Treatment of experimental non‑alcoholic steatohepatitis by targeting α7 nicotinic acetylcholine receptor‑mediated inflammatory responses in mice

  • Authors:
    • Zhou Zhou
    • Ying‑Chao Liu
    • Xiao‑Mei Chen
    • Fu‑Qiang Li
    • Xiao‑Juan Tong
    • Yue‑Ping Ding
    • Cui‑Lan Tang
  • View Affiliations

  • Published online on: September 10, 2015     https://doi.org/10.3892/mmr.2015.4318
  • Pages: 6925-6931
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

Non‑alcoholic fatty liver disease (NAFLD) is one of the most common types of liver disease, affecting up to 30% of the general population worldwide. Non‑alcoholic steatohepatitis (NASH) is a severe form of NAFLD without any effective therapies available. The present study showed that activation of α7‑nicotinic acetylcholine receptor (α7 nAChR) may be a novel potential strategy for NASH therapy. Treatment with the α7 nAChR agonist nicotine for three weeks obviously attenuated hepatic steatosis in a high-fat diet‑induced mouse model of NASH. Investigation of the underlying mechanism showed that nicotine reduced the secretion of the pro‑inflammatory cytokines tumor necrosis factor α and interleukin 6 in vitro and in vivo. Inflammation is an integral part of NASH and is the most prevalent form of hepatic pathology found in the general population; therefore, the effect of α7 nAChR activation against NASH may be ascribed to its anti‑inflammatory effects. In addition, the present study showed that nicotine‑stimulated α7 nAChR activation led to a significant downregulation of nuclear factor kappa B (NK‑κB) and extracellular signal-regulated kinase (ERK). It therefore appeared that activation of α7 nAChR suppressed the production of pro‑inflammatory cytokines through NK‑κB and ERK pathways. In conclusion, the present study indicated that targeting α7 nAChR may represent a novel treatment strategy for NASH.

Introduction

Non-alcoholic fatty liver disease (NAFLD) is a cause of fatty liver, occurring when fat is deposited in the liver (steatosis) not due to excessive alcohol use. It is associated with insulin resistance and metabolic syndrome (1). NAFLD is currently considered to be the most common cause of chronic liver disease worldwide (2) and associated with other potentially life-threatening diseases and increased mortality from cardiovascular diseases, malignancy and hepatic complications. NAFLD has also been found to be associated with several extra-hepatic disorders, including breast cancer, polycystic ovary syndrome and renal dysfunction (315). NAFLD encompasses a wide spectrum of liver diseases ranging from simple steatosis to non-alcoholic steatohepatitis (NASH) (1), which is the most extreme form of NAFLD and is regarded as a major cause of cirrhosis of the liver of unknown cause (16). NASH is a major health problem and complicated by portal hypertension and hepatic decompensation, and is occasionally accompanied with hepatocellular carcinoma (HCC) (17).

Recently, various treatment modalities have been applied in NASH, including lifestyle modification, surgical intervention and pharmacological agents (including insulin sensitizers, anti-oxidant agents, lipid-lowering agents and tumor necrosis factor-alpha (TNF-α) antagonists) (1820). However, to date, there are no US Food and Drug Administration-approved medical therapies for NASH or liver fibrosis. There is an urgent requirement for novel therapeutic approaches (17,21). As inflammatory activation has a significant role in NASH progression, anti-inflammatory therapy for NASH is of increasing interest (22). For example, TNF-α antagonist pentoxifylline, interleukin (IL)-6 antagonist Sant7 and the TNF-α-specific monoclonal antibodies infliximab, adalimumab and certolizumab have been studied in a number of clinical NAFH trials (23). At present, anti-inflammatory strategies for NASH are restricted to targeting one single cytokine, e.g., IL-1 receptor, IL-6 or TNF-α. However, multiple cytokines are involved in the inflammatory response of NASH. Therefore, targeting an upstream signaling molecule that regulates multiple cytokine production may improve the objective response rates (24). Recently, a novel neural pathway termed as cholinergic anti-inflammatory reflex, has been discovered, which inhibits the production of inflammatory cytokines and may be a novel anti-inflammatory strategy for NASH.

