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Curcumin Improves the Regulation of Ovarian Folliculogenesis in Culture Media with Peritoneal Fluid from Infertile Women with Endometriosis Hendy Hendarto1, M Yohannes Ardianta Widyanugraha1, Widjiati2, Soehartono Darmosoekarto1
Indian Journal of Forensic Medicine & Toxicology Vol. 14 No. 2 (2020): Indian Journal of Forensic Medicine & Toxicology
Publisher : Institute of Medico-legal Publications Pvt Ltd

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.37506/ijfmt.v14i2.3154

Background : Peritoneal fluid (PF) from infertile women with endometriosis contains a variety of inflammatory mediators that may interfere with folliculogenesis. The aim of the study was to evaluate effects of curcumin on the regulation of ovarian folliculogenesis by evaluating Growth Differentiation Factor (GDF)-9, Kit Ligand (KitL) and Tumor Necrosis Factor (TNF)? expressions in bovine cumulus oocyte complexes (COC)s cultured with PF from infertile women with endometriosis. Method: COCs were aspirated from antral follicles of bovine ovaries. PF was collected from infertile women with endometriosis undergoing laparoscopy for infertility evaluation. Curcumin, a strong antiinflammatory turmeric, was added in Tissue Culture Media (TCM)199 and PF from infertile women with endometriosis for culture media. Bovine COCs were cultured into 3 groups of media: 1) TCM199, 2) TCM199+PF from infertile women with endometriosis, and 3) TCM199+PF from infertile women with endometriosis+curcumin. GDF-9, KitL and TNF? expressions were examined using immunohistochemistry technique. Results: GDF-9 expression of bovine COCs cultured in PF from infertile women with endometriosis with curcumin addition (2.67±0.98) was found to increase compared to those cultured without curcumin (0.50±0.67) but reduced compared to the control. This result was similar to KitL expression of bovine COCs cultured with curcumin (2.67±1.23), in which it increased compared to those without curcumin (0.33±0.49) (p<0.05). A significant difference in TNF? expression was noted between groups with or without curcumin (p<0.05). Conclusion: In the culture of PF from infertile women with endometriosis, curcumin addition may improve the regulation of ovarian folliculogenesis through the decrease of inflammation factor.

Moringa Leaf (Moringa oleifera Lam) Nanoparticle Supplementation on Zygote Cleavage in Goat Embryo Culture In Vitro Alfina Aisatus Saadah1 , Widjiati2 , Hendy Hendarto3
Indian Journal of Forensic Medicine & Toxicology Vol. 14 No. 3 (2020): Indian Journal of Forensic Medicine & Toxicology
Publisher : Institute of Medico-legal Publications Pvt Ltd

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.37506/ijfmt.v14i3.10669

One of the causes of failure of zygote development is oxidative stress in the culture medium. Where oxidative stress can damage the structure and function of zygotes due to free radicals Reactive Oxygen Species (ROS). Moringa oleifera Lam contains antioxidant polyphenol compounds, flavonoid flavanols namely quercetin which is the most antioxidant content in Moringa leaves which is expected to fight excess ROS. The purpose of this study was to prove that the supplementation of Moringa oleifera Lam leaf nanoparticles to increase zygote cleavage in goat embryo culture in vitro. The research subjects used goat oocytes which were divided into 4 groups consisting of 1 control group and 3 treatment groups. Each group consisted of 31 samples. Control group K1 without Moringa leaf nanoparticle supplementation, treatment group I P1 Moringa leaf nanoparticle supplementation with a dose of 0.5 µM, treatment group II P2 supplementation of Moringa leaf nanoparticles with a dose of 1.0 µM, Treatment group III P3 supplementation with Moringa leaf 2.0 µM. Embryo culture was carried out in a 5% CO2 incubator, temperature 38.5ºC for 48 hours, then observed under an inverted microscope. The results show that there are significant differences with the value of p = 0.041 which means p <0.05. Supplementation of moringa leaf nanoparticles (Moringa oleifera Lam) can significantly increase zygote cleavage at embryo culture stage in vitro.

Nanocurcumin Potential Effect of SOD Enzyme and Caspase-3 Expression in Lead-Acetate Induced Rats Ovarian Granulosa Cells Anis Satus Syarifah; Sri Agus Sudjarwo; Hendy Hendarto; Reny I’tishom; Supriyanto
Indian Journal of Forensic Medicine & Toxicology Vol. 15 No. 2 (2021): Indian Journal of Forensic Medicine & Toxicology
Publisher : Institute of Medico-legal Publications Pvt Ltd

