Potential conflict of interest: Nothing to report.
We recently showed that the heparan sulfate proteoglycan syndecan-1 mediates hepatic clearance of triglyceride-rich lipoproteins in mice based on systemic deletion of syndecan-1 and hepatocyte-specific inactivation of sulfotransferases involved in heparan sulfate biosynthesis. Here, we show that syndecan-1 expressed on primary human hepatocytes and Hep3B human hepatoma cells can mediate binding and uptake of very low density lipoprotein (VLDL). Syndecan-1 also undergoes spontaneous shedding from primary human and murine hepatocytes and Hep3B cells. In human cells, phorbol myristic acid induces syndecan-1 shedding, resulting in accumulation of syndecan-1 ectodomains in the medium. Shedding occurs through a protein kinase C–dependent activation of ADAM17 (a disintegrin and metalloproteinase 17). Phorbol myristic acid stimulation significantly decreases DiD (1,1′-dioctadecyl-3,3,3′,3′-tetramethylindodicarbocyanine perchlorate)-VLDL binding to cells, and shed syndecan-1 ectodomains bind to VLDL. Although mouse hepatocytes appear resistant to induced shedding in vitro, injection of lipopolysaccharide into mice results in loss of hepatic syndecan-1, accumulation of ectodomains in the plasma, impaired VLDL catabolism, and hypertriglyceridemia. Conclusion: These findings suggest that syndecan-1 mediates hepatic VLDL turnover in humans as well as in mice and that shedding might contribute to hypertriglyceridemia in patients with sepsis. (HEPATOLOGY 2012)
Hypertriglyceridemia is a common disorder that results from the accumulation of triglyceride-rich lipoproteins (TRLs) in the circulation. TRLs include chylomicrons derived from dietary lipids, very low density lipoproteins (VLDLs) from the liver, and the remnant particles that remain after the action of lipoprotein lipase in the vasculature. These remnant lipoproteins are cleared from the circulation via endocytic receptors in the liver, including the low density lipoprotein receptor (LDLR),1, 2 the LDLR-related proteins (LRPs),3, 4 lipolysis stimulated receptor,5 and one or more heparan sulfate proteoglycans.6, 7 The syndecans are type I transmembrane proteoglycans bearing up to three heparan sulfate chains and, in some family members, two chondroitin/dermatan sulfate chains.8 Genetic studies in which heparan sulfate assembly was selectively altered in mouse hepatocytes by Cre-loxP targeting of two sulfotransferases demonstrated the importance of the heparan sulfate chains in hepatic TRL clearance.9, 10 Furthermore, direct genetic evidence has been provided showing that syndecan-1 is the primary heparan sulfate proteoglycan mediating hepatic clearance of TRLs in mice.11 Its role in lipoprotein clearance in human hepatocytes is less clear, with some data suggesting that its primary role may be in binding of the remnants to the cell surface with subsequent transfer to other receptors.12, 13
In many cultured cells, syndecan-1 is constitutively shed from the cell surface into the growth medium.14 Inducers such as growth factors, bacterial virulence factors, ceramide, and the protein kinase C agonist phorbol myristic acid (PMA; phorbol-12-myristate-13-acetate) accelerate shedding by activation of one or more secreted or membrane-associated metalloproteinases including matrix metalloproteinase 7 (MMP7), MMP9, MMP14, and a disintegrin and metalloproteinase 17 (ADAM17).14 Although the exact biological significance of syndecan-1 shedding is unclear in most cells, one important role appears to be in the regulation of chemokine activity during bacterial infection.15-17 The role of syndecan-1 shedding in lipoprotein metabolism has not been studied.
Here, we examined the activity of syndecan-1 in TRL clearance in Hep3B human hepatoma cells and in primary human hepatocytes. We show that syndecan-1 is expressed by both human hepatoma cells and primary human hepatocytes and demonstrate its action as a TRL receptor. Syndecan-1 undergoes spontaneous proteolytic shedding from both human and mouse hepatocytes mediated by ADAM17. In mice, lipopolysaccharide (LPS) administration induces syndecan-1 shedding from the liver and reduces hepatic VLDL catabolism. These findings suggest that shedding of syndecan-1 may play a role in plasma lipid homeostasis.