The α7-nicotinic acetylcholine receptor (α7 nAChR) is a sub-type of nicotinic acetylcholine receptor and has a crucial role in mediating the cholinergic anti-inflammatory signaling pathway (25). It is expressed on different types of cells, including neurons, macrophages, lymphocytes, monocytes and dendritic cells. Activation of the α7 nAChR expressed on resident macrophages may suppress the local inflammation by reducing the production of pro-inflammatory cytokines TNF-α and IL-6, which are closely associated with certain inflammatory diseases, including sepsis, rheumatoid arthritis, asthma and diabetes (26). It is therefore indicated that α7 nAChR is a promising target for developing novel anti-inflammatory drugs. However, to date, it has remained to be clarified whether α7 nAChR is associated with NASH.

The present study assessed whether activation of α7 nAChR was able to prevent the progression of NASH, and whether targeting of α7 nAChR may represent a novel strategy for NASH therapy.

Materials and methods

Cell culture and reagents

RAW 264.7 cells were obtained from the American Type Culture Collection (Manassas, VA, USA) and maintained in Dulbecco's modified Eagle's medium (Gibco-BRL, Invitrogen Life Technologies, Carlsbad, CA, USA) supplemented with 10% heat-inactivated fetal bovine serum (Invitrogen Life Technologies) in a humidified atmosphere of 95% air with 5% CO2 at 37°C. Nicotine was purchased from Sigma-Aldrich (St. Louis, MO, USA; n=80).

Experimental protocols and animals

C57 male mice at four weeks of age (weight, 17–23 g) were purchased from the Model Animal Research Center of Nanjing University (Nanjing, China) and housed in the laboratory animal center of Zhejiang Chinese Medical University (Hangzhou, China) at 22°C with a 12-h light/dark cycle. Mice were randomly divided into four groups (n=10) and fed either a control diet (10% kcal as fat; Mediscience Ltd., Yangzhou, China) or a high-fat diet (HFD; 60% kcal as fat; Medicience Ltd) for 18 weeks with or without nicotine for three weeks: 1) Control group, mice were fed a control diet and supplemented with normal saline; (2) HFD group, mice were fed a HFD and supplemented with normal saline; (3) control + nicotine 5 mg/kg group, mice were fed a control diet and supplemented with nicotine at a dose of 5 mg/kg; (4) HFD + nicotine 5 mg/kg group, mice were fed a HFD and supplemented with nicotine at the dose of 5 mg/kg. During the 18 weeks of feeding, the body weight was measured every week. At the end of the experiment, mice were sacrificed by cardiac puncture under CO2 anesthesia, and livers were collected for further analysis.

All animals used in the present study were housed and cared for in accordance with the Chinese Pharmacological Society Guidelines for Animal Use. The protocols of the present study were approved by the Committee on the Ethics of Animal Experiments of the Zhejiang Chinese Medical University (Hangzhou, China; permit no. 2012-1849). All surgeries were performed under sodium pentobarbital anesthesia (70 mg/kg; Sigma-Aldrich) and all efforts were made to minimize suffering.

Biochemical serum analysis

The activity levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) (10) were determined using an automatic blood chemical analyzer (Dry-Chem 4000i; Fujifilm, Tokyo, Japan). TNF-α and IL-6 levels were measured using ELISA kits (cat. nos. EK0527 and EK0441; Boster Biological Inc., Wuhan, China).

Histological examination and Oil Red O staining

The fixed liver tissue was cut into 3-mm blocks, which were embedded in paraffin and cut into 4-µm slices. After being de-paraffinized using xylene and ethanol dilutions and re-hydration, the sections were stained with hematoxylin and eosin (H&E; Bogoo, Shanghai, China) to examine the tissue structure, inflammatory cell infiltration, necrosis and lipid accumulation.

For Oil Red O staining, cryosections of optimal cutting temperature compound-embedded liver tissues (10 mm) were fixed in 10% buffered formalin for 5 min at room temperature, stained with Oil Red O (Biohao Company, Wuhan, China) for 1 h, washed with 10% isopropanol and then counterstained with hematoxylin for 30 sec. A Nikon E600 microscope (Nikon, Tokyo, Japan) and Leica Application Suite (Leica Microsystems, Inc., Buffalo Grove, IL, USA) were used to capture images of the Oil Red O-stained tissue sections at 40× magnification.

