Analisis Apoptosis Jalur Ekstrinsik pada Karsinoma Sel Skuamosa Rongga Mulut Tikus (Rattus norvegicus) dengan Pemberian Nanoherbal Sikkam (Bischofia javanica) dan Mahkota Dewa (Phaleria macrocarpa)

Abstract

Oral squamous cell carcinoma (OSCC) is a common type of cancer in the head and neck region, particularly prevalent in Southeast Asia due to the consumption of tobacco with betel and areca nuts. In tobacco, Benzo(alpha)pyrene (BaP) is associated with Aryl Hydrocarbon Receptor activation, oxidative damage, and DNA mutations that trigger uncontrolled cancer cell growth. This research aims to analyze the administration of nano-herbs sikkam and mahkota dewa in combination with the expression of extrinsic apoptosis pathway proteins in OSCC rats. Nano-herb sikkam and mahkota dewa are used to induce apoptosis in OSCC. Nano-herbs were prepared using the High Energy Milling (HEM) method and analyzed using Scanning Electron Microscopy (SEM), Particle Size Analysis (PSA), phytochemical screening, characteristic testing, and toxicity testing. The OSCC rat model was established by injecting BaP at 0.04 mg/0.04 ml corn oil into the right buccal mucosa three times a week for four weeks. The study consisted of six groups: normal rats, OSCC rats, OSCC rats treated with nano-herb sikkam, OSCC rats treated with nano-herb mahkota dewa, OSCC rats treated with a combination of nano-herb sikkam and mahkota dewa, and OSCC rats treated with vitamin C. Nano-herb sikkam (800 mg/kgBW), nano-herb mahkota dewa (500 mg/kgBW), their combination, and vitamin C (40 mg/kgBW) were administered from week 6 to week 9 once daily. The ELISA technique was employed to analyze MDA, SOD, and VEGF levels in blood serum. Immunohistochemical staining was performed to analyze the expression of MDA, HSP-70, Caspase-8, Caspase-10, and Caspase-3 in cancer tissues. Results of SEM revealed that nano-herb sikkam particles had a size of 6 nm and mahkota dewa had a size of 7.7 nm, with complex surfaces and crystalline structures. PSA results showed a reduction in the size of sikkam particles from 7084.8 nm to 188.8 nm and mahkota dewa from 5062.5 nm to 246.5 nm. Qualitative phytochemical screening confirmed that both nano-herbs contained flavonoids, terpenoids, steroids, and saponins. Characteristic testing indicated that both materials met the standards of the Indonesian Herbal Pharmacopoeia for herbal drug development. Toxicity testing confirmed that both materials were categorized as mildly toxic. The combination of nano-herb mahkota dewa and sikkam significantly reduced MDA and VEGF levels, increased SOD levels, and stimulated the expression of HSP-70, Caspase-8, Caspase-10, and Caspase-3. These herbal compounds promoted apoptosis through caspase-3 with flavonoids that activated caspase-8 and caspase-10 as a caspase initiator.