ADAM, a disintegrin and metalloproteinase; BIM I, bisindolylmaleimide I; DiD, 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindodicarbocyanine perchlorate; ELISA, enzyme-linked immunosorbent assay; HSPG, heparan sulfate proteoglycan; LDLR, low density lipoprotein receptor; LPS, lipopolysaccharide; LRP, LDLR-related protein; PMA, phorbol myristic acid; siRNA, small interfering RNA; TNF-α, tumor necrosis factor-α; TRL, triglyceride-rich lipoprotein; VLDL, very low density lipoprotein.
Materials and Methods
Mice and Animal Husbandry.
Wild-type C57BL/6 and Ldlr−/− mice on a C57BL/6 background were purchased from The Jackson Laboratory. All animals were housed and bred in vivaria approved by the Association for Assessment and Accreditation of Laboratory Animal Care located in the School of Medicine, University of California San Diego (UCSD), following standards and procedures approved by the UCSD Institutional Animal Care and Use Committee. Mice were maintained on a 12-hour-light/12-hour-dark cycle and provided ad libitum with water and standard rodent chow (Harlan Teklad).
Hep3B hepatocarcinoma cells were obtained from the American Type Culture Collection (Manassas, VA) and cultured in minimum essential medium supplemented with 10% fetal bovine serum, nonessential amino acids, sodium pyruvate, penicillin, and streptomycin. Normal human hepatocytes were obtained through the Liver Tissue Cell Distribution System (Pittsburgh, PA) funded by NIH Contract N01-DK-7-0004/HHSN267200700004C. Primary mouse hepatocytes were isolated and cultured as described previously11 and used within 2 days of isolation.
To measure spontaneous syndecan-1 shedding, Hep3B cells were seeded in 12-well plates (3 × 105 cells) and incubated in serum-free medium. To induce shedding, cells were stimulated with 0.25 μM PMA (Sigma-Aldrich, St. Louis, MO) for 1 hour at 37°C. The effect of various metalloproteinase inhibitors and signaling pathway inhibitors was surveyed by pretreating cells for 1 hour with 10 μM Marimastat (Tocris bio, Ellisville, MO), GI254023 or GW280264 (provided by Dr. Andreas Ludwig, RWTH Aachen University, Aachen, Germany), bisindolylmaleimide I (BIM I), U0126 or SB 203580 (EMD, San Diego, CA), or vehicle only (dimethylsulfoxide; Sigma-Aldrich). In all shedding assays, conditioned media were collected and centrifuged to remove cell debris before further analysis.
Conditioned media or diluted plasma samples were applied to cationic polyvinylidene fluoride–based membranes (Hybond-N+; Amersham Biosciences, Piscataway, NJ) under mild vacuum in a bio-dot apparatus (Bio-Rad Laboratories, Hercules, CA). The membrane was blocked in 5% nonfat milk buffer and incubated with human syndecan-1 monoclonal antibody (mAb) B-A38 (AbD Serotec, Raleigh, NC) or mouse syndecan-1 mAb 281-2 (BD Biosciences, San Diego, CA) and a secondary horseradish peroxidase–conjugated anti-mouse immunoglobulin G (IgG; BD Bioscience, Franklin Lakes, NJ) or anti-rat IgG (Santa Cruz Biotechnology, Santa Cruz, CA). Blots were visualized by chemiluminescent substrate (Pierce, Rockford, IL) and analyzed with an Alpha Innotech Imager system (Cell Biosciences, Santa Clara, CA). All values were scaled relative to that obtained for the indicated control in each figure and expressed as a percentage.
Hepatocytes were incubated with a mixture of 2 mU/mL of heparin lyase I and II and 5 mU/mL of heparin lyase III in serum-free medium at 37°C for 15 minutes. Cells were subsequently lysed in radioimmunoprecipitation assay buffer supplemented with 1× protease inhibitor cocktail (Sigma-Aldrich). Each sample (10 μg protein) was resolved on a 4%-12% Bis-Tris NuPage gel (Invitrogen) and transferred to polyvinylidene fluoride membrane (Bio-Rad Laboratories). The membrane was blocked with Super-Block buffer (Pierce). Blots were incubated with B-A38 (1:500) or anti-ADAM17 polyclonal antibody ab2051 (1:500) (Abcam, Cambridge, MA) and secondary antibodies (horseradish peroxidase–conjugated anti-mouse IgG; BD Biosciences). Reactive bands were visualized by chemiluminescence.