Isolation of macrophages from liver tissue

Forty normal, healthy mice were anesthetized and liver tissues were perfused in situ via the superior vena cava with a perfusion buffer (13 Hanks' balanced salt solution; Gino Biological Medical Technology, Co., Ltd., Hangzhou, China), followed by a digestion buffer [13 Hanks' balanced salt solution, supplemented with 0.05% collagenase (Type IV; Sigma-Aldrich), 1.25 mmol/l CaCl2, 4 mmol/l MgSO4 and 10 mmol/l 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid]. The resulting cell suspension was filtered through a sterile 100-mm nylon mesh (Solarbio, Beijing, China) and centrifuged at 50 ×g to selectively sediment hepatocytes from non-parenchymal cells (NPCs). The pellet of hepatocytes was re-suspended and subsequently washed two more times with centrifugation at 50 ×g. The NPCs in the first and second supernatants from the low-speed centrifugations were pelleted by high-speed centrifugation (1,300 × g), followed by re-suspension in a small volume prior to isopycnic sedimentation in Percoll as previously described (27). Cell viability (90%) was determined by trypan blue exclusion (Sigma-Aldrich). The Kupffer cells were treated with lipopolysaccharide (LPS; Escherichia coli O111:B4; Sigma-Aldrich; 100 nM) for 16–18 h, following which the culture medium was replaced with medium without serum, and in the presence or absence of nicotine (concentrations between 0 and 10 µm) for 6 h. Treatment with α-bungarotoxin (α-BGT; Zhongxin Dongtai Company, Laiyang, China) was also performed.

Western blot analysis

Whole-cell lysates were prepared using radioimmunoprecipitation assay lysis buffer (Beyotime Institute of Biotechnology, Nantong, China), protein concentrations were detected using a bicinchoninic acid assay kit (Beyotime Institute of Biotechnology) and western blotting was performed, as previously described (27). Briefly, equal amounts of protein were separated by SDS-PAGE. Proteins were then transferred onto nitrocellulose membranes and identified with anti-α7 nAChR polyclonal antibody (cat. no. 23791-AP; Proteinch USA), anti-NF-κB monoclonal antibody (cat. no. 4764S; Cell Signaling Technology, Inc., Beverly, MA, USA), anti-inhibitor of NF-κB (IκB) antibody (cat. no. 4814; CST Company, Boston, MA, USA), anti-extracellular signal-regulated kinase (ERK) monoclonal antibody (cat. no. 20G11; Cell Signaling Technology, Inc.) and anti-GAPDH antibodies (Santa Cruz Biotechnology, Inc., Dallas, TX, USA) at 1:1,000. Detection was performed using a horseradish peroxidase-conjugated secondary antibody and SuperSignal West Pico Chemiluminescent Substrate (Pierce Biotechnology, Inc., Rockford, IL, USA) according to the manufacturer's instructions. Kodak films (Kodak, Rochester, NY, USA) were used to visualize the gels.

Statistical analysis

Values are expressed as the mean ± standard deviation. Statistical analyses were performed using one-way analysis of variance or the unpaired Student's t-test as indicated. Statistical analysis was performed using SPSS v.10.0 statistical software (SPSS, Inc., Chicago, IL, USA). P<0.05 was considered to indicate a statistically significant difference between values.

Results

HFD-induced NASH

In the present study, a mouse model of NASH was generated by intake of a HFD. After 18 weeks of HFD intake, the body weight was significantly increased, which indicated the establishment of the obesity mouse model (Fig. 1A). As shown in Fig. 2B and C, activities of AST and ALT were increased in mice on an HFD compared with those in the control mice which received a normal diet. It appeared that the HFD induced liver injury. To determine whether HFD induced hepatic steatosis, liver pathological examination by H&E staining was performed (Fig. 1D). The hepatic cell structure in the control group was normal. However, the HFD increased hepatic damage with obvious hepatic necrosis. Further examination of the hepatic lipid accumulation status with Oil Red O staining revealed that the HFD significantly induced hepatic lipid accumulation compared to that in the control group.

Activation of α7 nAChR attenuates HFD-induced hepatic steatosis

In order to identify whether activation of α7 nAChR can prevent NASH and the subsequent hepatic injury, α7 nAChR agonist nicotine was administered to mice receiving the HFD. As shown in Fig. 2, administration of nicotine significantly, but not completely, prevented HFD-induced hepatic necrosis and hepatic lipid accumulation.

