TLR4

Lipopolysaccharides (10–80 μg/mL) selectively reduce the production of nitric oxide (NO) in THir (tyrosine hydroxylase immunoreactive) cells and increase the levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) as well as nitrite (nitrate index) in the culture medium [5].

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Bacterial lipopolysaccharides (LPS) are unique and complex glycolipids that provide characteristic components of the outer membranes of Gram-negative bacteria. In LPS of the Enterobacteriaceae, the core oligosaccharide links a highly conserved lipid A to the antigenic O-polysaccharide. Structural diversity in the core oligosaccharide is limited by the constraints imposed by its essential role in outer membrane stability and provides a contrast to the hypervariable O-antigen. The genetics of core oligosaccharide biosynthesis in Salmonella and Escherichia coli K-12 have served as prototypes for studies on the LPS and lipo-oligosaccharides from a growing range of bacteria. However, despite the wealth of knowledge, there remains a number of unanswered questions, and direct experimental data are not yet available to define the precise mechanism of action of many gene products. Here we present a comparative analysis of the recently completed sequences of the major core oligosaccharide biosynthesis gene clusters from the five known core types in E. coli and the Ra core type of Salmonella enterica serovar Typhimurium and discuss advances in the understanding of the related biosynthetic pathways. Differences in these clusters reflect important structural variations in the outer core oligosaccharides and provide a basis for ascribing functions to the genes in these model clusters, whereas highly conserved regions within these clusters suggest a critical and unalterable function for the inner region of the core[3].

Induction of cardiac dysfunction [8] Pathogenic principle Cardiac dysfunction (cardiac inflammation) is a common complication of lipopolysaccharide (LPS)-induced sepsis and is caused by a severe inflammatory response. LPS can also cause deterioration of cardiac systolic function and increase cardiac lipid peroxidation, leading to cardiac dysfunction. Specific modeling method Rat[8]: male • Wistar male rats • 6-week-old Administration: 10 mg/kg • ip for single dose • collected cardiac tissues and serums 24 hr after LPS-injection. Note / Modeling success indicators Phenotypic changes: left ventricular end-systolic volume (LVESV, %) and left ventricular end-diastolic volume (LVEDV, %) of cardiac function were significantly increased. Molecular level: CK-MB, LDH, and AST levels in serum are ↑, and the TLR4/NF-κB pathway is inhibited. Antagonist product Ferrostatin-1[7].

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Puberty is an important developmental event that is marked by the reorganizing and remodeling of the brain. Exposure to stress during this critical period of development can have enduring effects on both reproductive and non-reproductive behaviors. The purpose of this study was to investigate age and sex differences in immune response by examining sickness behavior, body temperature changes, and serum cytokine levels following an immune challenge. The effects of circulating gonadal hormones on age and sex differences in immune response were also examined. Results showed that male mice display more sickness behavior and greater fluctuations in body temperature following LPS treatment than female mice. Moreover, adult male mice display more sickness behavior and a greater drop in body temperature following LPS treatment compared to pubertal male mice. Following gonadectomy, pubertal and adult males displayed steeper and prolonged drops in body temperature compared to sham-operated counterparts. Gonadectomy did not eliminate sex differences in LPS-induced body temperature changes, suggesting that additional factors contribute to the observed differences. LPS treatment increased cytokine levels in all mice. However, the increase in pro-inflammatory cytokines was higher in adult compared to pubertal mice, while the increase in anti-inflammatory cytokines was greater in pubertal than in adult mice. Our findings contribute to a better understanding of age and sex differences in acute immune response following LPS treatment and possible mechanisms involved in the enduring alterations in behavior and brain function following pubertal exposure to LPS[2].

Human podocyte cell line (HPC) was used. HPCs were seeded onto culture plates and cultured in RPMI 1640 medium enriched with 10% FBS, 100 U/mL penicillin and 100 μg/mL streptomycin, and ITS. HPCs were cultured at 33°C and 5% CO2 for proliferation and were then shifted to 37°C and 5% CO2 for differentiation for 10–12 days. HPCs were incubated with or without different dose LPS, PAN, and HG for different time. HPCs were cultured with or without 50 μg/mL LPS, 75 μg/mL PAN, 60 mM HG, and different dose Rac-1 inhibitor (EHT 1864) for different time. In addition, HPCs were cultured with dynamin Inhibitor Dynasore or blebbistatin for 12 h[3].

Animal/Disease Models:Female and male CD1 mice[3]
Doses: 1.5mg/kg
Route of Administration: Intraperitoneal injection, once
Experimental Results: Induced sickness behavior in all mice, but adult mice displayed more sickness than pubertal mice and adult males remained sick for a longer period of time than adult females. Caused a decrease in body temperature for all mice, but this decrease was greatest in adult males. Increased pro- and anti-inflammatory cytokines at various levels in pubertal and adult male and female mice, resulted in age and sex differences in cytokine concentrations following immune challenge. Only adult males and females treated with LPS displayed significantly more IL-6 than their saline controls, and pubertal males and females and adult females displayed significantly more IL-10 than their saline controls. All the mice displayed significantly more IL-12 and TNF-α than their saline controls.

[1]. Structural analysis of lipopolysaccharides from Gram-negative bacteria. Biochemistry (Mosc). 2010 Apr;75(4):383-404.

