Phillygenin from Forsythia suspensa leaves exhibits analgesic potential and anti-inflammatory activity in carrageenan-induced paw edema in mice.

Forsythia suspensa (Thunb.) Vahl (Oleaceae) leaves are valuable sources of phillygenin. This study aimed to isolate phillygenin from F. suspensa leaves and examine its analgesic and anti-inflammatory effects. Phillygenin was successfully extracted and isolated from F. suspensa leaves after fermentation. Phillygenin significantly reduced the number of writhing induced by acetic acid, prolonged the latency period in the hot plate test, and inhibited the xylene-induced ear edema and carrageenan-induced paw edema in mice. IL-6, TNF-α, IL-1ß, NO, and PGE2 levels in the carrageenan-induced paw edema were notably reduced after pretreatment with phillygenin. Phillygenin significantly decreased the iNOS and COX-2 protein expressions and the IκB-α and NF-κB p65 phosphorylation. This study demonstrated that phillygenin is a potential therapeutic candidate for managing pain and inflammation-mediated disorders. The study contributes to the comprehensive development and utilization of F. suspensa leaves for economic and health care. PRACTICAL APPLICATIONS: Phillygenin is one of the major active ingredients in Forsythia suspensa. But the content of phillygenin in F. suspensa is very low which limits its application. Phillygenin has potential pharmacological activity and anti-inflammatory properties. However, the potential effects of phillygenin on analgesic activity have not been clarified. Furthermore, the data on its anti-inflammatory activity in vivo are relatively limited. This study evaluated the analgesic activity for the first time and the acute anti-inflammatory effect of phillygenin from F. suspensa leaves by fermentation, which indicated phillygenin is a potential therapeutic candidate for managing pain and inflammation-mediated disorders.

Altered microRNA-9 Expression Level is Directly Correlated with Pathogenesis of Nonalcoholic Fatty Liver Disease by Targeting Onecut2 and SIRT1.

BACKGROUND MicroRNA-9 (miR-9) was detected in nonalcoholic fatty liver disease (NAFLD) patients to understand the role of miR-9 in NAFLD development. MATERIAL AND METHODS Between February 2014 and February 2015, 105 cases of NAFLD were recruited and confirmed by liver biopsy pathology, including patients with mild NAFLD (n=58) and moderate-severe NAFLD (n=47); nonalcoholic steatohepatitis (NASH) (n=53) and non-NASH (n=52); and 50 healthy participants were regarded as the healthy control group. MiR-9 expression was measured by qRT-PCR. For in vitro experiments, L-02 normal liver cells were divided into normal control group (cultured with original culture medium), dimethyl sulfoxide (DMSO) group (cultured with DMSO) and oleic acid group (cultured with oleic acid to induce fatty change), and MTT assay was used to measure the effect of different oleic acid concentrations on cell proliferation. Nile red staining was used to detect intracellular accumulation of lipid droplets. Further, synthetic miR-9 mimic and its control and miR-9 inhibitors and its control were independently transfected into L-02 cells. RESULTS MiR-9 levels in the mild NAFLD group and moderate-severe NAFLD group were significantly higher than in the healthy control group (both P<0.05). Mean fluorescence intensity of lipid droplets increased with the duration of induction, and were dramatically higher in oleate-treated L-02 cells; intracellular triglyceride (TG) content was also higher. miR-9 levels significantly increased following oleate induction. Importantly, miR-9 levels were significantly elevated upon miR-9 mimic transfection. Conversely, miR-9 level was lowered with miR-9 inhibitors transfection. Additionally, Onecut2 and SIRT1 were identified as miR-9 targets. CONCLUSIONS A positive relationship between miR-9 and steatosis was established with our results that miR-9 mimic transfection decreased intracellular lipid content. Finally, we identified 2 miR-9 targets, Onecut2 and SIRT1, which may be crucial players in NAFLD development.

AdipoQ T45 G and G276 T Polymorphisms and Susceptibility to Nonalcoholic Fatty Liver Disease Among Asian Populations: A Meta-Analysis and Meta-Regression.

