Tesis

Respons Hipoksia Makrofag pada Pasien Tuberkulosis Paru Resisten Obat dan Kontak Erat: Kajian terhadap Ekspresi HIF-1α, HIF- 2α, dan Sitoglobin = Hypoxic Response of Macrophages in DrugResistant Tuberculosis Patients and Closed Contact: Study of HIF-1α, HIF-2α, and Cytoglobin Expressions.

Tuberkulosis masih menjadi masalah kesehatan utama karena prevalensinya yang terus meningkat, terutama kasus TB resisten obat. Penelitian ini bertujuan untuk menganalisis respons adaptasi makrofag terhadap hipoksia dan kemampuan fagositosis makrofag pada penderita TB resisten obat dibandingkan dengan kontak erat yang terinfeksi laten dan sehat. Enam pasien TBRO dan 18 kasus kontak erat (8 TB laten; 10 sehat) di RS Universitas Indonesia direkrut sebagai subjek penelitian. Makrofag berasal dari hasil isolasi sel mononukleus darah tepi (SMDT) subjek yang dikultur selama 7 hari. Pemeriksaan ekspresi mRNA dan protein HIF- 1α, HIF-2α, CYGB dilakukan menggunakan qRT-PCR dan ELISA. Hasil menunjukkan bahwa aktivitas fagositosis kelompok infeksi laten lebih tinggi dibandingkan kelompok sehat dan TB RO (p < 0,05). Ekspresi mRNA dan protein HIF-1α, HIF-2 α, dan CYGB lebih tinggi pada kelompok TB RO dibandingkan kelompok lainnya (p < 0,05). Korelasi positif didapatkan antara ekspresi mRNA dengan protein HIF-1α, HIF-2α, dan CYGB, serta protein HIF-1α dengan mRNA CYGB (r = 0,517; r = 0,697; r = 0,890; r = 0,763; p < 0,05). Terdapat korelasi positif kuat pada ekspresi antar protein regulator hipoksia (HIF-1α dengan HIF-2α dan CYGB; HIF-2α dengan CYGB dengan nilai r = 0,926; r = 0,921; 0,884; p < 0,001; Pearson correlation). Namun terdapat korelasi negatif sedang antara kemampuan fagositosis dengan ekspresi protein HIF-1α, HIF-2α, dan CYGB (r = -0,612; r = - 0,599; r= -0,512; p < 0,05; Pearson correlation). Perbedaan respons adaptasi hipoksia dan fungsi sel makrofag diharapkan dapat menjadi referensi selanjutnya dalam membuka penelitian yang lebih spesifik, untuk menelusuri lebih lanjut dari aspek lain mengenai respons imun makrofag pada penderita TB resisten obat dibandingkan dengan kontak erat terdiagnosis laten TB, dan kontak erat yang sehat.
Kata Kunci: Hipoksia, Makrofag, Tuberkulosis Resisten Obat


Tuberculosis remains a major health problem due to its continuously increasing prevalence, particularly in cases ofdrug-resistant TB. This research aims to analyze the macrophage adaptive response to hypoxia and the phagocytic ability of macrophages in drug-resistant TB patients compared to latently infected and healthy close contacts. Six drug-resistant TB patients and 18 close contact cases (8 latent TB; 10 healthy) at the University of Indonesia Hospital were recruited as study subjects. Macrophages were derived from peripheral blood mononuclear cell isolation from subjects and cultured for 7 days. The mRNA and protein expression ofHIF-1α, HIF-2α, and CYGB were examined using qRT-PCR and ELISA. Results showed that the phagocytic activity of the latent infection group was higher than the healthy and drug-resistant TB groups (p < 0.05). The mRNA and protein expression of HIF-1α, HIF-2α, and CYGB were higher in the drug-resistant TB group compared to the other groups (p < 0.05). Positive correlations were found between mRNA expression and protein levels ofHIF-1α, HIF-2α, CYGB, and HIF- 1α protein with CYGB mRNA (r = 0.517; r = 0.697; r = 0.890; r = 0.763; p < 0.05). There was a strong positive correlation in the expression of hypoxia regulator proteins (HIF-1α with HIF-2α and CYGB; HIF-2α with CYGB with r = 0.926; r = 0.921; 0.884; p < 0.001; Pearson correlation). However, a moderate negative correlation was observed between phagocytic ability and the expression of HIF-1α, HIF-2α, and CYGB proteins (r = -0.612; r = -0.599; r= -0.512; p < 0.05; Pearson correlation). The differences in hypoxia adaptation response and macrophage cell function are expected to serve as a reference for more specific research, delving into other aspects of macrophage immune response in drug-resistant TB patients compared to close contacts with latent TB diagnosis and healthy close contacts.
Keywords: Hypoxia, Macrophage, Drug-Resistant Tuberculosis