ABSTRACT
@#To explore the role of autophagy in hair follicle cycle and whether 3-methyladenine(3-MA)could promote hair regeneration in C57BL/6 mice through inhibiting autophagy flux, hair regeneration model of C57BL/6 mice was induced on the dorsal skin by depilation, and 3-MA was intraperitoneally injected to investigate hair regrowth, meanwhile vehicle and rapamycin(RAPA)were used as the controls. Results showed that 3-MA could obviously promote hair regrowth in depilated C57BL/6 mice. Furtherly, haematoxylin-eosin staining, immunohistochemical staining, immunofluorescence and Western blot tests were used to investigate the autophagy signals and the cell proliferation. Results showed that the expression of Beclin1 and LC3B II/I ratio were significantly decreased. Expression of P62 and Ki67 were increased, as well as the CD34 and CD49f double-labeled hair follicle stem cells were obviously increased inside bulge areas in 3-MA group, while contrarily in RAPA group. These results affirmed 3-MA, an autophagy inhibitor, could promote hair regeneration in depilated C57BL/6 mice by facilitating the transformation of hair follicle from telogen to anagen. 3-MA and other analogous autophagy inhibitors probably have a potential usage in future therapy in human telogen effluvium diseases.
ABSTRACT
This study was designed to examine the effects of autophagy on the lipopolysaccharide(LPS)-induced acute lung injury (ALI)and to analyze the possible mechanism.Kunming mice of clean grade were randomly divided into 6 groups:control group(saline;ip);model group(LPS 10 mg/kg;ip);RAP group(rapamycin 6 mg/kg;ip);RAP+LPS group (RAP 6 mg/kg and LPS 10 mg/kg;ip);3-MA group (3-methyladenine 300 μg/kg;ip);and 3-MA+LPS group (3-MA 300 μg/kg and LPS 10 mg/kg;ip).6 h after LPS injection;mortality was checked.Neutrophil aggregation;and the expressions of TNF-α;LC3 and P62 in lung tissue were checked by fluo-rescence microscopy and Western blot.The results revealed a higher mortality;neutrophil infiltration and TNF-αexpression and significantly increased levels of autophagy marker LC3-II and P62 in LPS group;in RAP+LPS group;pretreatment with RAP notably reversed LPS-induced neutrophil infiltration and TNF-αexpression;LC3-II were further slightly increased;while P62 was significantly decreased;in 3-MA+LPS group;pretreatment of 3-MA slightly decreased LC3-II expression;P62;neutrophil infiltration and TNF-αremained almost the same to those in LPS group.These data suggesed that;in sepsis;autophagy flux in the lung tissue is partially blocked;and RAP help to alleviate LPS-induced lung injury;which might through promoting autophagosome maturation;smoothing autophagy flux;and eventually to mitigate pulmonary inflammation.
ABSTRACT
Objective To observe the expressions of two inhibitors of apoptosis proteins (LAPs), survivin and livin, as well as their related factors, Bci-xl and Caspase-3 in mycosis fungoides (MF), along with the effects of NB-UVB irradiation on them. Methods Totally, 30 patients with MF (5 at erythema stage, 16 at plaque stage and 9 at tumor stage) collected from 1995 to 2007 were included into this study. Of the patients, 11 received the treatment with NB-UVB irradiation. Tissue samples were resected from 30 untreated patients, 11 irradiated patients and 10 normal human controls. SABC immunohistochemistry (IHC) stain was used to evaluate the protein expression of survivin, livin, Bel-xi and caspase-3 in these samples. Also, the mRNA expression of these four factors and cell apoptosis were detected by hybridization in situ (ISH) and terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-fluorescence nick end labeling (TUNEL stain) respectively in the samples from 11 patients before and after NB-UVB irradiation. Results In samples of erythema-stage MF, plaque-stage MF and tumor-stage MF, the positivity rate was 40.00%, 75.00%, 77.78% for survivin respectively, 60.00%, 68.75%, 88.89% for Bcl-xl respectively, 40%, 25%, 44.44% for livin respectively, and 60.00%, 68.75%, 88.89% for caspase-3 respectively. No expression of survivin or Bcl-xl was observed in normal controls, while the expression of livin and caspase-3 was similar between MF and control samples. After NB-UVB irradiation, an increase was noticed in the count of apoptosis cells (t=6.49, P<0.001) and mRNA expression of caspase-3 (P<0.10), while a decrease in the mRNA expression of survivin and Bcl-xl in MF tissues, and no changes occurred to the mRNA expression of livin (P>0.10). Conehsions Survivin, Bcl-xl and caspase-3 may be associated with the pathogenesis of MF by regulating the cell apoptosis of T lymphocytes. NB-UVB could suppress the mRNA expression of survivin and Bcl-xl, lower the levels of LAP, enhance the transcription of caspase-3, and accelerate the apoptosis of tumor cells, which may partly explain the mechanism of therapeutic effect of NB-UVB in MF.