Curcumin Promotes Apoptosis of Activated Hepatic Stellate Cells by Inhibiting Protein Expression of the MyD88 Pathway.

Activation and proliferation of hepatic stellate cells (HSC) play an important role in the progress of liver fibrosis. HSC activation occurs in response to inflammatory cytokines, cellular interactions with immune cells, and morphogenetic signals. The literature hints to a role of the adaptor protein MyD88 in fibrosis. Although curcumin has been shown to exert inhibitory effects on the proliferation of HSC in vitro, its influence on the MyD88 pathway in HSC has remained unclear. Here, we investigated whether curcumin accelerates apoptosis of HSC through the MyD88 pathway. HSC (rat HSC T6) were divided into a control group, MyD88 small interfering RNA (siRNA) group, curcumin group, and curcumin + MyD88 siRNA group. The MyD88 siRNA groups were exposed to siRNA for 48 h. The curcumin groups were cultured in the presence of curcumin for 24 h. Apoptosis was detected by flow cytometry. For Toll-like receptor (TLR) 2 and 4 as well as MyD88 and the dependent factors NF-κB, TNF-α, and IL-1ß, mRNA expression was detected by reverse transcription polymerase chain reaction (RT-PCR). For MyD88, protein expression was further observed by Western Blot. Both curcumin and MyD88 siRNA inhibited the mRNA expression of MyD88 pathway-related effectors (TLR2, TLR4, NF-κB, TNF-α, IL-1ß) in HSC. Furthermore, both treatments reduced the expression of MyD88 protein in HSC and promoted their apoptosis. These effects were more obvious in the curcumin + MyD88 siRNA group. This study demonstrates that curcumin promotes apoptosis of activated HSC by inhibiting the expression of cytokines related to the MyD88 pathway. It elucidates the possible mechanisms of curcumin in inducing apoptosis of HSC through the MyD88 pathway.

Effect of acupuncture treatment on vascular cognitive impairment without dementia: study protocol for a randomized controlled trial.

BACKGROUND: Vascular cognitive impairment, no dementia (VCIND) is a condition at risk for future dementia and should be the target of preventive strategies. Preliminary evidence suggests that acupuncture may be a clinically effective intervention for people with early-stage vascular cognitive impairment. We will do a multicenter, 6-month, drug-controlled, nonblinded, randomized, parallel-group trial to determine whether acupuncture is effective for improving cognitive function and quality of life for patients with VCIND. METHODS/DESIGN: A total of 216 eligible patients will be recruited and randomly assigned acupuncture for two sessions/week (n = 108) or citicoline 300 mg/day (n = 108) in a multicenter, 6-month trial. The primary endpoint is cognition (Alzheimer's Disease Assessment Scale, Cognitive Subscale (ADAS-cog)). Secondary endpoints include assessments of activities of daily living and behavioral symptoms (Clock Drawing Test (CDT), Activities of Daily Living (ADL) and Instrumental Activities of Daily Living scale (IADL)). DISCUSSION: This will be the first large-scale trial specifically evaluating acupuncture therapy in VCIND. If the study confirms the effectiveness and safety of acupuncture treatment, it will be important to examine how the acupuncture approach could most effectively be integrated into the provision of routine healthcare. TRIAL REGISTRATION: This study is registered as an International Standard Randomised Controlled Trial on 17 January 2014, number ISRCTN 82980206.

Transforming growth factor­ß1 reduces apoptosis via autophagy activation in hepatic stellate cells.

