ABSTRACT
ObjectiveThe lung mesenchymal stem cells （LMSCs） induced by D-galactose （D-gal） were intervened by Wenfei Huaxian decoction-containing serum to explore the mechanism of Wenfei Huaxian decoction in delaying the senescence of LMSCs through the nicotinamide phosphoribosyltransferase/silent information regulator 1 （NAMPT/SIRT1） signaling pathway. MethodWenfei Huaxian decoction-containing serum was prepared. LMSCs were isolated by gradient density centrifugation， and they were cultured and identified in vitro. The senescence model in vitro was established by stimulating cells via D-gal for 24 h. LMSCs cells were modeled after being treated with different volume fractions （5%， 10%， 20%， 40%， and 80%） of Wenfei Huaxian decoction-containing serum for 24 h， and the cell proliferation level was detected by methyl thiazolyl tetrazolium （MTT） method. The cells were randomly divided into blank serum group， model group， and high， medium， and low dose groups of Wenfei Huaxian decoction-containing serum. Senescence-associated β-galactosidase （SA-β-gal） staining was used to detect the senescence of LMSCs in each group. The content of NAD + was detected by colorimetry. The levels of senescence-associated factors （p16 and p53）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α） in cell culture supernatant were detected by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the relative expression of senescence-associated proteins and NAMPT/SIRT1 signaling pathway-related proteins. ResultCompared with the blank serum group， the proliferation of LMSCs was significantly inhibited after D-gal stimulation for 24 h （P<0.01）. Compared with the model group， the proliferation of LMSCs could be promoted after intervention with the corresponding Wenfei Huaxian decoction-containing serum （P<0.05， P<0.01）. Compared with the blank serum group， the SA-β-gal staining of LMSCs in the model group after D-gal stimulation was enhanced， and the content of NAD+ was increased （P<0.01）. The expression levels of senescence factors p16 and p53， as well as SASP pro-inflammatory factors IL-6 and TNF-α in the cell culture supernatant， were significantly increased （P<0.01）. The expression of senescence-associated proteins p16， p21， and p53 increased （P<0.01）， and the protein expression of NAMPT， SIRT1， peroxisome proliferator-activated receptor γ coactivator 1α （PGC-1α）， and forkhead box family transcription factor O1 （FoxO1） decreased （P<0.01）. Compared with the model group， the SA-β-gal staining of LMSCs in each group of Wenfei Huaxian decoction-containing serum was significantly reduced， and the content of NAD+ was decreased （P<0.01）. The senescence factors （p16 and p53） and inflammatory factors （IL-6 and TNF-α） in the cell culture supernatant were significantly decreased （P<0.01）. The expression of senescence-associated proteins （P16， P21， and P53） decreased （P<0.05， P<0.01）. The protein expressions of NAMPT， SIRT1， PGC-1α， and FoxO1 were significantly up-regulated （P<0.05， P<0.01）. ConclusionWenfei Huaxian decoction can alleviate senescence and inflammatory response damage of D-gal-induced LMSCs， and its mechanism may be related to the regulation of the NAMPT/SIRT1 signaling pathway.
ABSTRACT
This study was aimed to explore the molecular mechanism of nano-realgar in treatment of systematic lupus erythematosus (SLE) lupus nephritis (LN). Intragastric administration of equal volume of high-, middle-, low-dose nano-realgar suspension, and normal saline (NS) were given to MRL/lpr mice, respectively. The observations were made on levels of ANA, ds-DNA antibody, IgG and IgM in blood serum, as well as TWEAK, NF-κB, MCP-1 mRNA and protein expression levels in renal tissues. The results showed that compared with the NS group, levels of ANA, ds-DNA antibody, IgG and IgM were obviously reduced (P < 0.05); the levels of TWEAK, NF-κB and MCP-1 mRNA were obviously reduced (P < 0.05); the protein expression levels of TWEAK, NF-κB and MCP-1 mRNA were obviously reduced (P < 0.05) in the high-, middle-, low-dose nano-realgar group. It was concluded that nano-realgar intervened the TWEAK-NF-κB signal pathway through downregulating MCP-1 expression among MRL/lpr mice, in order to reduce the levels of ANA, ds-DNA antibody, IgG and IgM for relieving autoimmune damages in the treatment of SLE (LN).
ABSTRACT
ObjectiveTo detect the serum level of follistatin-like protein 1 (FSTL1) in patients with systemic lupus erythematosus (SLE) and its expression in renal biopsy tissues in lupus nephritis (LN) patients as well as its clinical significance were analyzed.MethodsThe serum concentration of FSTL1 in 54 SLE patients and 27 healthy controls was measured with enzyme-linked immunosorbent assay (ELISA).The distribution of FSTL1 in renal biopsy tissues was stained by immune-histochemical method.Mann-Whitney U test,t test,X2test and Pearson test were selected to compare the changes and data analysis.ResultsThe serum FSTLI level was significantly higher in SLE patients(26±21) μg/L than those of healthy controls ( 12± 14) μg/L (P＜0.01).The level of serum FSTL1 was significantly higher in SLE patients with hypertension than in patients without hypertension.The serum FSTL1 level had statistically significant changes between SLE patients with disease duration ≥ 5 years and ＜5 years.The level of serum FSTL1 correlated positively with SLEDAI score (r=0.319,P=0.022),age (r=0.700,P＜0.01),disease duration (r=0.513,P＜0.01),complement C4 level (r=0.443,P=0.004),and total serum cholesterol level (r=0.460,P=0.001 ).FSTL1 correlated inversely with platelet count (r=-0.422,P =0.001 ),anti-dsDNA antibody levels (r=-0.276,P=0.046).FSTL1 expression was evident in the cytoplasm of epithelial cells of kidney tubules.ConclusionThe level of serum FSTL1 is significantly increased in SLE patients.FSTL1 concentration correlats positively with disease activity.These data indicate that FSTL1 may play a role in the pathogenesis of SLE.