BACKGROUND: Adiponectin (AdipoQ) is an adipose-derived plasma protein that plays an important role in hepatic lipoprotein-lipid metabolism. Emerging evidence have shown that two common polymorphisms (T45 G and G276 T) in the AdipoQ gene may contribute to increasing susceptibility to nonalcoholic fatty liver disease (NAFLD); however individually published studies show inconclusive results. This meta-analysis aimed to derive a more precise estimation of the association of AdipoQ T45 G (rs2241766 T>G) and G276 T (rs1501299 G>T) polymorphisms with NAFLD risk. METHOD: Potential relevant studies were identified covering the following databases: PubMed, Embase, Web of Science, the Cochrane Library, Chinese National Knowledge Infrastructure (CNKI), Chinese Bio-medicine Database (CBM), and Chinese Sci-tech Journals databases. Statistical analyses were calculated using the version 12.0 STATA software (Stata Corp, College Station, TX, USA). Odds ratios (ORs) and its corresponding 95% confidence interval (CI) were calculated. RESULT: Ten case-control studies were included with a total of 2,672 subjects, of these 1,117 being NAFLD patients and 1,555 being healthy controls. Our meta-analysis results revealed that the T variant of AdipoQ rs2241766 T>G polymorphism may be associated with an increased risk of NAFLD. There was also a significant association between the G variant of AdipoQ rs1501299 G>T polymorphism and an increased risk of NAFLD. Country-stratified analysis indicated that a higher AdipoQ rs2241766 T>G polymorphism was closely related with an increased risk of NAFLD in Chinese and Indian populations (all Ps < 0.05); a similar result was observed in Chinese populations between AdipoQ rs2241766 T>G polymorphism and an increased risk of NAFLD (P < 0.05). CONCLUSION: In conclusion, the current meta-analysis indicates that AdipoQ rs2241766 T>G and rs1501299 G>T polymorphisms may contribute to an increasing susceptibility to NAFLD. Moreover, this meta-analysis also suggests for future larger studies with stratified case-control population, and greater focus on the gene-environment interactions regarding NAFLD susceptibility for valid studies.

USF-1 genetic polymorphisms confer a high risk of nonalcoholic fatty liver disease in Chinese population.

Genetic polymorphisms in upstream transcription factor 1 (USF1) were investigated for their links to increased risk of nonalcoholic fatty liver disease (NAFLD) in Chinese population. Between January 2013 and April 2014, 174 patients with NAFLD in the First Affiliated Hospital of China Medical University were selected for this study. A group of 100 healthy subjects were identified as the control group. The MALDI-TOF-MS, a mass spectrometry based technique, was used to detect USF-1 genetic polymorphisms using PCR amplified DNA products. Furthermore, Automatic Chemistry Analyzer (ACA) was used to determine the clinical indicators. Genotypes, allele frequencies and clinical indicators were measured to assess NAFLD risk in relation to the SNPs. USF-1 rs6427573 genetic polymorphisms were associated with an increased risk of NAFLD (AA vs. GG: OR = 3.16, 95% CI = 1.56-6.43, P = 0.001; GA + AA vs. GG: OR = 1.87, 95% CI = 1.13-3.09, P = 0.015; GG + AA vs. AA: OR = 2.96, 95% CI = 1.49-5.88, P = 0.001; G vs. A: OR = 2.10, 95% CI = 1.43-3.09, P < 0.001). Similarly, rs2516839 polymorphisms also conferred a risk for NAFLD (AA vs. GG: OR = 2.49, 95% CI = 1.43-4.34, P = 0.001; GA + AA vs. GG: OR = 1.69, 95% CI = 1.02-2.78, P = 0.041). On the other hand, rs3737787 and rs2774279 showed no statistical significances in the NAFLD group and control group (P > 0.05). Two USF-1 genetic polymorphisms, rs6427573 and rs2516839, may present an increased risk of NAFLD.

Effects of ethanol on the expression of caveolin-1 in HepG2 cells.

This study aimed to investigate the effects of ethanol on the expression of caveolin­1 (CAV­1) in HepG2 hepatocarcinoma cells. Ethanol­treated HepG2 cells were investigated using the in vitro model to determine whether ethanol can influence the expression of CAV­1. Cell viability was measured using the colorimetric 3­(4, 5­dimethylthiazol­2­yl)­2,5­diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays. Expression of CAV­1 was detected using western blot analysis. Quantitative PCR (qPCR) was used to determine CAV­1 mRNA levels. The distribution of CAV­1 in HepG2 cells was analyzed using immunofluorescence. The MTT assay results revealed that cell viability was not altered at ethanol concentrations of <1.0%, while ethanol concentrations >1.0% caused cell shedding, but not cell fragmentation. Western blot analysis showed significant differences in the levels of CAV­1 expression between the control group and the 1.0% ethanol­treated group at 6, 12 and 24 h (all P<0.05). qPCR showed significant differences in the expression levels of caveolin­1 mRNA between the control group and the 1.0% ethanol­treated group at 6 h, 12 h and 24 h (all P<0.05). Immunofluorescence demonstrated that CAV­1 was distributed discontinuously at the boundaries of HepG2 cells. The results indicate that ethanol may increase the expression of CAV­1 in HepG2 cells.

