Disertasi

Potensi dan Mekanisme Antifibrotik Nanosuspensi Kurkumin Inhalasi dalam Kombinasi dengan Kortikosteroid pada Model Tikus Fibrosis Paru Idiopatik dengan Eksaserbasi Akut. = Antifibrotic Effects and Mechanism of Curcumin Nanosuspension for Inhalation in Combination with Corticosteroids in the Rat Model of Acute Exacerbations of Idiopathic Pulmonary Fibrosis.

Latar belakang: Fibrosis paru idiopatik dengan eksaserbasi akut (AE-IPF) merupakan kejadian yang mengancam jiwa, dengan tingkat mortalitas tinggi, namun hingga sekarang tidak terdapat obat yang dapat mengatasi masalah ini. Kurkumin diketahui memiliki berbagai efek farmakologi termasuk antiinflamasi dan antifibrotik, namum memiliki masalah dalam solubilitas dan distribusi ke organ target. Pada penelitian ini, sediaan nanosuspensi kurkumin inhalasi diformulasikan untuk diberikan dan dibuktikannya efeknya kepada model hewan AE-IPF. Metode: Nanosuspensi kurkumin (NSK) dibuat dengan kombinasi metode anti solvent precipitation dan high speed homogenization. NSK dikarakterisasi ukuran partikel, indeks polidispersitas (PDI), zeta potensial dan efisiensi penjerapannya. NSK kemudian dibuktikan efikasinya pada tikus Sprague-Dawley yang diinduksi bleomisin (BLM) 5 mg/kg intratrakeal dan lipopolisakarida (LPS) 7,5 mg/kg intraperitoneal. Tikus sebanyak 24 ekor dibagi menjadi 6 kelompok, yaitu (1) kelompok normal/sham, (2) kelompok BLM-LPS, (3) kelompok BLM-LPS + prednison 5 mg/kg per oral, (4) kelompok BLM-LPS + prednison 5 mg/kg per oral dan NSK 1 mg/kg inhalasi, (5) kelompok BLM-LPS + prednison 5 mg/kg per oral dan NSK 10 mg/kg inhalasi, dan (6) kelompok BLM-LPS + NSK 10 mg/kg inhalasi.Terapi dilakukan selama 7 hari. Setelah itu dilakukan pengukuran saturasi oksigen (SpO2), laktat dehydrogenase (LDH), indeks paru, profil histopatologi paru, kadar hidroksiprolin, ekspresi α-smooth muscle actin (α-SMA), kadar Transforming Growth Factor beta 1 (TGFβ-1), Ashcroft modified score, area densitas kolagen, area udara alveolar, hitung jenis sel bilasan bronkoalveolar (BALF), ekspresi IL-6 dan IL-8 serta kadar angiotensin II, Angiotensin Converting Enzyme 2 (ACE2) dan ekspresi Angiotensin II type 1 Receptor (AT1R). Hasil: Nanosuspensi kurkumin berhasil dibuat dengan ukuran partikel 281,3 nm, PDI 0,464, zeta potensial -29,4 mv dan efisiensi penjerapan 90,11%. Kombinasi nanosuspensi kurkumin dengan prednison telah dibuktikan efikasinya pada hewan model AE-IPF, dengan peningkatan SpO2, penurunan indeks paru, Ashcroft modified score, area densitas kolagen dan peningkatan area udara alveolar. Perbaikan pada paru tersebut diikuti peningkatan kadar ACE2, penurunan kadar IL- 6, IL-8, LDH, Angiotensin II, TGFβ-1, ekspresi protein α-SMA dan AT1R. Kesimpulan: Pemberian terapi kombinasi prednison dan nanosuspensi kurkumin inhalasi pada tikus AE-IPF dapat memberikan perbaikan fungsi dan kondisi fibrosis paru melalui perbaikan inflamasi dan peningkatan ekspresi AT1R, TGFβ-1 dan α- SMA.
Kata kunci: α-smooth muscle actin, angiotensin II, bleomisin, cairan bilasan bronkoalveolar, fibrosis, TGFβ-1.



Introduction: Acute exacerbation of idiopathic pulmonary fibrosis (AE-IPF) is a life-threatening condition with a high mortality rate and no standard treatment. Curcumin is known to have various pharmacological effects, including anti- inflammatory and antifibrotic activities; however, it has poor solubility and distribution to target organs. Thus, the present study aimed to prepare a formulation of inhaled curcumin nanosuspension and demonstrate its effect on the animal model of AE-IPF. Methods: Anti-solvent precipitation and high-speed homogenization methods were used to prepare curcumin nanosuspension (NSK) for inhalation. NSK was characterized by particle size, polydispersity index (PDI), zeta potential and entrapment efficiency. The efficacy of NSK was then established on Sprague- Dawley rats induced by intratracheal bleomycin (BLM) 5 mg/kg and intraperitoneal lipopolysaccharide (LPS) 7.5 mg/kg. Twenty-four rats were divided into six groups (1) the normal/sham group, (2) the BLM-LPS group, (3) the BLM-LPS + prednisone 5 mg/kg per oral, (4) the BLM-LPS + prednisone 5 mg/kg per oral and NSK 1 mg/kg per inhalation, (5) the BLM-LPS + prednisone 5 mg/kg per oral and NSK 10 mg/kg per inhalation, and (6) the BLM-LPS + NSK 10 mg group /kg per inhalation. The treatments were carried out for seven days. Afterward, measurements of lung function markers (oxygen saturation (SpO2), lactate dehydrogenase (LDH), lung index, lung histopathological profile, hydroxyproline, Ashcroft modified score, collagen density area, alveolar air area, bronchoalveolar lavage fluid (BALF) cell count), levels of Transforming Growth Factor beta 1 (TGFβ-1), IL-6, IL-8, angiotensin II, Angiotensin Converting Enzyme 2 (ACE2), the expressions of α-smooth muscle actin (α-SMA) and Angiotensin II type 1 Receptor (AT1R) were done. Results: Nanosuspension of curcumin was successfully prepared with a particle size of 281.3 nm, PDI of 0.464, a zeta potential of -29.4 mv, and an entrapment efficiency of 90.11%. The combination of curcumin nanosuspension with prednisone was shown to be efficacious in the AE-IPF animal model, as indicated by the improvement of SpO2 as well as decreased lung index, Ashcroft modified score, area of collagen density, and increased area of alveolar air. The improvement in lung functions was followed by increased levels of ACE2, decreased levels of IL-6, IL-8, LDH, Angiotensin II, TGFβ-1, and expression of α-SMA and AT1R. Conclusion: The combination of prednisone and curcumin nanosuspension for inhalation in animal models of AE-IPF can improve lung function and pulmonary fibrosis by ameliorating inflammation and enhancing the expression of AT1R, TGFβ-1, and α-SMA. Keywords: α-smooth muscle actin, angiotensin II, bleomycin, bronchoalveolar lavage fluid, fibrosis, TGFβ-1.