Activation of α7 nAChR attenuates HFD-induced hepatic inflammation

Inflammation is the main pathological consequence of HFD-induced NASH and is characterized by a release of inflammatory factors, which contributes to hepatic fibrosis (28,29). Thus, the present study determined whether nicotine can prevent HFD-induced hepatic inflammation. The secretion of the classic inflammatory factors TNF-α and IL-6 was detected by ELISA. The HFD significantly upregulated the serum levels of TNF-α and IL-6 in mice. However, nicotine treatment significantly attenuated HFD-induced upregulation of serum TNF-α and IL-6 (Fig. 3).

Nicotine exerts anti-inflammatory effects via targeting a7 nAChR and inhibiting the NF-κB and ERK pathways

To investigate the underlying mechanism of the reduction of pro-inflammatory cytokines TNF-α and IL-6 by α7 nAChR activation, primary macrophages from the liver were isolated and assessed. Primary liver Kupffer cells were successfully isolated and identified by staining with CD11b and F480 macrophage-specific markers (Caltag Laboratories, Burlingame, CA, USA) and flow cytometric detection (BD-Accuri C6, BD Biosciences Franklin Lakes, NJ, USA) in comparison with murine macrophage RAW 264.7 cells (Fig. 4).

In the mouse model of NASH, the α7 nAChR agonist nicotine significantly attenuated HFD-induced upregulation of serum TNF-α and IL-6. To determine the anti-inflammatory mechanisms of α7 nAChR-activation, the present study assessed whether nicotine treatment blocked the production of TNF-α and IL-6 with or without α7 nAChR antagonist α-bungarotoxin (α-BGT) in the primary liver Kupffer cells. The secretion of the classic inflammatory factors TNF-α and IL-6 was detected by ELISA. Stimulation with lipopolysaccharide (LPS) significantly upregulated the secretion of TNF-α (Fig. 5A) and IL-6 (Fig. 5B) in the cell culture supernatants. However, nicotine treatment significantly attenuated LPS-induced upregulation of TNF-α (Fig. 5A) and IL-6 (Fig. 5B) in a dose-dependent manner. Furthermore, α7 nAChR antagonist α-BGT blocked the nicotine-induced reduction of TNF-α (Fig. 5A) and IL-6 (Fig. 5B), which indicated that nicotine reduced the production of TNF-α and IL-6 via activating α7 nAChR.

Release of inflammatory cytokines is mostly mediated via the ERK and NF-κB pathways, and ERK and NF-κB are the main downstream signaling molecules of α7 nAChR (27). The present study therefore investigated whether activation of α7 nAChR reduces the production of cytokines via inhibiting the ERK and NF-κB pathways (Fig. 5C). As shown in Fig. 5, nicotine treatment upregulated the protein levels of α7 nAChR in Kupffer cells in a dose-dependent manner, which was consistent with the results of a previous study (27). Furthermore, nicotine obviously downregulated ERK and NF-κB levels in Kupffer cells.

Discussion

The results present study suggested that specific interference with α7 nAChR represents a novel strategy for the treatment of NASH. It was shown that treatment with the α7 nAChR agonist nicotine for three weeks obviously attenuated hepatic steatosis and reduced the production of TNF-α and IL-6 in an HFD-induced mouse model of NASH. To investigate the underlying mechanism, the primary macrophages from mouse livers were isolated and treated with nicotine. The results showed that nicotine reduced LPS-induced secretion of TNF-α and IL-6 in vitro, which was blocked by α7 nAChR antagonist α-BGT. These results indicated that nicotine suppressed TNF-α and IL-6 secretion by LPS-stimulated macrophages through α7 nAChR activation. Furthermore, the present study showed that nicotine-stimulated α7 nAChR activation significantly downregulated NK-κB and ERK. It appeared that the activation of α7 nAChR suppressed the production of pro-inflammatory cytokines through NK-κB and ERK pathways.

NASH is increasingly recognized as a major epidemiological problem, linking the metabolic syndrome to liver fibrosis, cirrhosis and hepatocellular carcinoma. Currently discussed treatment options comprise drugs approved for managing the symptoms of impaired glucose metabolism, hypertension and hyperlipidemia, including angiotensin I antagonists or insulin sensitizers (30). However, the incidence of NAFLD in the human population is further increasing, affecting up to 30% of the general population worldwide, despite the availability of these drugs (31). In addition, the side effects of approved drugs preclude treatment of patient sub-populations, thus underlining the requirement for additional specific treatment options (27).