Autophagy is a metabolic process that is important in fibrogenesis, in which cellular components are degraded by lysosomal machinery. Transforming growth factor ß1 (TGF­ß1) is a potent fibrogenic cytokine involved in liver fibrosis; however, it remains elusive whether autophagy is regulated by TGF­ß1 in this process. In the present study, the function of TGF­ß1­mediated autophagy in the proliferation and apoptosis of hepatic stellate cells (HSCs) was investigated. A rat HSC cell line (HSC­T6) was incubated with or without TGF­ß1 followed by bafilomycin A1, and microtubule-associated proteins 1A/1B light chain 3 (LC3) small interfering (si)RNA was used to inhibit autophagy in order to assess the association between TGF­ß1 and autophagy. HSC­T6 cell transient transfection was accomplished with a pLVX­AcGFP­N1­rLC3B­encoding plasmid. An MTS assay and flow cytometry were utilized to detect proliferation and apoptosis of HSC­T6 cells. Quantitative polymerase chain reaction, immunofluorescence and western blot analysis were used to detect the presence of activation markers. Proliferation was increased and apoptosis was reduced in HSC­T6 cells treated with TGF­ß1 compared with cells subjected to serum deprivation. However, when HSC­T6 cells were treated with bafilomycin A1 and LC3 siRNA, increased apoptosis and reduced proliferation were observed. In addition, protein and mRNA expression levels of the autophagy marker LC3 were significantly increased. GFP­LC3 punctate markings were more prolific following TGF­ß1 treatment of HSC­T6 cells, indicating that TGF­ß1 may rescue HSC­T6 cells from serum deprivation and reduce apoptosis via autophagy induction. The present study elucidated the possible functions of TGF­ß1­mediated autophagy in the pathological process of liver fibrosis.

ß-Nerve growth factor attenuates hepatocyte injury induced by D-galactosamine in vitro via TrkA NGFR.

Nerve growth factor (NGF) regulates the proliferation, differentiation and survival of cells and is also involved in the wound healing and tissue remodeling processes. The biological effects of NGF are dependent upon receptor signal-mediating functions, which differ between cells. This study attempted to investigate the hepatoprotective effect and possible mechanism of ß-NGF on D-galactosamine (D-GalN)-injured human liver L-02 cell lines. We demonstrated that L-02 cells expressed the neurotrophin receptors tyrosine kinase-A nerve growth factor receptor (TrkA NGFR) and p75 pan-neurotrophin receptor (p75NTR). Recombinant human ß-NGF markedly reduced cell injury and promoted the proliferation of L-02 cells damaged by D-GalN. However, this proliferation effect was blocked by the anti-TrkA NGFR antibody. Lactate dehydrogenase (LDH) and malondialdehyde (MDA) were released at reduced levels in the L-02 cell culture supernatant pretreated with ß-NGF. Furthermore, the albumin (ALB) content in the cell medium and intracellular glutathione (GSH) levels were markedly augmented, and the permeability of the mitochondrial membrane of the L-02 cells was improved by ß-NGF. Our results suggested that exogenous ß-NGF protects L-02 cells from D-GalN-induced injury through the NGF/TrkA NGFR signaling pathway.

[Expression of eotaxin mRNA and TNF-alpha mRNA in lung tissues of sensitized mice and modulation by anti-inflammatory drugs].

OBJECTIVE: To establish determination methods of eotaxin mRNA and TNF-alpha mRNA expression in the lung tissue of mice. METHODS: Eotaxin mRNA and TNF-alpha mRNA expressions were determined by semi-quantitative RT-PCR. The functional implications of eotaxin mRNA and TNF-alpha mRNA expression were examined by detecting the numbers of total leucocytes and eosinophils in bronchoalveolar lavage fluid(BALF). RESULT: Eotaxin mRNA and TNF-alpha mRNA expression in lung tissue total numbers of leucocyte and numbers of eosinophil in BALF increased in sensitized mice compared with those in normal mice. Dexamethasone significantly but did not inhibit eotaxin mRNA and TNF-alpha mRNA expressions, and eosinophil infiltration in the lungs of the sensitized mice. A compound preparation of traditional Chinese medicine inhibited eotaxin mRNA and eosinophil infiltration, influenced TNF-alpha mRNA expression. CONCLUSION: Increased eotaxin mRNA expression in lung tissue is associated with eosinophil infiltration in BALF, which indicates that the methods of semi-quantitative RT-PCR may be useful to the study of the mechanism of antiasthmatic medicine.