LpS treatment. To induce inflammatory response, mice were treated with LPS diluted in sterile saline and administered intraperitoneally (IP) once a day for three days (1 mg kg −1 bw-IP). Mice were euthanized 2 hours after LPS treatment, and fragments of liver were collected, froze in liquid nitrogen and stored at −80 °C until processing. In vitro experiments. Hepatoma cell line, Hepa-1c1c7 (ATCC ® CRL-2026 ™), derived from mice was used to evaluate the ability of the cholinergic pathway to modulate AKT phosphorylation induced by insulin. Cells were cultivated in alpha modified Eagle's medium (αMEM; Invitrogen, USA) supplemented with 10% foetal bovine serum (Invitrogen, USA) and 1% penicillin (100 U/mL)/streptomycin (100 µg/mL) (Invitrogen, USA) at 37 °C and 5% CO 2. Cells were treated with 500 µM palmitate (palmitic acid from Sigma-Aldrich at 500 μM was first diluted in NaOH conjugated to BSA (3:1) for 45 minutes at 37 °C) for 3 hours in 6-well culture plate. The protein content was extracted and analysed by Western blotting. When necessary, 1 µM PNU-282987 (P6499-10MG; Sigma-Aldrich, Brazil) or 1 µM nicotine (N0267-100MG; Sigma-Aldrich, Brazil) was added to the medium for 15 minutes after palmitate treatment. To evaluate the insulin signalling, cells were treated with 100 nM insulin (Humulin, Eli Lilly and Company, USA) for 10 minutes after the 3 hours of palmitate treatment.

Activation of cholinergic receptor (α7nAChR) in hypothalamus can modulates inflammatory response and they are colocalized with POMC and NPY neurons. Activation of hypothalamic nAChR modulated NPY and POMC expression and reduced food intake. Maternal obesity promoted reduction of nAChR in hypothalamus of the offspring and increased food intake. Thus, the reduction of hypothalamic α7nAChR in offspring may be related to obesity development induced by maternal consumption of high fat diet during pregnancy and lactation.

Studies has shown that the consumption of a high-fat diet induces hypothalamic and systemic inflammation. It is also known that nicotinic acetylcholine receptors (nAChRs) are involved in cholinergic anti-inflammatory response, in which the α7-subunit is the most expressive subtype doing this control in immune cells. This study investigated the effects of a short-term high-fat diet on modulating the anti-inflammatory pathway mediated by α7 receptor. 8-weeks-old Swiss mice consumed either short-term (3 days) high-fat diet (HFD-60% of fatty acids) or standard-chow. Both groups were submitted to either a CLP surgery, to induce polymicrobial sepsis, or an abdominal incision for exposure of the cecum, serving as the control group. We concluded that the consumption of a high-fat diet even for a short period is already capable of increasing hypothalamic anti-inflammatory markers and inhibiting the anti-inflammatory pathway .

The insulin resistance is especially caused by inflammatory process in obesity. The activation of antiinflammatory cholinergic pathway is responsible to attenuatte inflammation. The objective was investigate if the obese offspring have cholinergic antiinflammatory signalling damage and insulin resistance impaired. The inflammation LPS-induced was able to increase pIKK, IL-6, pSTAT3 levels and decrease gluconeogenesis in lean and obese offspring. The treatment seems to reduce α7nAChR in offspring and increase to pAKT in obese offspring. Our datas indicate which the maternal obesity impaired the cholinergic signalling and this can be affect glucose homeostasis through IL-6 signalling.

Gestational obesity is one of the most common obstetric risks and its prevalence has been increasing substantially. It is known that in obesity there is a loss of the cholinergic signalling responsible for the anti-inflammatory reflex which leads to an immune system imbalance. We hypothesize that the offspring (28 days old) of the maternal high fat diet consumption during the gestational and the lactation periods shows an impared cholinergic anti-inflammatory reflex. From this point on, an increase of inflammatory cytokines in the HFD-O spleen, such as TNF and IL1β, was observed, and this increase seems to be responsive to the smallest phosphorylation of STAT3 and the increase of the expression of NFkB.