ABSTRACT
OBJECTIVE Forsythiae Fructus (Lianqiao) is a typical heat-clearing and detoxicating traditional Chinese medicine (TCM) herb, which has been traditionally used for treating cancer according to TCM theory. However, the under?lying mechanism has not been fully explained. METHODS In this study, we investigated the antitumor effect of Forsyth?iae Fructus aqueous extract (FAE) on B16-F10 melanoma. RESULTS FAE strongly inhibited the tumor growth and metastasis formation in B16-F10 melanoma transplanted mice. The survival time of tumor-bearing mice was also signifi?cantly prolonged by FAE. The levels of ROS, MDA, TNF-α and IL-6 decreased, while GSH increased in the FAE treat?ment group, indicating FAE possesses strong anti-oxidative and anti-inflammatory activity. Western blotting analysis demonstrated that antioxidant proteins Nrf2 and HO-1, tumor suppressors P53 and p-PTEN, and the MAPK pathways in tumor tissues were upregulated by FAE treatment. Serum metabolomics analysis further uncovered that 17 metabolites mostly involving in glycerophospholipid metabolism were correlated with the antitumor effect of FAE. Notably, several lysophosphatidylcholines (LysoPCs) significantly decreased in tumor model group, while FAE treatment restored the changes of these phospholipids to about normal condition. LysoPC acyltransferase 1 (LPCAT1) and autotaxin (ATX) highly expressed in melanoma and markedly downregulated by FAE were believed to be responsible for this modulation. CONCLUSION FAE exhibites strong antitumor activity against B16-F10 melanoma through activating MAPKs/Nrf2/HO-1 mediated anti-oxidation and anti-inflammation and modulating glycerophospholipid metabolism via downregulating LPCAT1 and ATX. Besides, it is suggested that serum LysoPCs could be potential biomarkers for the diagnosis and prog?nosis of melanoma.
ABSTRACT
<p><b>OBJECTIVE</b>To explore different microRNA expression profiles between chronic hepatitis B (CHB) patients of Pi-Wei dampness-heat syndrome (PWDHS) and Gan depression Pi deficiency syndrome (GDPDS).</p><p><b>METHODS</b>By applying gene chip technology, blood samples from CHB patients of PWDHS (3 cases), GDPDS (3 cases), and healthy volunteers (3 cases) were withdrawn and microRNA detected. The microRNA was screened and functional analyses performed by using SAS system.</p><p><b>RESULTS</b>Totally 77 microRNAs with differential expression were screened from CHB patients of PWDHS and healthy volunteers, including 60 up-regulated microRNAs and 17 down-regulated microRNAs. Functions of target genes were mainly associated with transcription factors, gas exchange, adverse stimulating, regulation of enzyme activities, developing of the immune system, and the process of actin filaments. Totally 41 microRNAs with differential expression were screened from CHB patients of GDPDS and healthy volunteers, including 32 up-regulated microRNAs and 9 down-regulated microRNAs. Functions of target genes were mainly associated with binding to nucleotide or chromatin, inhibition and activation of transcription, biosynthesis, regulation of metabolic process, regulation of enzyme activities, developing of the immune system, the process of actin filaments, and IL-12. Totally 6 microRNAs with differential expression were screened from CHB patients of PWDHS and CHB patients of GDPDS, including 1 up-regulated microRNA and 5 down-regulated microRNAs. Functions of target genes were mainly associated with transmembrane transport, regulation of transcription factors, metabolism of hormones, developing of the immune system, the process of actin filaments, regulation of metabolic process, response to exterior stimulation, and so on.</p><p><b>CONCLUSION</b>There existed differentially expressed microRNAs (spectrum) between CHB patients of PWDHS and CHB patients of GDPDS.</p>
Subject(s)
Humans , Depression , Hepatitis B, Chronic , Genetics , Metabolism , Hot Temperature , Interleukin-12 , Metabolism , Medicine, Chinese Traditional , MicroRNAs , Metabolism , Oligonucleotide Array Sequence Analysis , Research , SyndromeABSTRACT
AIM: To investigate the effects of tranfection of IL-2R antisense RNA expression plasmids on mouse spleen cells' proliferation in vitro and its possible mechanism. METHODS: Spleen cells were transfected with IL-2R antisense RNA eukaryotic expression plasmids using adhesion-assisted lipofection method, and then the spleen cells were stimulated by mitogen. Cells' proliferation was tested by tetrazolium salt (MTT) method. IL-2R mRNA and protein expression level were measured by slot-blot hybridization assay and flow cytometry method respectively. RESULTS: The proliferation of spleen cells was inhibited obviously after transfecting with recombinant plasmids. The inhibitory rate of pcAnti-mlL-2Rαβ- and pciAnti-mIL-2Rαβ-transfected group was higher than that of pcAnti-mIL-2Rα-and pcAnti-mIL-2Rβ-transfected group; the inhibitory rate of pcAnti-mIL-2Rα-tranfected group was higher than that of pcAnti-mlL-2Rβ-tranfected group. No inhibitory effect on the growth of NIH3T3 cells was observed when they were transfected with recombinant plasmids. IL-2R mRNA and protein expression level were decreased in spleen cells after transfection of recombinant plasmids. CONCLUSION: IL-2R antisense RNA can efficiently inhibit the proliferation of mouse spleen cells in vitro. IL-2Rαβ chimeric antisense RNA showed higher inhibitory rate than a or antisense RNA. IL-2Rα antisense RNA was more effective than β antisense RNA. It can be concluded preliminarily that the inhibitory effect of IL-2R antisense RNA was exclusively on the growth of cells functionally expressing IL-2R. The inhibitory effect on the spleen cells' proliferation was likely due to the blocking of IL-2R expression by antisense RNA.