To test the effects of α7 nAChR activation on NASH, the HFD-induced mouse model of NASH was employed. The model developed symptoms within a time frame of 18 weeks and was characterized by a rather mild elevation in liver enzymes, such as ALT, in the circulation as well as the presence of lobular inflammation, which is also observed in humans with NASH (32). The present study showed that the α7 nAChR agonist nicotine reduced NASH-associated hepatic steatosis in mouse models. Furthermore, nicotine treatment decreased the secretion of the pro-inflammatory cytokines TNF-α, IL-6 in mice with NASH. This result indicated that activation of α7 nAChR and the resulting anti-inflammatory effects may represent a novel therapeutic strategy for NASH.

Inflammation characterized by the release of soluble factors, including chemokines and cytokines, in addition to immune cell activation, is regarded as an integral part of NASH and several lines of evidence suggested that targeting of inflammation is a promising tool for the management of NASH (29). The results of the present study showed that a7 nAChR agonist nicotine reduced the production of TNF-α and IL-6 in the mouse model of NASH in vivo and in primary Kupffer cells in vitro. These results were consistent with those of previous studies, which reported that activation of the a7 nAChR expressed on resident macrophages may suppress the local inflammation by reducing the production of pro-inflammatory cytokines TNF-α and IL-6 (27). Furthermore, the present study found that nicotine-induced a7 nAChR activation significantly inhibited the expression of NF-κB and ERK. This result indicated that activation of the a7 nAChR may inhibit cytokine production by Kupffer cells via the NF-κB and ERK pathways.

In conclusion, the present study indicated that modulating the inflammatory response in affected livers via activating a7 nAChR may represent a novel strategy for the treatment of NASH. The feasibility of this strategy requires pre-clinical and clinical validation in further studies.

Acknowledgments

This work was supported by the National Natural Science Foundation of China (grant no. 81100279) and the Foundation of Zhejiang Health Committee (no. 2013KYA145).

References

1 

Noureddin M, Yates KP, Vaughn IA, Neuschwander-Tetri BA, Sanyal AJ, McCullough A, Merriman R, Hameed B, Doo E, Kleiner DE, et al: Clinical and histological determinants of nonalcoholic steatohepatitis and advanced fibrosis in elderly patients. Hepatology. 58:1644–1654. 2013. View Article : Google Scholar : PubMed/NCBI

2 

Evans CD, Oien KA, MacSween RN and Mills PR: Non-alcoholic steatohepatitis: A common cause of progressive chronic liver injury? J Clin Pathol. 55:689–692. 2002. View Article : Google Scholar : PubMed/NCBI

3 

Ascha MS, Hanouneh IA, Lopez R, Tamimi TA, Feldstein AF and Zein NN: The incidence and risk factors of hepatocellular carcinoma in patients with nonalcoholic steatohepatitis. Hepatology. 51:1972–1978. 2010. View Article : Google Scholar : PubMed/NCBI

4 

Adams LA, Lymp JF, St Sauver J, Sanderson SO, Lindor KD, Feldstein A and Angulo P: The natural history of nonalcoholic fatty liver disease: A population-based cohort study. Gastroenterology. 129:113–121. 2005. View Article : Google Scholar : PubMed/NCBI

5 

Dunn W, Xu R, Wingard DL, Rogers C, Angulo P, Younossi ZM and Schwimmer JB: Suspected nonalcoholic fatty liver disease and mortality risk in a population-based cohort study. AM J Gastroenterol. 103:2263–2271. 2008. View Article : Google Scholar : PubMed/NCBI

6 

Wong VW, Wong GL, Tsang SW, Fan T, Chu WC, Woo J, Chan AW, Choi PC, Chim AM, Lau JY, et al: High prevalence of colorectal neoplasm in patients with non-alcoholic steatohepatitis. Gut. 60:829–836. 2011. View Article : Google Scholar : PubMed/NCBI