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.37506/ijfmt.v15i2.14650

Aim: This study investigated the potential effect of nanocurcumin on the increase of SOD enzyme expressionand decrease of caspase-3 in lead acetate-induced rats ovarian granulosa cells.Materials and Methods: Forty five female rats were divided into 5 groups, the negative control group (ratsreceiving corn oil, one hour later receiving distilled water), positive control group (rats receiving corn oil,one hour later receiving lead acetate of 40 mg/kg bw) and experimental groups 1, 2 and 3 (rats receivingnanocurcumin 50 mg, 100 mg and 200 mg/kg bw). One hour after administering the nanocurcumin, therats received 40 mg/kg of lead acetate. All groups received oral treatment once a day for 26 days. Onday 27 the rats were sacrificed and the expression of SOD and caspase-3 enzymes were measured usingimmunohistochemical methods.Results: This study found that lead acetate decreased SOD enzyme expression and increased caspase-3. Incontrast, nanocurcumin increased SOD enzyme and decreased caspase-3 expression in lead acetate-inducedrats ovarian granulosa cells.Conclusion: Nanocurcumin has potential as a strong natural antioxidant by affecting the increase of SODand the decrease of caspase-3 cells in lead acetate-induced rats ovarian granulosa cells.

The Relationship of Antral Follicle Count (AFC) with Levels of Estradiol Serum and Mature Ovarian Follicles in Women Undergoing the IVF Program at the Graha Amerta Fertility Clinic of General Hospital of Dr. Soetomo in the Period 2018 Anak Agung Renjani Kanya Kumari; RenyI’tishom; Hendy Hendarto; Sri Ratna Dwiningsih
Indian Journal of Forensic Medicine & Toxicology Vol. 15 No. 3 (2021): Indian Journal of Forensic Medicine & Toxicology
Publisher : Institute of Medico-legal Publications Pvt Ltd

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.37506/ijfmt.v15i3.15608

Background: In the last two decades, one of the critical factors of assisted reproductive technology (ART)has been identified as ovarian capacity. Consequently, ovarian capacity markers have evolved to be part ofdiagnostic testing conducted regularly before IVF. Several predictor factors, such as antral follicle count(AFC), is one of several factors to predict ovarian capacity. These diagnostic procedures and tests have thepurpose of identifying women at high-risk patient who respond poorly toward controlled ovarian stimulationthroughout the IVF cycle. The ovarian response toward ovulation stimulation within this research was assessedby peak estradiol (E2) serum levels and the total number of mature follicles. There is no clear relationshipbetween AFC and levels of estradiol serum and the number of mature ovarian follicles to identify womenwho respond poorly. Methods: This study is an analytical study with a cross-sectional design. Samples weretaken from secondary data of medical records using the total sampling method on women undergoing the IVFprogram at the Graha Amerta fertility clinic of General Hospital of Dr. Soetomo from January 1 to December31 2018 according to the inclusion and exclusion criteria. The AFC variables, estradiol serum levels, and thenumber of mature ovarian follicles in the study were analyzed using Spearman’s correlation. Conclusion:There is a significant relationship between antral follicle count (AFC) with the levels of estradiol serum andthe number of mature follicles in women undergoing the IVF program.

Penetapan Total Flavonoid, Aktivitas Antioksidan dan Karakterisasi Nanopartikel Ekstrak Etanol Daun Kelor (Moringa pterygosperma Gaertn.) Tinta Julianawati; Hendy Hendarto; Widjiati Widjiati
Jurnal Penelitian Kesehatan SUARA FORIKES Vol 11, No 1 (2020): Januari 2020
Publisher : FORIKES

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33846/sf11110

Moringa pterygosperma is one of the plants in Indonesia that is often used in herbal medicine. Moringa leaves contain many phytochemical compounds, one of the compounds is flavonoids which function as antioxidants. Flavonoid compounds in vitro can inhibit lipid peroxidation by breaking the chain of peroxyl radicals. Moringa leaf benefits in the field of herbal medicine are formed with a variety of preparations. One of the preparations is in the form of extracts with the aim of attracting chemical components to natural materials The extraction process has a disadvantage because the level of solubility in water is still low. The nanoparticle technique is a strategy to increase the bioavalbicibility of herbal active compounds. This research was experimental in order to determine the total flavonoid levels contained in Moringa leaf extract. In addition, it was also to find out the antioxidant activity and particle size of the moringa leaf extract nanoparticles. Determination of flavonoid levels using spectrophotometric methods. The antioxidant activity of moringa leaf extract nanoparticles was tested using the DPPH method (2,2-diphenyl-picrylhdrazyl) and to determine the particle size used by using the Particle Size Analyzer (PSA). The results of the study showed total flavonoid levels in moringa leaf extract (1.97 ± 1.07)%. Antioxidant activity of 451.8 ppm with a reduction of 2.8% and particle size in the moringa leaf extract nanoparticles of ± 134.78 nm. Moringa leaf extract nanoparticles include low antioxidants. Keywords: moringa; nanoparticles; antioxidants

