Objective: The aim of this research was to investigate the potential of Sonneratia alba fruit extract to prevent atherosclerosis formation. Methods: Eighteen male Wistar rats were divided into standard or negative control (S), positive control (P) and treatment (T) groups. Atherosclerosis was induced in groups P and T by orally administering a single dose of Vitamin D3 and a high-fat diet for three days. Sonneratia alba fruit extract was given to the T-group for three days. Lipids were enzymatically measured and foam cells were counted in 10 fields of a microscopic view of the abdominal aorta. Results: This study showed higher cholesterol, LDL, HDL and triglyceride levels in the T-group, compared to the other groups. The average number of foam cells in the S-, P-and T-groups were 11.8±3.3 cells, 21.2±2.2 cells and 11.7±2.9 cells, respectively. Statistical analysis with One-Way Anova showed a significant difference in the average number of foam cells (p = 0.042). Further analysis showed a significant difference in the average number of foam cells in groups S with P (p = 0.041) and P with T (p = 0.024). Conclusion: Sonneratia alba fruit extract showed a potential effect to inhibit atherosclerosis process but could not suppress lipid levels in the blood.

Endometriosis is characterized by the presence of endometrial-like tissues located outside the uterine cavity. The expression of focal adhesion genes including FN1 gene increased in endometriosis. Epigenetic factors play a role in the alteration of expression that lead to the pathological conditions. This study aimed to analyze the mRNA expression and promoter methylation level of FN1 gene as an epigenetic mechanism in endometriosis. Forty of the total samples from endometrial patients and normal were used. The DNA and RNA were isolated, DNA was converted using sodium bisulfite procedure, amplified by MSP method. Promoter methylation level was determined by intensity of the bands that arose in gel electrophoresis using ImageJ software. The RNA was reverse transcribed into cDNA and was amplified using RT-qPCR. The mRNA expression level of FN1 gene was higher in endometrial endometriosis compared to normal, but not significant statistically (p=0,63). There was a significant difference methylation level of FN1 gene in endometrial endometriosis compared to normal endometrium (p=0.022). There was positive correlation between promoter methylation level to its mRNA expression in endometrial endometriosis (r=0.080; p=0.736). DNA methylation alteration in promoter FN1 gene was not to be caused by the increasing of mRNA expression.

Focal adhesion molecules involve in cellular migration, attachment, and play a role in endometriosis pathomechanisms. Recent studies showed that the expression of RAC1, a gene encoded focal adhesion molecule, was predominantly found in endometriosis. As gene expression may be regulated by DNA methylation. Therefore, this study aimed to analyze promoter methylation level of RAC1 gene and mRNA expression in endometrial and peritoneal endometriosis tissues. This study using 20 endometrial and 9 peritoneal tissues from the same patients and 20 normal endometrial. The DNA and RNA from samples were isolated, DNA was converted using sodium bisulfite and amplified using Methyl Specific Polymerase Chain Reaction (MSP) method. The methylation level was determined by the intensity measurement of the bands that arose in gel electrophoresis using ImageJ software, whereas mRNA expression level was measured by Reverse Transcription-quantitative PCR (RT-qPCR) method. The mRNA expression level of RAC1 gene in peritoneal endometriosis increased compared to normal endometrium, as well as compared to endometrial endometriosis, but there was no significant difference in endometrial endometriosis compared to normal. Promoter hypermethylation level of RAC1 gene in peritoneal endometriosis was significantly different compared to normal endometrium, however not significant to endometrial endometriosis. Methylation level of its gene in endometrial endometriosis shown no significant difference compared to normal. There was association between promoter hypermethylation level and its mRNA expression in endometrial endometriosis (R= 0.014; p=0.952). The elevation of mRNA expression of RAC1 gene plays a role in endometrial cell migration to peritoneum, and associated with promoter hypermethylation level of its gene.

Endometrial cancer (EC) rates are rising annually. Additional prediction markers need to be evaluated because only 10–20% of EC cases show an objective response to immune-checkpoint inhibitors (ICIs). Our previous methylomic study found that BHLHE22 is hypermethylated in EC tissues and can be detected using a Pap-smear sample. BHLHE22, a basic helix loop helix transcription factor family member, is known as a transcriptional repressor and is involved in cell differentiation. However, the role of BHLHE22 in EC remains poorly understood. Herein, we analyzed BHLHE22 expression in 54 paired cancer and normal endometrial tissue samples, and confirmed with databases (TCGA, GTEx, and human protein atlas). We found that BHLHE22 protein expression was significantly downregulated in EC compared with normal endometrium. High BHLHE22 expression was associated with microsatellite-instable subtype, endometrioid type, grade, and age. It showed a significant favorable survival. BHLHE22 overexpression inhibited the proliferation and migration of EC cells. Functional enrichment analysis showed that BHLHE22 was significantly associated with immune-related pathways. Furthermore, BHLHE22 was positively correlated with proinflammatory leukocyte infiltration and expression of chemokine genes in EC. In conclusion, BHLHE22 regulates immune-related pathways and modulates the immune microenvironment of EC.

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