7 

Gastaldelli A, Kozakova M, Højlund K, Flyvbjerg A, Favuzzi A, Mitrakou A and Balkau B: Fatty liver is associated with insulin resistance, risk of coronary heart disease and early atherosclerosis in a large European population. Hepatology. 49:1537–1544. 2009. View Article : Google Scholar : PubMed/NCBI

8 

Bilici A, Ozguroglu M, Mihmanli I, Turna H and Adaletli I: A case-control study of non-alcoholic fatty liver disease in breast cancer. Med Oncol. 24:367–371. 2007. View Article : Google Scholar : PubMed/NCBI

9 

Pagadala MR, Zein CO, Dasarathy S, Yerian LM, Lopez R and McCullough AJ: Prevalence of hypothyroidism in nonalcoholic fatty liver disease. Dig Dis Sci. 57:528–534. 2012. View Article : Google Scholar

10 

Caballería L, Auladell MA, Torán P, Miranda D, Aznar J, Pera G, Gil D, Muñoz L, Planas J, Canut S, et al: Prevalence and factors associated with the presence of non alcoholic fatty liver disease in an apparently healthy adult population in primary care units. BMC Gastroenterol. 7:412007. View Article : Google Scholar : PubMed/NCBI

11 

Gelpi Méndez JA, Castellanos Fillot A, Sainz Gutiérrez JC, Quevedo Aguado L and Martin Barallat J: Prevalence of non-alcoholic fatty liver disease and associated risk factors among managers from the community of Madrid. Arch Prev Riesgos Labor. 17:84–90. 2014.In Spanish. View Article : Google Scholar

12 

Leite NC, Salles GF, Araujo AL, Villela-Nogueira CA and Cardoso CR: Prevalence and associated factors of non-alcoholic fatty liver disease in patients with type-2 diabetes mellitus. Liver Int. 29:113–119. 2009. View Article : Google Scholar

13 

Loria P, Lonardo A, Lombardini S, Carulli L, Verrone A, Ganazzi D, Rudilosso A, D'Amico R, Bertolotti M and Carulli N: Gallstone disease in non-alcoholic fatty liver: Prevalence and associated factors. J Gastroen Hepatol. 20:1176–1184. 2005. View Article : Google Scholar

14 

Radu C, Grigorescu M, Crisan D, Lupsor M, Constantin D and Dina L: Prevalence and associated risk factors of non-alcoholic fatty liver disease in hospitalized patients. J Gastrointestin Liver Dis. 17:255–260. 2008.PubMed/NCBI

15 

Hu KC, Wang HY, Liu SC, Liu CC, Hung CL, Bair MJ, Liu CJ, Wu MS and Shih SC: Nonalcoholic fatty liver disease: Updates in noninvasive diagnosis and correlation with cardiovascular disease. World J Gastroenterol. 20:7718–7729. 2014. View Article : Google Scholar : PubMed/NCBI

16 

Sanyal AJ: NASH: A global health problem. Hepatol Res. 41:670–674. 2011. View Article : Google Scholar : PubMed/NCBI

17 

Takaki A, Kawai D and Yamamoto K: Molecular mechanisms and new treatment strategies for non-alcoholic steatohepatitis (NASH). Int J Mol Sci. 15:7352–7379. 2014. View Article : Google Scholar : PubMed/NCBI

18 

Musso G, Anty R and Petta S: Antioxidant therapy and drugs interfering with lipid metabolism: could they be effective in NAFLD patients? Curr Pharm Des. 19:5297–5313. 2013. View Article : Google Scholar : PubMed/NCBI

19 

Yalcin M, Akarsu M, Celik A, Sagol O, Tunali S, Ertener O, Bengi G and Akpinar H: A comparison of the effects of infliximab, adalimumab, and pentoxifylline on rats with non-alcoholic steatohepatitis. Turk J Gastroenterol. 25:167–175. 2014. View Article : Google Scholar

20 

Fock KM and Khoo J: Diet and exercise in management of obesity and overweight. J Gastroenterol Hepatol. 28:59–63. 2013. View Article : Google Scholar : PubMed/NCBI

21 

Beaton MD: Current treatment options for nonalcoholic fatty liver disease and nonalcoholic steatohepatitis. Can J Gastroenterol. 26:353–357. 2012.PubMed/NCBI

