ABSTRACT
OBJECTIVE： To predict the anti-inflammatory active components and mechanism of couplet medicine of Notopterygium incisum-Angelica pubescens. METHODS： According to the principle of oral bioavailability≥30％ and drug- likeness≥0.18， active components of N. incisum and A. pubescens were screened； TCMSP was used to predict and screen the potential target of them. Using “Anti-inflammatory” as keyword， inflammatory related target genes were retrieved from human gene database Genecards. Common target was screened by mapping the target genes of active ingredients from couplet medicine of N. incisum-A. pubescens. The active ingredient-target network was established by using Cytoscape 3.5.1 software. The screened targets were used to construct the target protein interaction （PPI） network on the STRING V 10.5 platform. Its anti-inflammatory mechanism was studied by KEGG signaling pathway and GO biological enrichment analysis. RESULTS： Totally 15 active components such as coumarin， beta-sitosterol， ammidin， nodakenin were selected from couplet medicine of N. incisum-A. pubescens. Acting on 49 targets such as transcription factor AP-1， PI3-kinase subunit gamma， estrogen receptor， they mainly involved 19 signaling pathways such as hepatitis B and cell apoptosis， and were involved in 47 biological processes such as regulating inflammatory response and prostaglandin biosynthesis. CONCLUSIONS： The anti-inflammatory mechanism of active components of couplet medicine of N. incisum-A. pubescens on multi-target， multi-channel and multi-biological processes is predicted， and it points out the direction for further anti-inflammatory mechanism study.
ABSTRACT
Objective To investigate the functions of microRNA-1 43 (miR-1 43)in esophageal cancer cell line ECA1 09.Methods ECA1 09 cells were transfected with negative control (NC),miR-1 43 mimics or miR-1 43 inhibitors.3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT)assay was per-formed to evaluate the growth of ECA1 09 cells after transfection.Annexin V-FITC /PI apoptosis test kit was used to detect early apoptosis rate in ECA1 09 cells.Transwell migration and invasion assays were conducted to compare the migration and invasion capacity of ECA1 09 among different groups.Real-time PCR and Western blotting were used to analyze the mRNA and protein alteration after transfection.Results Three and four days after transfection,compared with NC (absorbance value:0.90 ±0.02 and 1 .09 ±0.07),miR-1 43 mimics inhibited ECA1 09 cell proliferation (absorbance value:0.66 ±0.05 and 0.80 ±0.04),while miR-1 43 inhibi-tors promoted cell proliferation (absorbance value:1 .1 3 ±0.09 and 1 .51 ±0.08),with statistical signifi-cances (F =49.1 6,P =0.000;F =1 00.34,P =0.000).Early-stage apoptosis rates of ECA1 09 transfected with NC,miR-1 43 mimics and miR-1 43 inhibitors were 3.42% ±0.72%,1 1 .63% ±1 .1 5% and 0.94% ± 0.1 0%,respectively,with statistical significance (F =1 51 .61 ,P =0.000).Meanwhile,compared with NC (migration cell number:336 ±1 3,invasion cell number:1 47 ±1 6),miR-1 43 mimics inhibited cell migration (1 48 ±1 6)and invasion (75 ±1 0),while miR-1 43 inhibitors promoted cell migration (51 0 ±1 4)and inva-sion (238 ±1 6),with statistical significances (F =470.99,P =0.000;F =90.04,P =0.000).Compared with NC (1 .00 ±0.00),miR-1 43 mimics down-regulated mRNA (relative expression level 0.22 ±0.08)and protein expression (relative expression level 0.46 ±0.08)of K-ras,whereas miR-1 43 inhibitors up-regulated mRNA (1 .55 ±0.1 2)and protein expression (1 .33 ±0.05)of K-ras (F =1 31 .36,P =0.000;F =88.1 7, P =0.000).Conclusion miR-1 43 functions as a tumor suppressor in esophageal cancer cell line ECA1 09, probably by down-regulating K-ras expression.