ABSTRACT
We aimed to investigate therapeutic effect of Bushenhuoxue recipe in intrauterine adhesions (IUA) and explore the underlying molecular mechanism via integrating network pharmacology and in vitro experimental verification. The active compounds and gene targets of Bushenhuoxue recipe were screened in the TCMSP database and the IUA-related genes were identified using GeneCards database by the keyword "Intrauterine adhesions". Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were conducted to reveal the underlying molecular mechanism of Bushenhuoxue recipe treating IUA. T-HESC cells were inducted to fibrotic state using TGF-ß1 of 10 ng/ml concentration treating for 24 h. RT-qPCR or western blot was used to demonstrate the expression levels of fibrosis markers (COL1A1 and α-SMA) and KEGG pathway markers. Cell counting kit-8 (CCK8) assay was performed to illustrate the cell viability of endometrial stromal cell. The treatment of Bushenhuoxue recipe could significantly inhibit the proliferation and fibrosis of endometrial stromal cells. We obtained a total of 169 no-repeat ingredients of Bushenhuoxue recipe and 3044 corresponding targets. After taking intersection with 4230 no-repeat IUA-related genes, a total of 83 target genes related to both Bushenhuoxue recipe and IUA were finally identified. KEGG analysis found that PI3K-AKT signaling pathway might be the key pathway. Further experiment revealed that PI3K-AKT signaling pathway was significantly activated in endometrial stromal cells of fibrotic state and the treatment of Bushenhuoxue recipe could inhibit the PI3K-AKT signaling pathway. Further rescue assay demonstrated that Bushenhuoxue recipe suppressed the proliferation and fibrosis of endometrial stromal cells via PI3K-AKT signaling pathway. Bushenhuoxue recipe suppresses the proliferation and fibrosis of endometrial stromal cells via PI3K-AKT signaling pathway, eventually inhibiting the progression of IUA.
ABSTRACT
BACKGROUND: To investigate impacts of sodium arsenic (NaAsO2) on embryonic cardiac development in rats and evaluate the protective role of folate in NaAsO2 exposure rats. METHODS: We divided 90 female rats randomly into 9 groups. Group A was the control; group B-F were the animals fed with NaAsO2 in a series of increased doses, corresponding to 9.4 mg/L, 18.8 mg/L, 37.5 mg/L, 75 mg/L and 150 mg/L, respectively; group G-I were fed with 75 mg/L of NaAsO2, in addition of folate with doses of 0.53 mg/kg, 5.3 mg/kg, and 10.6 mg/kg, respectively. Their fetus' general development and cardiovascular systems were examined. Nkx2.5, GATA4, TBX5 gene and protein expression were measured. RESULTS: Relatively to group A, arsenic treated group C-F rats generated significantly lower weight of fetus and placenta (P<0.05), whereas the folate-treated groups H and I were significantly heavier than the arsenic-treated group E (P<0.05). We observed that incidences of cardiac malformations were significantly greater in arsenic-treated group E and F than group A (P<0.05). We found that the Nkx2.5 and GATA4 protein expression in the fetal hearts were downregulated in group B-F compared to group A. But the expression of them was significantly upregulated in group H-I relatively to group E (P<0.05). Moreover, the TBX5 gene expression was increased in both group D-F and G-I when they were compared to group A or group E, respectively (P<0.05). CONCLUSION: NaAsO2 induce embryonic cardiac defection and folate supplement alleviate this impairment through modulation of the Nkx2.5, GATA4 and TBX5 gene expression.
ABSTRACT
OBJECTIVE: To investigate the effects of arsenic exposure before and during maternal pregnancy on heart development of fetal rats. METHODS: According to body weight, thirty-two female SD rats (30 to 40 days of age) were randomly divided into control group, low dose group, middle dose group and high dose group with 8 rats per group. They were allowed free access to drinking water with 0, 37.5, 75 and 150 mg/L of sodium arsenite (NaAsO2) for 6 weeks, respectively. Then all the female rats and adult male SD rats were caged together for mating. Once female rats were determined to be pregnant, they would continue to drink deionized distilled water containing different concentrations of sodium arsenite for another 2 weeks. On embryonic day 16, rats were sacrificed to harvest fetuses. Female rats' weight changes, abortions, absorbed fetus number, growth and development of fetal rats were observed. Hematoxylin-eosin staining of serial cardiac slices was performed in embryos to observe cardiac morphology and structure. Fur arsenic contents of female rats were determined with the method of atomic fluorescence spectrometry. RESULTS: Subchronic arsenic exposure caused slow weight growth in female rats. There were two cases of abortion in middle dose group and high dose group, respectively. Compared with these of control group, fetal and placental weight decreased (P < 0.05), and the incidence of fetal absorption increased (P < 0.05) in all arsenic-treated groups. Cardiac malformations in fetal rats including ventricular septal defect, atrial septal defect and tetralogy of Fallot were observed in low, middle and high dose group. The incidence of cardiac malformations increased with the increase of arsenic concentrations in drinking water. Compared with that of control group, the incidence of cardiac malformations remarkably increased in both middle and high dose groups (P < 0.05). Fur arsenic contents increased with the increase of arsenic concentrations in drinking water (P < 0.01). CONCLUSION: Arsenic exposure before and during maternal pregnancy could cause abnormal cardiac development in fetal rats, and increased the risk of congenital heart disease.
