decreased toward the end of the study while the LPS increased dramatically.These results indicate that the initial increase of plasma LPS upon high fat feeding may trigger an inflammatory response, which recedes over time.Once obesity is established, large amount of gut LPS may be absorbed into plasma but may not pose a strong inflammatory stress.
The MPL increased plasma PAI-1 during the first 35 d and plasma PAI-1 flattened in the MPL group when the mice became obese (Figure 5e).PAI-1 is an adipocytokine and the increased PAI-1 in obesity has been associated with the mediation of obesity, insulin resistance and metabolic syndrome (Ma et al., 2004, Alessi andJuhan-Vague, 2006).Lysophosphatidylcholine in oxidized low-density lipoprotein enhances the PAI-1 expression in mouse 3T3-L1 adipocytes (Kuniyasu et al., 2011).The increase of plasma PAI-1 in the MPL group may be caused by the metabolites of dietary PC and may have contributed to the development of the DIO.

CONCLUSIONS
The major effects of the dietary polar lipids on lipid metabolism, systemic inflammation, and gut permeability during the development of the DIO are summarized in Table 2. MPL increased the efficiency of converting food into body fat and promoted body fat accumulation.MPL and GG did not affect fasting glucose or HOMA-IR during DIO.MPL increased while GG decreased plasma TG compared with CO.MPL decreased phospholipids subclasses in the muscle while decreased those in the liver.After the establishment of obesity, MPL increased gut permeability to large molecules but decreased intestinal absorption of small molecules while GG tended to have the opposite effects.MPL and GG did not affect plasma LPS, adipokines and inflammatory cytokines.In conclusion, dietary milk phospholipids increased obesity, prevented gut permeability increase, and did not affect plasma inflammatory cytokines while gangliosides prevented gut permeability increase to a lesser extent.This study revealed important dynamic changes in gut permeability and body fat content during the development of DIO.Dietary milk phospholipids may prevent gut permeability increase but have unfavorable effects on obesity during the development of high fat diet induced obesity.

Notes
ACKNOWLEDGMENTS This research was supported by the Western Dairy Center at Utah State University.In addition, support was provided by the Utah Agricultural Experiment Station, and was approved as paper #9784.
DATA AVAILABILITY The data used for this article are available upon request.The supplementary data file is available: CONFLICT OF INTEREST The authors declare that there is no conflict of interest.
Zhou and Ward: MILK PHOSPHOLIPIDS MODULATE BODY FAT & GUT INTEGRITY
Zhou and Ward: MILK PHOSPHOLIPIDS MODULATE BODY FAT & GUT INTEGRITY Zhou and Ward: MILK PHOSPHOLIPIDS MODULATE BODY FAT & GUT INTEGRITYTable 2 Compared with control, dietary MPL affected metabolism, gut permeability, and inflammation while dietary GG slightly affected metabolism and gut permeability
5 b-f).Plasma IL-6 increased during the first 35 d and then returned toward baseline level (time effect: P = 0.01, Figure
Zhou and Ward: MILK PHOSPHOLIPIDS MODULATE BODY FAT & GUT INTEGRITY