Pengaruh Pemberian Ekstrak Phyllanthus niruri Linn Terhadap Infiltrasi Limfosit dan Ekspresi Perforin pada Kanker Kolon Tikus Sprague-Dawley

Endang Sawitri, Ign. Riwanto, Tjahjono Tjahjono, Edi Dharmana


Abstract


ABSTRACT
The effect of Phyllanthus niruri Linn extract on lymphocytes infiltration and perforin expression in colon cancer of Sprague-Dawley RatBackground: Colon cancer treatment currently involves immunotherapy that aims to improve the quality of life and survival of patients. Phyllanthus niruri Linn (P. niruri L) may act as an immunomodulator, but its potency in antitumor immune responses has not been revealed. The study was conducted to evaluate the differences between the immunological status of rats suffering colon cancer which were not given to those given the extract of P. niruri L.
Methods: The study was randomized posttest-only control group design. Samples were 30 Sprague-Dawley male rats, bodyweight 170-220 gr which induced by 1.2 DMH 30 mg/kgBW subcutaneously. On the weeks 9, 11 and 13, four induced rats each week were sacrified to detect the development of colon cancer. On the weeks of 13th all of 4 rats were developed colon cancer, so the induction were stopped. The rest of 18 induced rats were randomly into two groups: without P.  niruri L or positive control (K+)=9 rats and  given P. niruri L extract 13.5 mg/kg orally or  X  group=9  rats.  After 19th week all of rats were then terminated and tumor lesion of colon were processed hystophatologically. The tissues of colon cancer were stained by H&E for evaluate the lymphocytes infiltration and immunohistochemistry monoclonal antibody anti-perforin for perforin expression. Non pairs t-test was used with considered significant if p<0.05.
Results: The mean of lymphocytes infiltration of the group X was 401.89±70.19, it was higher compared to K+ 191.89±50.68 (p=0.000). The mean percentage of perforin expression of group X was 39.00±1.80%, it was higher compared to K+ 23.00± 3.00% (p=0.000). 
Conclusion: The extract of P. niruri L increases immunologic status through mechanism of lymphocytes infiltration and perforin expression elevation of colon cancer in animal mode.

Keywords : Phyllanthus niruri Linn, colon cancer, lymphocytes infiltration, perforin

ABSTRAK
Latar belakang: Penanganan kanker kolon saat ini melibatkan imunoterapi yang bertujuan untuk meningkatkan kualitas hidup dan survival penderitanya. Phyllanthus niruri Linn bekerja sebagai imunomodulator, tetapi potensinya dalam respons imun antitumor belum banyak diungkap. Penelitian bertujuan untuk mengevaluasi perbedaan status imunologis antara tikus coba yang menderita kanker kolon yang tidak diberi dengan yang diberi ekstrak P. niruri L.

Metode: Penelitian ini merupakan randomized posttest-only control group design. Sampel berupa 30 ekor tikus Sprague-Dawley jantan, yang diinduksi 1,2 DMH 30 mg/kgBB subkutan sekali setiap minggu. Pada minggu ke-9, 11 dan 13 masing-masing empat ekor tikus dimatikan untuk melihat perkembangan tumor. Minggu ke-13 pada kolon empat tikus telah tumbuh kanker kolon, sehingga induksi dihentikan dan sisa 18 tikus dirandom alokasi menjadi 2 kelompok. Kelompok kontrol positif (K+) tanpa pemberian P. niruri L (9 tikus) dan kelompok diberi P. niruri L. 13,5 mg/kg per hari melalui sonde (9 tikus). Minggu ke-19 semua tikus diterminasi, lesi tumor pada kolon diproses menjadi sediaan histopatologik, kemudian dipulas dengan H&E untuk memeriksa infiltrasi limfosit dan pulasan imunohistokimia menggunakan antibodi monoklonal anti-perforin untuk melihat ekspresi perforin. Analisis data menggunakan uji t tidak berpasangan dengan derajat signifikansi p<0,05.
Hasil: Rerata jumlah infiltrasi limfosit kelompok X adalah 401,89±70,19 lebih tinggi dibanding kelompok K+ yaitu191,89±50,68 (p=0,000). Persentase rerata ekspresi perforin kelompok X sebesar 39,00±1,80%, lebih tinggi dibandingkan dengan K+ yakni 23,00±3,00% (p=0,000).
Simpulan: Ekstrak P. niruri L meningkatkan status imunologi melalui mekanisme peningkatan infiltrasi limfosit dan ekspresi perforin untuk melawan kanker kolon pada tikus coba




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