Small interfering RNAs (siRNAs) specifically targeting human ADAM17 (5′-AGAGAGUACAACUACAAA-3′, 5′-GCAGUAAACAAUCAAUCU-3′, and 5′-GGAGAUUUGUUAAUGAUA-3′) and human syndecan-1(5′-AGAUAUCACCUUGUCACA-3′, 5′-CCAGUAGACCUUGUUACU-3′, and 5′-GGAGACAGCAUCAGGGUU-3′) were obtained from Integrated DNA Technologies (Coralville, IA). A scrambled siRNA was used as a control. All siRNAs were transfected into cells by using Transfectin (Integrated DNA Technologies, San Diego, CA).
VLDL Binding and Uptake.
Human VLDL (density δ < 1.006 g/mL) was isolated from plasma by buoyant density ultracentrifugation as described11 and quantified by BCA protein assay (Pierce). To label the particles, 1-2 mg of VLDL were combined with 100 μL of 3 mg/mL 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindodicarbocyanine perchlorate (DiD; Invitrogen) in dimethylsufoxide and then reisolated by ultracentrifugation. After incubation with VLDL, cells were rinsed with phosphate-buffered saline and lysed by adding 0.1 M NaOH plus 0.1% sodium dodecyl sulfate for 40 minutes at room temperature.18 Fluorescence intensity was measured with appropriate excitation and emission filters in a plate reader (TECAN GENios Pro; Tecan, Switzerland) and normalized to total cell protein. In some experiments, VLDL binding and uptake was measured by flow cytometry.
VLDL Plasma Clearance.
Mice were fasted for 6 hours and injected via the tail vein with 20 μg human VLDL protein. Serial samples were taken by tail vein bleeds, and the amount of human VLDL remaining in the plasma was determined by sandwich enzyme-linked immunosorbent assay (ELISA) as previously described and plotted relative to VLDL detected at 2 minutes.19
To radiolabel glycosaminoglycans, hepatoma cells were incubated for 48 hours in growth medium supplemented with 10% dialyzed fetal bovine serum containing 100 μCi/mL Na[35S]O4 (PerkinElmer Life Science, Waltham, MA). Shed [35S]syndecan-1 ectodomains were isolated from conditioned medium after incubation of cells with PMA. After 5 hours, proteoglycans were purified from the conditioned medium by anion exchange chromatography as described.20 Approximately 125 ng syndecan-1 ectodomains (determined by Sdc-1 ELISA) were diluted in 200 μL of a solution of iodixanol (δ = 1.019 g/mL; OptiPrep Density Gradient Medium; Sigma-Aldrich) at the bottom of an ultracentrifuge tube in the presence or absence of 50 μg VLDL with and without 50 μg heparin. The tubes were then carefully filled with a solution of iodixanol (δ = 1.019 g/mL) and centrifuged at 45,000 rpm (250,000g) for 3 hours at 18°C in a Beckman 50.4 Ti rotor. Fractions were removed sequentially from the top of the tubes and assayed for radioactivity by liquid scintillation counting.
Syndecan-1 Shedding In Vivo.
C57Bl/6 mice, 8-10 weeks old, were injected intraperitoneally with 4.5 mg/kg E. coli LPS. Eighteen hours later, food was removed from the cages. After 6 hours of fasting, the animals were sacrificed, and their livers and blood were harvested. Syndecan-1 in plasma samples was measured by dot blotting. Syndecan-1 in liver sections was visualized using mAb 281-2 as described.11 Plasma triglyceride was measured using Triglyceride-SL Reagent (Genzyme Diagnostics, Framingham, MA) and cholesterol via Roche Cobas C111 analyzer (Roche Diagnostics, Indianapolis, IN).
All data were analyzed by unpaired one-tailed t tests. P < 0.05 was considered statistically significant.
Human Hepatocytes Bind VLDL in a Heparan Sulfate-Dependent Manner.
Binding of DiD-labeled VLDL (DiD-VLDL) at 4°C to Hep3B human hepatoma cells occurred in a saturable manner both with respect to time and concentration (Fig. 1A,C, respectively). Treatment of the cells with a mixture of heparin lyases reduced binding by 80%-90%, indicating that the majority of binding sites were attributable to heparan sulfate proteoglycans on the cell surface. Analysis of the binding curve in Fig. 1C by a conventional single-site binding equation yielded a maximum binding (Bmax) value of 69 ± 12 μg VLDL/mg cell protein and a dissociation constant (KD) of 8.8 ± 3.4 μg/mL (R2 = 0.9980). Assuming that approximately 1.8% of the mass of VLDL particles is protein and an average molecular mass for VLDL of ∼2 × 107 Da,21 we calculated that Hep3B cells express ∼7 × 105 binding sites/cell attributable to heparan sulfate, similar to previous studies of mouse hepatocytes.11 Incubation of cells at 37°C increased the association of DiD-VLDL (Fig. 1B,D) because of internalization of the particles, as reported.22 Inclusion of heparin (100 μg/mL) blocked binding and uptake five-fold as measured by flow cytometry (data not shown).