22 

Malaguarnera M, Di Rosa M, Nicoletti F and Malaguarnera L: Molecular mechanisms involved in NAFLD progression. J Mol Med (Berl). 87:679–695. 2009. View Article : Google Scholar

23 

Satapathy SK, Garg S, Chauhan R, Sakhuja P, Malhotra V, Sharma BC and Sarin SK: Beneficial effects of tumor necrosis factor-alpha inhibition by pentoxifylline on clinical, biochemical and metabolic parameters of patients with nonalcoholic steatohepatitis. AM J Gastroenterol. 99:1946–1952. 2004. View Article : Google Scholar : PubMed/NCBI

24 

Mitchel EB and Lavine JE: Review article: the management of paediatric nonalcoholic fatty liver disease. Aliment Pharmacol Ther. 40:1155–1170. 2014. View Article : Google Scholar : PubMed/NCBI

25 

Filippini P, Cesario A, Fini M, Locatelli F and Rutella S: The Yin and Yang of non-neuronal α7 -nicotinic receptors in inflammation and autoimmunity. Curr drug targets. 13:644–655. 2012. View Article : Google Scholar : PubMed/NCBI

26 

Boeckxstaens G: The clinical importance of the anti-inflammatory vagovagal reflex. Handb Clin Neurol. 117:119–134. 2013. View Article : Google Scholar : PubMed/NCBI

27 

Ganz M and Szabo G: Immune and inflammatory pathways in NASH. Hepatol int. 7:771–781. 2013. View Article : Google Scholar

28 

Meli R, Mattace Raso G and Calignano A: Role of innate immune response in non-alcoholic Fatty liver disease: metabolic complications and therapeutic tools. Front Immunol. 5:1772014. View Article : Google Scholar : PubMed/NCBI

29 

Braunersreuther V, Viviani GL, Mach F and Montecucco F: Role of cytokines and chemokines in non-alcoholic fatty liver disease. World J Gastroenterol. 18:727–735. 2012. View Article : Google Scholar : PubMed/NCBI

30 

Weiß J, Rau M and Geier A: Non-alcoholic fatty liver disease epidemiology, clinical course, investigation, and treatment. Dtsch Arztebl Int. 111:447–452. 2014.

31 

Ratziu V, Goodman Z and Sanyal A: Current efforts and trends in the treatment of NASH. J Hepatol. 62:S65–S75. 2015. View Article : Google Scholar : PubMed/NCBI

32 

Gadd VL, Skoien R, Powell EE, Fagan KJ, Winterford C, Horsfall L, Irvine K and Clouston AD: The portal inflammatory infiltrate and ductular reaction in human nonalcoholic fatty liver. Hepatology. 59:1393–1405. 2014. View Article : Google Scholar

Related Articles

Journal Cover

November-2015
Volume 12 Issue 5

Print ISSN: 1791-2997
Online ISSN:1791-3004

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
x
Spandidos Publications style
Zhou Z, Liu YC, Chen XM, Li FQ, Tong XJ, Ding YP and Tang CL: Treatment of experimental non‑alcoholic steatohepatitis by targeting α7 nicotinic acetylcholine receptor‑mediated inflammatory responses in mice. Mol Med Rep 12: 6925-6931, 2015.
APA
Zhou, Z., Liu, Y., Chen, X., Li, F., Tong, X., Ding, Y., & Tang, C. (2015). Treatment of experimental non‑alcoholic steatohepatitis by targeting α7 nicotinic acetylcholine receptor‑mediated inflammatory responses in mice. Molecular Medicine Reports, 12, 6925-6931. https://doi.org/10.3892/mmr.2015.4318
MLA
Zhou, Z., Liu, Y., Chen, X., Li, F., Tong, X., Ding, Y., Tang, C."Treatment of experimental non‑alcoholic steatohepatitis by targeting α7 nicotinic acetylcholine receptor‑mediated inflammatory responses in mice". Molecular Medicine Reports 12.5 (2015): 6925-6931.
Chicago
Zhou, Z., Liu, Y., Chen, X., Li, F., Tong, X., Ding, Y., Tang, C."Treatment of experimental non‑alcoholic steatohepatitis by targeting α7 nicotinic acetylcholine receptor‑mediated inflammatory responses in mice". Molecular Medicine Reports 12, no. 5 (2015): 6925-6931. https://doi.org/10.3892/mmr.2015.4318