ABSTRACT
Objective:To analyze the material basis, target and pathway of Chebulae fructus and Margarita in Mongolian medicine Garidi-13 Pill on stroke with the method of network pharmacology and molecular docking technology, so as to better understand its "detoxification" mechanism. Methods:TCMIP and BATMAN-TCM were used to predict the target of Chebulae fructus and Margarita composition, and GeneCards were used to search for the target of stroke. The overlapping targets of the two platforms were imported into the Metascape software for GO biological analysis and KEGG pathway analysis. The molecular docking of key molecules and targets was carried out with LEDOCK. Results:A total of 22 active components were collected and 217 targets related to stroke were predicted. Among them, 1-O-galloyl-glucose, cuprum, ellagicacid, arjungenin and corilagin were the key substances playing the role of "detoxification" of the Chebulae fructus and Margarita; IL6, TNF, HSP90AA1, PTGS2, CASP3, NR1I2, VKORC1 and ATP1A1 were the key targets playing the role of "detoxification" . These targets were significantly enriched in cell response, humoral level regulation, hemostasis, response to steroid hormones, steroid metabolism, coagulation and other biological processes, as well as nitrogen metabolism, IL-17 signaling pathway and other pathways. Molecular docking verified the accuracy of previous prediction results from computer simulation level. Conclusion:The process of Chebulae fructus and Margarita intervening strok is closely related to the elimination of harmful metabolites and calmingthe inflammatory reaction, which was not only consistent with the modern medicine on the pathological process of stroke, but also consistent with the interpretation of "evil and poison" with Mongolian medicine theory.
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Objective:To investigate the effect of miR-129-3p on the proliferation, migration and invasion of hepatocellular carcinoma cells by targeting E2F transcription factor 5 (E2F5).Methods:The expression of miR-129-3p and E2F5 in different liver cancer cell lines and normal liver cell lines were detected by fluorescence quantitative polymerase chain reaction. HepG2 cell lines overexpressing miR-129-3p were constructed, thiazole blue cell proliferation assay and plate cloning test were used to detect the proliferation and clone formation ability of each cell. Cell scratch assay and transwell assay were used to detect cell migration and invasion. Western blotting was used to detect the protein expression. The targeting relationship between miR-129-3p and E2F5 was verified by dual luciferase reporter gene method and western blotting. The experimental groups were as follows: non-transfection (NC) group; transfection control miR-con (miR-con) group; transfection of miR-129-3p (miR-129-3p) group; co-transfection of miR-129-3p and empty vector pcDNA (miR-129-3p+ pcDNA) group; co-transfection of miR-129-3p and pcDNA-E2F5 (miR-129-3p+ pcDNA-E2F5) group.Results:The expression of miR-129-3p in MHCC-97H, HepG2, LM3, Hep3B, PLC, HUH7 were all lower than that of HL-7702, the difference were statistically significant (all P<0.05). The number of cell clones [(86.56±20.84) vs. (511.29±45.03)and(509.78±40.81)], the cell migration rate [(3.03±1.29)% vs. (15.01±2.30)% and(14.99±2.31)%], the relative expression of migration-related protein MMP-2 [(0.51±0.22)vs.(1.87±0.30)and(1.84±0.35)]and the number of transmembrane cells [(33.10±1.58) vs. (101.23±0.31) and (100.96±3.44)] in miR-129-3p group were all decreased when compared with NC group and miR-Con group, the difference was statistically significant ( P<0.05). Dual luciferase reporting assay showed that miR-129-3p can negatively target E2F5 and inhibit its expression. Conclusion:The expression level of miR-129-3p is low in hepatocellular carcinoma cells, overexpression of miR-129-3p can inhibit the malignant biological behavior of hepatocellular carcinoma cells by targeting E2F5.
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Objective To determine the level of peripheral CD4+CD25+ regulatory T lymphocytes in patients with vitiligo at different stages and to study its relationship with the development of vitiligo. Methods Blood samples were collected from 34 outpatients with vitiligo, including 19 cases of progressive vitiligo and 15 cases of stable vitiligo, as well as from 20 normal human controls. Flow cytometry was used to detect the levels of peripheral CD4+ and CD4+CD25+ T lymphocytes in these samples. Results Compared with the controls, the percentage of CD4+CD25+ regulatory T lymphoeytes in peripheral lymphocytes was significantly lower in patients with progressive vitiligo than those in patients with stable vitiligo and normal human con-trois [(2.43±0.30)% vs (3.49±0.39)% and (3.34±0.24)%, both P <0.05], but no significant difference was found between patients with stable vitiligo and normal human controls (P＞0.05). A significantly nega-tive correlation was observed between the percentage of CD4+CD25+ regulatory T lymphocytes and lesion area in patients with progressive vitiligo (r = -0.48, P <0.05), but not in patients with stable vitiligo (P ＞0.05). There was no significant correlation between the course of disease and the percentage of peripheral CD4+CD25+ regulatory T lymphocytes in patients with progressive vitiligo or stable vitiligo (both P ＞ 0.05). Conclusion There is an abnormal proportion of peripheral CD4+CD25+ regulatory T lymphocytes in patients with vitiligo, which may be related to the development of vitiligo.