Human hepatocytes express multiple heparan sulfate proteoglycans, including the membrane proteoglycans syndecan-1, syndecan-2, and syndecan-4 and glypican-1 and glypican-4, and the extracellular matrix proteoglycans perlecan, collagen 18, and agrin23, 24 (Supporting Fig. 1). To examine the contribution of syndecan-1 to VLDL binding and uptake, we silenced its expression on Hep3B cells using specific siRNAs (Supporting Fig. 2). DiD-VLDL binding at 4°C (Fig. 1E) and uptake at 37°C (Fig. 1F) was reduced compared to cells treated with a scrambled siRNA. Binding and uptake were diminished to a greater extent by heparan lyase digestion (Fig. 1E,F), suggesting that either the extent of syndecan-1 silencing was incomplete or that other heparan sulfate proteoglycans can mediate binding and uptake.6 Silencing of syndecan-4 did not result in reduction of binding, but other proteoglycans have not been examined (data not shown). The residual heparin lyase-insensitive component of binding/uptake presumably reflects other receptors, most likely LDL receptors and one or more members of the family of LDLR-related proteins.3, 4
Hepatocytes Shed Syndecan-1.
Syndecan-1 undergoes proteolytic shedding in many cells,14 resulting in the appearance in the medium of the extracellular domains of the protein containing the ligand binding heparan sulfate chains. To investigate whether human hepatocytes spontaneously shed syndecan-1, we collected conditioned growth media from Hep3B cells (Fig. 2A) and primary human hepatocytes (Fig. 2B) and measured syndecan-1 ectodomains by dot blotting. Syndecan-1 ectodomains accumulated progressively in the conditioned media over time in both types of cells. The broad-spectrum metalloproteinase inhibitor Marimastat blocked spontaneous shedding of syndecan-1 (Fig. 2A,B), consistent with the idea that shedding depends on limited proteolysis.
Syndecan-1 shedding can be induced in cells by PMA.14 In both Hep3B cells and primary human hepatocytes, PMA induced syndecan-1 shedding by ∼10-fold (Fig. 2C,D). Induction of shedding was dependent on both time of incubation and the concentration of PMA added to the medium (Supporting Fig. 3). Syndecan-1 on the cell surface was diminished in a corresponding manner, consistent with shedding (Fig. 2E,F). PMA is known to stimulate endocytosis, which could account for some reduction in cell surface expression of syndecan-1 as well.25 The protein kinase C inhibitor BIM I profoundly inhibited syndecan-1 ectodomain accumulation in conditioned media and prevented loss of syndecan-1 from the cell surface (Fig. 3A and Supporting Fig. 4A). In contrast, the MEK (MAPK/ERK kinase) inhibitor U0216 and the p38 mitogen-activated protein kinase (MAPK) inhibitor SB203580 failed to inhibit PMA-induced shedding of syndecan-1.
Marimastat blocked syndecan-1 shedding induced by PMA and spontaneous shedding (Fig. 2). To determine the relevant metalloproteinase(s), we tested two enzyme inhibitors on Hep3B cells. GW280264, which inhibits both ADAM17 and ADAM10, profoundly diminished the accumulation of syndecan-1 ectodomain in conditioned media (Fig. 3B) and prevented the PMA-induced loss of syndecan-1 from the cell surface (Supporting Fig. 4B). In contrast, the inhibitor GI254023, which preferentially blocks ADAM10 but not ADAM17, had no effect on PMA-induced shedding of syndecan-1. Silencing of ADAM17 expression in Hep3B cells by siRNA reduced ADAM17 expression approximately six-fold and prevented PMA-induced accumulation of syndecan-1 ectodomains in the conditioned medium (Fig. 3C) and prevented loss of syndecan-1 from the cell surface as compared to controls in which cells were treated with scrambled siRNA (Supporting Fig. 4C). Similar results were obtained with primary human hepatocytes (Fig. 4). These findings indicate that ADAM17 is the primary metalloproteinase responsible for PMA-induced syndecan-1 shedding in human hepatocytes.
