Endometriosis merupakan penyakit ginekologi yang sering dikaitkan dengan nyeri panggul pada subjek usia reproduksi. Namun, etiologi penyakit ini sangatlah kompleks, yang melibatkan faktor epigenetik. Metilasi DNA meregulasi perubahan ekspresi gen yang menyebabkan disregulasi hormone estrogen pada sel endometriosis. Diketahui gen NGF dan P2RX3 mengalami perubahan metilasi DNA dan peningkatan ekspresi mRNA pada peritoneum endometriosis dan berkorelasi terhadap intensitas nyeri endometriosis. Penelitian ini bertujuan untuk mengukur metilasi DNA, ekspresi mRNA dan analisis korelasinya terhadap gen NGF dan P2RX3 pada darah menstruasi subjek endometriosis dan tanpa endometriosis, serta menganalisis korelasi antara darah menstruasi dengan endometrium eutopiknya pada masing-masing variabel. Pengukuran metilasi DNA gen NGF dan P2RX3 menggunakan metode Methylated Specific PCR (MSP) dan pyrosequencing. DNA genom setiap sampel jaringan diekstraksi, dan dimodifikasi bisulfit. Perhitungan metilasi DNA metode MSP berdasarkan pengukuran intensitas pita yang terukur oleh software ImageJ, sedangkan metode pyrosequencing dihitung berdasarkan perhitungan nilai rata-rata dari semua lokasi CpG yang dianalisis. Pengukuran ekspresi mRNA menggunakan RT-qPCR dan dianalisis dengan metode Livak. Hasil pengukuran metilasi DNA gen NGF mengalami hipometilasi sebesar 39,3% (p=0,025); gen P2RX3 lebih termetilasi 81,1% (p=0,799) pada darah menstruasi subjek endometriosis dibandingkan dengan kelompok tanpa endometriosis. Terdapat peningkatan ekspresi mRNA NGF secara bermakna sebesar 4,29 kali (p=0,021); ekspresi mRNA P2RX3 lebih tinggi 2,23 kali (p=0,004) pada darah menstruasi subjek endometriosis dibandingkan terhadap tanpa endometriosis. Penelitian ini tidak menunjukkan korelasi yang bermakna antara metilasi DNA dan ekspresi mRNA baik gen NGF maupun P2RX3 di darah menstruasi subjek endometriosis (Gen NGF nilai p=0,702; r=-0,179, dan gen P2RX3 nilai p=0,461; r=0,264). Namun, analisis korelasi antara darah menstruasi dan endometrium eutopik subjek endometriosis menghasilkan korelasi yang positif, baik pada gen NGF maupun P2RX3. Hipometilasi gen NGF pada darah menstruasi dapat dijadikan biomarker dalam pengembangan diagnostik endometriosis non-invasif.

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Endometriosis is a gynecological disease that is often associated with pelvic pain in women of reproductive age. However, the etiology of this disease is complex, involving epigenetic factors. DNA methylation regulates gene expression changes that cause dysregulation of the hormone estrogen in endometrial cells. It is known that the NGF and P2RX3 genes have altered DNA methylation and increased mRNA expression in the endometriosis peritoneum and correlated with the intensity of endometriosis pain. This study aims to measure DNA methylation, mRNA expression and analysis of its correlation to the NGF and P2RX3 genes in menstrual blood of subjects with endometriosis and without endometriosis, as well as analyze the correlation between menstrual blood and its eutopic endometrium in each variable. Measurement of DNA methylation of NGF and P2RX3 genes using Methylated Specific PCR (MSP) and pyrosequencing methods. The genomic DNA of each tissue sample was extracted, and bisulfite modified. The calculation of DNA methylation using the MSP method is based on the measurement of the band intensity measured by the ImageJ software, while the pyrosequencing method is calculated based on the calculation of the average value of all analyzed CpG locations. Measurement of mRNA expression using RT-qPCR and analyzed by the Livak method. The results of DNA methylation measurements of the NGF gene were hypomethylated by 39.3% (p=0.025); P2RX3 gene was more methylated 81.1% (p=0.799) in menstrual blood of endometriosis subjects compared to the group without endometriosis. There was a significant increase in NGF mRNA expression by 4.29 times (p=0.021); P2RX3 mRNA expression was 2.23 times higher (p=0.004) in menstrual blood of endometriosis subjects compared to those without endometriosis. This study did not show a significant correlation between DNA methylation and mRNA expression of both NGF and P2RX3 genes in menstrual blood of endometriosis subjects (NGF gene p value = 0.702; r = -0.179, and P2RX3 gene p value = 0.461; r = 0.264). However, correlation analysis between menstrual blood and eutopic endometrium of endometriosis subjects yielded a positive correlation, both in the NGF and P2RX3 genes. Hypomethylation of the NGF gene in menstrual blood can be used as a biomarker in the development of non-invasive diagnostic endometriosis.