Relationship of SREBP-2 rs2228314 G>C polymorphism with nonalcoholic fatty liver disease in a Han Chinese population.

OBJECTIVE: This study aims at investigating the relationship of SREBP-2 rs2228314 G>C polymorphism with the risk of nonalcoholic fatty liver disease (NAFLD) in a Han Chinese population. METHOD: This case-control study was conducted at the First Affiliated Hospital of China Medical University. Three-hundred subjects who met the diagnostic criteria of NAFLD and had typical clinical and ultrasonographic findings were placed in the case group. There were 160 matched healthy controls in the control group. A common single nucleotide polymorphism (SNP) (rs2228314 G>C) in the SREBP-2 gene was tested. Genetic analyses were performed using genomic DNA extracted from peripheral blood leukocytes. Polymerase chain reaction-restriction fragment length polymorphism was applied to detect SNP. RESULTS: Our results indicated that the GG genotype and G carrier (CG+GG) of rs2228314 G>C polymorphism in the SREBP-2 gene were strongly associated with susceptibility to NAFLD (both p<0.001). However, there was no significant difference in the frequency of G allele between NAFLD patients and healthy controls (p=0.328). Multivariate logistic regression analysis revealed that GG genotype, G carrier, body mass index, high-density lipoprotein cholesterol, total cholesterol, alanine aminotransferase, and γ-glutamyl-transferase might be associated with an increased risk of NAFLD (all p<0.05). CONCLUSION: The results of this study provide evidence that the GG genotype and G carrier (CG+GG) of rs2228314 G>C polymorphism in the SREBP-2 gene may increase the risk of NAFLD.

Genetic polymorphisms in the IL-18 gene and ulcerative colitis risk: a meta-analysis.

This meta-analysis was performed to evaluate the relationships between genetic polymorphisms in the IL-18 gene and ulcerative colitis (UC) risk. The PubMed, CISCOM, CINAHL, Web of Science, Google Scholar, EBSCO, Cochrane Library, and CBM databases were searched for relevant articles published before November 1st, 2013 without any language restrictions. Meta-analysis was conducted using the STATA 12.0 software. Crude odds ratios (ORs) with their 95% confidence intervals (95% CI) were calculated. Eight case-control studies with a total of 1000 UC cases and 1392 healthy subjects met the inclusion criteria. Six common polymorphisms in the IL-18 gene were evaluated, including rs1946518 A>C, rs187238 G>C, rs917997 G>A, Codon35, rs1946519 C>A, and rs360718 A>C. The results of our meta-analysis suggest that the IL-18 rs1946518 (allele model: OR=1.22, 95% CI: 1.01-1.48, p=0.039; dominant model: OR=1.44, 95% CI: 1.01-2.06, p=0.045; respectively), rs187238 (allele model: OR=1.38, 95% CI: 1.19-1.61, p<0.001; dominant model: OR=1.50, 95% CI: 1.03-2.19, p=0.034; respectively), and rs360718 (allele model: OR=2.18, 95% CI: 1.22-3.90, p=0.008) polymorphisms might be strongly correlated with an increased risk of UC. A subgroup analysis was conducted to investigate the effect of ethnicity on an individual's risk of UC. Our results revealed positive significant correlations between IL-18 genetic polymorphisms and an increased risk of UC among Asians (allele model: OR=1.36, 95% CI: 1.16-1.60, p<0.001; dominant model: OR=1.50, 95% CI: 1.14-1.98, p=0.004; respectively) and Africans (allele model: OR=1.45, 95% CI: 1.03-2.05, p=0.034), but not among Caucasians (all p>0.05). Our findings provide convincing evidence that IL-18 genetic polymorphisms may contribute to susceptibility to UC, especially the rs1946518, rs187238, and rs360718 polymorphisms among Asians and Africans.