Syndecan-1 Shedding Reduces VLDL Binding and Uptake.
In view of the role for syndecan-1 in TRL binding and uptake (Figs. 1 and 2),11 we predicted that PMA-induced syndecan-1 shedding would affect VLDL binding and uptake. In Hep3B cells, PMA reduced VLDL binding and uptake by ∼50% compared to untreated cells (Fig. 5A,B). Treatment with heparin lyases reduced VLDL binding and uptake to a greater extent, most likely because of incomplete removal of proteoglycan receptors from the cell surface by PMA-induced shedding. Similar results were obtained in primary human hepatocytes (Fig. 5C,D). Shed ectodomains did not inhibit binding when purified and added to fresh cells at their original concentration. PMA can also induce shedding of LRP-1, another TRL receptor, which might account for some decrease in binding.3, 26 However, the major remnant receptor appears to bear heparan sulfate, on the basis of the large inhibitory effect of heparin lyase treatment (Fig. 1).
Although the ADAM17 cleavage site in syndecan-1 is unknown,27 it presumably lies in the membrane proximal region, thus liberating the ectodomain with covalently attached heparan sulfate chains. Based on previous genetic studies of mouse syndecan-1,11 the heparan sulfate chains make up the TRL-binding domain of the syndecan-1 receptors. Thus, we predicted that shed syndecan-1 ectodomains from hepatocytes would retain the capacity to bind VLDL. To test this hypothesis, we incubated hepatoma cells with [35S]O4 to radiolabel the heparan sulfate chains on syndecan-1, triggered shedding with PMA, and purified the [35S]-labeled ectodomains from the medium by anion exchange chromatography. The [35S]-labeled ectodomains were then mixed with VLDL, placed at the bottom of a centrifuge tubes, and overlaid with a solution of iodixanol (δ = 1.019 g/mL). Ultracentrifugation resulted in the appearance of ∼65% of counts in the top four fractions, whereas in the absence of VLDL, less than 5% of the counts were found in the top fractions (Fig. 6). Dot blot analysis of the pooled top four fractions showed syndecan-1 ectodomains were present in samples containing VLDL (Fig. 6, inset). Inclusion of 50 μg of heparin in the binding solution reduced the recovery of [35S]-labeled ectodomains in the top fractions, consistent with the idea that heparin prevented the association of the particles with the heparan sulfate chains on the ectodomains. Interestingly, when the mixture of VLDL and [35S]-labeled ectodomains was overlaid with a lower density solution (δ = 1.006 g/mL), flotation of [35S]-ectodomains did not occur (data not shown), consistent with the idea that the association of ectodomains with VLDL caused a shift in the buoyant density of the complex.
LPS Causes Syndecan-1 Shedding in Mice and Increases Plasma Triglycerides.
On the basis of these findings, we predicted that the steady-state level of plasma triglycerides might depend on the extent of syndecan-1 shedding that occurs in the liver. To examine this possibility, we isolated hepatocytes from wild-type C57BL/6 mice and examined syndecan-1 shedding. Syndecan-1 was constitutively shed from the cells based on the appearance of ectodomains in the conditioned media (Fig. 7A). However, in contrast to human hepatocytes, shedding was insensitive to Marimastat (or PMA stimulation; data not shown). Other factors previously shown to induce shedding in other cells types such as insulin, LPS, and tumor necrosis factor-α (TNF-α), also were without effect.28-30
Hayashida et al. recently showed that in mice, injection of LPS induces syndecan-1 shedding in liver.16 We confirmed this finding by dot blotting of plasma samples 24 hours after injection of 4.5 mg LPS/kg body weight into mice, compared to the level in mice injected with vehicle (Fig. 7B). LPS injection also resulted in dramatic loss of syndecan-1 in the liver, as measured by immunohistochemistry (Fig. 7C). Analysis of fasting plasma lipids showed that triglyceride levels increased two-fold in mice injected with LPS (Fig. 8A), whereas plasma total cholesterol did not change (Fig. 8B). The accumulation of plasma lipids was greatly accentuated under these conditions by deletion of the LDL receptor, which resembles the phenotype of compound mutants lacking hepatic heparan sulfate and LDL receptors (Supporting Fig. 5).9 When the animals were challenged with human VLDL, the injected particles cleared less extensively in the LPS-injected mice (Fig. 8C; area under the curve = 9200 ± 400 for LPS-injected mice versus 7000 ± 400 for the control), consistent with a loss of syndecan-1 clearance receptors in the liver.
