SASARAN AKSI EKSTRAK AIR HERBA CIPLUKAN (Physalis angulata L) TERSTANDAR FISALIN PADA JALUR SINTESIS NITRIC OXIDE TIKUS SPRAGUE DAWLEY DIINDUKSI NICOTINAMIDE DAN STREPTOZOTOCIN

YENY SULISTYOWATI, Prof. dr. Sri Kadarsih, S., M.Sc., Ph.D. AIFM

2015 | Disertasi | S3 Ilmu Kedokteran

Hiperglikemia pada penderita DM meningkatkan stres oksidatif pada endotel dan terjadi disfungsi endotel. Disfungsi endotel terkait stres oksidatif akibat hiperglikemi diperbaiki melalui beberapa jalur mekanisme, diantaranya jalur sintesis nitric oxide (NO). Herba ciplukan (Physalis angulata L) salah satu jenis tanaman antidiabetes, karena mengandung fisalin. Pemberian ekstrak air herba ciplukan (P. angulata L) terstandar fisalin sebagai antioksidan terkait ADMA, DDAH2, VEGF, eNOS dan NO pada jalur sintesis NO belum pernah diteliti. Penentuan letak kerja ekstrak air herba ciplukan (P. angulata L) terstandar fisalin dengan pemberian bersama insulin dan atau verapamil perlu dilakukan pada hewan coba diinduksi nicotinamide dan streptozotocin, sehingga diketahui sasaran aksinya terkait ADMA, VEGF, NO, sebelum DDAH2, setelah DDAH2 atau langsung eNOS. Jenis penelitian eksperimen murni, dengan rancangan pre-and post-test only control group design. Subyek penelitian 30 ekor tikus Sprague Dawley diinduksi nicotinamide (110 mg/kgBB) dan streptozotocin (45 mg/kgBB). Pemberian ekstrak air herba ciplukan (P. angulata L) tertandar fisalin dosis 20 mg/kgBB selama 21 hari. Suntikan insulin 9 IU/kgBB dan verapamil 1 mg/kgBB dilakukan pada hari ke 31. Hari ke-10, ke-31 dan ke-32 dilakukan pemeriksaan kadar ADMA, VEGF, dan NO dengan ELISA. Pemeriksaan ekspresi DDAH2 dan eNOS dilakukan pada hari ke-32 dengan imunohistokimia. Uji normalitas dilakukan dengan Saphiro-Wilk, mengetahui perbedaan dengan t test, ANOVA dan regresi. Pemberian ekstrak air herba ciplukan (P. angulata L) terstandar fisalin menghasilkan ekspresi DDAH2, kadar VEGF dan kadar NO lebih tinggi dibandingkan tanpa ekstrak.. Pemberian ekstrak air herba ciplukan (P. angulata L) terstandar fisalin bersama insulin menghasilkan kadar VEGF lebih tinggi dibanding insulin saja, pemberian bersama verapamil menghasilkan kadar VEGF dan ekspresi eNOS lebih tinggi dibanding verapamil saja. Ekstrak air herba ciplukan (P. angulata L) terstandar fisalin bersama insulin tidak menunjukkan hasil signifikan terhadap kadar ADMA, ekspresi DDAH2, kadar VEGF, ekspresi eNOS dan kadar NO dibandingkan ekstrak air herba ciplukan (P. angulata L) terstandar fisalin saja pada tikus hiperglikemia. Ekstrak air herba ciplukan (P. angulata L) terstandar fisalin bersama verapamil menghasilkan ekspresi DDAH2 dan ekspresi eNOS lebih tinggi dibandingkan ekstrak air herba ciplukan (P. angulata L) terstandar fisalin saja pada tikus hiperglikemia. Sasaran aksi ekstrak air herba ciplukan (P. angulata L) terstandar fisalin pada jalur sintesis NO melalui peningkatan kadar VEGF dan kadar NO, sebelum dan sesudah DDAH2 serta langsung eNOS sehingga terjadi sintesis NO. Kata kunci: Fisalin, Insulin, Verapamil, ADMA, DDAH2, VEGF, eNOS, NO.

Hyperglycemia in patients with diabetes increases oxidative stress in endothelial and endothelial dysfunction. Endothelial dysfunction related to oxidative stress due to hyperglycemia is cured through several paths mechanisms, such as synthesis pathway nitric oxide (NO). Ciplukan herb (Physalis angulata L) is a type of antidiabetic plants containing fisalin. Water extract of the ciplukan herbs (P. angulata L) standardized fisalin as an antioxidant linked with ADMA, DDAH2, VEGF, eNOS and NO in NO synthesis pathways have not been investigated. Determining the location of work in water extract of ciplukan herbs (P. angulata L) standardized fisalin by giving insulin and or verapamil should be done in experimental animals induced by nicotinamide and streptozotocin, so it can show the action targets related to ADMA, VEGF, NO, before DDAH2, after DDAH2 or directly eNOS. This research used the true experimental research, the design of pre-and post-test only control group design. There were 30 research subjects of Sprague Dawley rats induced nicotinamide (110 mg/kgBW) and streptozotocin (45 mg/kgBW). Water extract of the herb ciplukan (P. angulata L) standardized with fisalin dose of 20 mg / bw for 21 days. Insulin injections 9 IU/kgBW and verapamil 1 mg/kg BW was conducted on day 31th. On the day 10th, the 31st and 32nd was the examination of ADMA levels, VEGF, and NO by ELISA. The examination of DDAH2 and eNOS expression were performed on day 32 with immunohistochemistry. Normality test was conducted by Shapiro-Wilk, knowing the difference with t test, ANOVA and regression. Giving water extract of the herb ciplukan (P. angulata L) standardized fisalin produce DDAH2 expression, the levels of VEGF and NO levels were higher than without the extract. Giving ciplukan (P. angulata L) standardized fisalin with insulin resulted in higher levels of VEGF than only giving the insulin. By giving the verapamil too, the result in levels of VEGF and eNOS expression was higher than only giving the verapamil. Water extract of herbs ciplukan (P. angulata L) standardized fisalin with insulin did not show significant results on levels of ADMA, DDAH2 expression, levels of VEGF, eNOS expression and NO levels than water extracts of herbs ciplukan (P. angulata L) standardized fisalin only in hyperglycemic rats. Water extract of herbs ciplukan (P. angulata L) standardized fisalin together verapamil resulted in the expression of eNOS expression, DDAH2 and higher than the water extract of the ciplukan herb (P. angulata L) standardized fisalin only in hyperglycemic rats. The action target of ciplukan herb extract water (P. angulata L) standardized fisalin in NO synthesis pathway shows increased levels of VEGF and NO levels, before and after DDAH2 and directly eNOS result NO synthesis. Keywords: Fisalin, Insulin, Verapamil, ADMA, DDAH2, VEGF, eNOS, NO

Kata Kunci : Fisalin, Insulin, Verapamil, ADMA, DDAH2, VEGF, eNOS, NO