[2]. Age and sex differences in immune response following LPS treatment in mice. Brain Behav Immun. 2016 Nov;58:327-337.

[3]. Molecular basis for structural diversity in the core regions of the lipopolysaccharides of Escherichia coli and Salmonella enterica. Mol Microbiol. 1998 Oct;30(2):221-32.

[4]. Podocyte-Released Migrasomes in Urine Serve as an Indicator for Early Podocyte Injury. Kidney Dis (Basel). 2020 Nov;6(6):422-433.

[5]. Lipopolysaccharide (LPS)-induced dopamine cell loss in culture: roles of tumor necrosis factor-alpha, interleukin-1beta, and nitric oxide. Brain Res Dev Brain Res. 2002 Jan 31;133(1):27-35.

[6]. Ren Q, Guo F, Tao S, Huang R, Ma L, Fu P. Flavonoid fisetin alleviates kidney inflammation and apoptosis via inhibiting Src-mediated NF-κB p65 and MAPK signaling pathways in septic AKI mice. Biomed Pharmacother. 2020 Feb;122:109772.

[7]. Koc F, Tekeli MY, Kanbur M, Karayigit MÖ, Liman BC. The effects of chrysin on lipopolysaccharide-induced sepsis in rats. J Food Biochem. 2020 Sep;44(9):e13359.

[8]. Xiao Z, Kong B, Fang J, Qin T, Dai C, Shuai W, Huang H. Ferrostatin-1 alleviates lipopolysaccharide-induced cardiac dysfunction. Bioengineered. 2021 Dec;12(2):9367-9376.

Background: Levels of urinary microvesicles, which are increased during various kidney injuries, have diagnostic potential for renal diseases. However, the significance of urinary microvesicles as a renal disease indicator is dampened by the difficulty to ascertain their cell source. Objectives: The aim of this study was to demonstrate that podocytes can release migrasomes, a unique class of microvesicle with size ranging between 400 and 2,000 nm, and the urine level of migrasomes may serve as novel non-invasive biomarker for early podocyte injury. Method: In this study, immunofluorescence labeling, electronic microscopy, nanosite, and sequential centrifugation were used to purify and analyze migrasomes. Results: Migrasomes released by podocytes differ from exosomes as they have different content and mechanism of release. Compared to podocytes, renal tubular cells secrete markedly less migrasomes. Moreover, secretion of migrasomes by human or murine podocytes was strongly augmented during podocyte injuries induced by LPS, puromycin amino nucleoside (PAN), or a high concentration of glucose (HG). LPS, PAN, or HG-induced podocyte migrasome release, however, was blocked by Rac-1 inhibitor. Strikingly, a higher level of podocyte migrasomes in urine was detected in mice with PAN-nephropathy than in control mice. In fact, increased urinary migrasome number was detected earlier than elevated proteinuria during PAN-nephropathy, suggesting that urinary migrasomes are a more sensitive podocyte injury indicator than proteinuria. Increased urinary migrasome number was also detected in diabetic nephropathy patients with proteinuria level <5.5 g/day. Conclusions: Our findings reveal that podocytes release the "injury-related" migrasomes during migration and provide urinary podocyte migrasome as a potential diagnostic marker for early podocyte injury.[4]

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Parkinson's disease (PD) is a neurodegenerative disorder characterized by the loss of dopamine (DA) neurons of the substantia nigra pars compacta (SNc). Although the exact mechanisms responsible for this cell loss are unclear, emerging evidence suggests the involvement of inflammatory events. In the present study, we characterized the effects of the proinflammatory bacteriotoxin lipopolysaccharide (LPS) on the number of tyrosine hydroxylase immunoreactive (THir) cells (used as an index for DA neurons) in primary mesencephalic cultures. LPS (10-80 microg/ml) selectively decreased THir cells and increased culture media levels of interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) as well as nitrite (an index of nitric oxide (NO) production). Cultures exposed to both LPS and neutralizing antibodies to IL-1beta or TNF-alpha showed an attenuation of the LPS-induced THir cell loss by at least 50% in both cases. Inhibition of the inducible form of nitric oxide synthase (iNOS) by L-NIL did not affect LPS toxicity, but increased the LPS-induced levels of both TNF-alpha and IL-1beta. These findings suggest that neuroinflammatory stimuli which lead to elevations in cytokines may induce DA neuron cell loss in a NO-independent manner and contribute to PD pathogenesis.[5]

White to off-white solid powder

Lipopolysaccharide; LPS

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : 14.29 mg/mL (with sonication (<60°C))
H2O : 5 mg/mL

Solubility in Formulation 1: ≥ 1.43 mg/mL 1 (saturation unknown) in 10% DMSO 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one). For example, to prepare 1 mL of working solution, add 100 μL of 14.3 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD in saline and mix. For the following dissolution protocols, prepare the working solution directly. It is recommended to prepare it as needed and use it up as soon as possible. The percentages shown in front of the following solvents refer to the volume percentage of the solvent in your final solution; if precipitation or precipitation occurs during the preparation process, heating and/or ultrasound can be used to assist dissolution.
* Preparation of 20% SBE-β-CD saline solution (4°C, 1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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 (Please use freshly prepared in vivo formulations for optimal results.)