Here, we show that the heparan sulfate proteoglycan syndecan-1 mediates clearance of VLDL in human hepatoma cells and primary human hepatocytes. Furthermore, we demonstrate that shed syndecan-1 ectodomains from human hepatoma cells bind VLDL particles in vitro (Fig. 6), consistent with the idea that ectodomains contain the ligand binding site of the syndecan-1 receptor. Importantly, stimulation of shedding of syndecan-1 in mice was accompanied by an increase in fasting triglycerides (Fig. 8). Proteoglycan receptors appear to dominate the receptor landscape on human hepatocytes as on murine hepatocytes, representing at least 90% of the binding sites.11
Earlier studies of TRL uptake in human hepatocytes focused on hepatoma cell lines, such as HepG2 and Hep3B, that are derived from well-differentiated human hepatocellular carcinomas12, 31, 32 or more rarely on primary hepatocytes.33 The data demonstrated that the addition of apolipoprotein E and lipoprotein lipase to particles enhanced their uptake, and that under these conditions, uptake occurred in a manner dependent on heparan sulfate. Similar observations were made using nonhepatic cell lines, which also suggested the participation of various syndecans34-39 and perlecan.34, 35, 37, 40 Zeng et al. showed that syndecan-1 can mediate binding and uptake of chylomicron remnants by HepG2 liver cells in vitro based on antisense and antibody inhibition experiments.12 In this report, we show that syndecan-1 on primary human hepatocytes as well as Hep3B cells can bind and take up native TRL particles derived from fasted donors. Prior genetic studies in mice and silencing experiments in human hepatocytes suggest that other heparan sulfate proteoglycans probably do not fulfill this role, emphasizing that syndecan-1 may be the dominant proteoglycan clearance receptor in humans as well as in mice.11
It has been estimated that as much as 4% of all cell-surface receptors undergo regulated proteolytic shedding.41 Syndecan-1 also undergoes proteolytic processing, resulting in shedding of ectodomains containing the attached glycosaminoglycan chains.42 The shedding process occurs in hepatocytes and appears to be mediated by ADAM17 both under basal conditions (data not shown) and after PMA stimulation (Figs. 3 and 4). The inducibility of syndecan-1 shedding by inflammation, ischemia, glucose, and insulin suggests that shed ectodomains might have functional significance.28, 43-46 Shed syndecan-1 ectodomains might bind plasma lipoproteins in the space of Disse and prevent their escape back into the plasma or facilitate their further processing prior to uptake. Because shedding of ectodomains increases plasma triglycerides (Fig. 8), shedding would appear to serve an alternative role, e.g., increasing the circulatory half-life of TRLs for more complete use of triglycerides in peripheral tissues. In order to study this question, studies are underway that use recombinant forms of syndecan-1 that fail to shed or which are shed constitutively.
Shedding of syndecan-1 in hepatocytes depends on ADAM17, which can be induced by treatment with PMA through a pathway dependent on protein kinase C. ADAM17, also known as TNF-α–converting enzyme, processes membrane-bound pro–TNF-α, releasing the soluble active form of TNF-α, as well as other factors involved in inflammation, including L-selectin,46-48 LRP1,43 TNF-α receptors TNFR1 and TNFR2,49 CD30,50 and interleukin-6 receptor IL-6R.51 Park and colleagues have demonstrated that the role of syndecan-1 ectodomains in lung inflammation is to modulate the availability of chemokines.52 It is also well known that LPS can bind to lipoproteins, which serves as a sink for this potent inflammatory mediator.53 Thus, one can imagine that activation of shedding and the ensuing increase in plasma TRLs might serve a protective role during infection. The observation that LPS induces shedding, elevates plasma TRLs, and reduces VLDL clearance is consistent with this hypothesis and suggests that syndecan-1 shedding might be responsible in part for the elevation of plasma triglycerides in bacterial sepsis.54, 55 Further studies are planned to determine whether syndecan-1 shedding contributes to the hyperlipidemia of sepsis in human patients and whether it might explain other idiopathic forms of hypertriglyceridemia, a common side effect of many drugs.56
We thank Joe Juliano for performing lipid analyses, Andrea Garcia and Dr. Nissi Varki for their help on the syndecan-1 immunohistochemistry, and Drs. Pyong Park and Joseph Witztum for many helpful discussions.