Pengaruh Suplementasi Ekstrak Daun Kelor (Moringa pterygosperma Gaertn) terhadap Ekspansi Kumulus Oosit dalam Medium Maturasi In Vitro Nur Anindya Syamsudi; Widjiati Widjiati; Hendy Hendarto
Jurnal Penelitian Kesehatan SUARA FORIKES Vol 11, No 1 (2020): Januari 2020
Publisher : FORIKES

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33846/sf11109

Background: Cumulus cells are mediators providing energy transport, micronutrients, and carrier molecules for oocyte development, and mediate the influence of hormones on the oocyte cumulus complex. Cumulus cells play a role during oocyte growth and maturation which shows the quality of oocyte maturation. The use of basic maturation medium with the addition of various substances and compounds has been extensively studied to study oocyte maturation in vitro. Utilization of drugs from natural ingredients has unique advantages because in general cheap prices, have lower toxicity and side effects, and good therapeutic potential. One of the plants that can be used as a source of natural antioxidants, namely Moringa leaves. Objective: Determine the effect of supplementation Moringa pterygosperm Gaertn extract on the expansion of oocyte cumulus in in vitro maturation medium. Method: This research was true experimental with Post Test Only Control Group Design approach. Divided into 4 groups, namely K: control group, P1: treatment of 10 mg / mL Moringa leaf extract, P2: treatment of 15 mg / mL Moringa leaf extract, and P3: treatment of 20 mg / mL Moringa leaf extract. Furthermore oocytes are matured for 24 hours. Cumulus expansion was evaluated according to the level of expansion achieved where the minimum response (+), medium response (++), and maximum response (+++). Statistical analysis used the One Way Anova test and continued post HoC LSD. Results: There was an influence of supplementation Moringa pterygosperm Gaertn extract on the expansion of moderate response cumulus (++) (p = 0.007) and maximum response (+++) (p = 0.000), but there was no effect on the expansion of cumulus minimal response (+) (0.065 ). Doses of 20 mg / mL are effective for cumulus expansion (+++), but not effective for cumulus expansion (+), (++), and control. Conclusion: Supplementation Moringa pterygosperm Gaertn extract affected the expansion of moderate response (++) and maximum response (+++) oocytes in in vitro maturation medium. Keywords: moringa leaf extract, cumulus expansion, in vitro maturation. ABSTRAK Latar Belakang: Sel kumulus merupakan mediator penyedia transpor energi, mikronutrisi, dan atau molekul pembawa untuk perkembangan oosit, dan menjadi mediasi pengaruh hormon pada kompleks kumulus oosit. Sel kumulus berperan selama pertumbuhan dan maturasi oosit yang menunjukkan kualitas maturasi oosit. Penggunaan medium maturasi dasar dengan penambahan berbagai zat dan senyawa banyak diteliti untuk mempelajari maturasi oosit secara in vitro. Pemanfaatan obat dari bahan alam memiliki kelebihan unik karena pada umumnya harga murah, memiliki toksisitas dan efek samping lebih rendah, serta potensi terapeutik baik. Salah satu tumbuhan yang dapat digunakan sebagai sumber antioksidan alami, yaitu daun kelor. Tujuan: mengetahui pengaruh suplementasi ekstrak daun kelor (Moringa pterygosperma Gaertn) terhadap ekspansi kumulus oosit dalam medium maturasi in vitro. Metode: Jenis penelitian ini adalah eksperimental murni dengan dengan pendekatan Post Test Only Control Group Design. Dibagi menjadi 4 kelompok, yaitu K: kelompok kontrol, P1: perlakuan 10 mg/mL ekstrak daun kelor, P2 : perlakuan 15 mg/mL ekstrak daun kelor, dan P3 : perlakuan 20 mg/mL ekstrak daun kelor. Selanjutnya oosit di maturasi selama 24 jam. Ekspansi kumulus di evaluasi sesuai dengan tingkat ekspansi yang dicapai dimana respon minimal (+), respon sedang (++), dan respon maksimum (+++). Analisa statistik menggunakan uji one way Anova dan dilanjutkan post HoC LSD. Hasil: Terdapat pengaruh suplementasi ekstrak daun kelor terhadap ekspansi kumulus respon sedang (++) (p=0.007) dan respon maksimal (+++) (p=0.000), namun tidak terdapat pengaruh terhadap ekspansi kumulus respon minimal (+) (p=0.065). Kesimpulan : Suplementasi ekstrak daun kelor (Moringa pterygosperma Gaertn) berpengaruh terhadap ekspansi kumulus oosit respon sedang (++) dan respon maksimal (+++) dalam medium maturasi in vitro. Kata kunci: ekstrak daun kelor, ekspansi kumulus, maturasi in vitro.

Jus Tomat Meningkatkan Kepadatan Tulang Tikus Menopause (TOMATO JUICE INCREASE BONE DENSITY OF MENOPAUSE RATS) Hening Laswati; Hendy Hendarto; Dian Irawati; Laba Mahaputra
Jurnal Veteriner Vol 16 No 3 (2015)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (153.666 KB)

Osteoporosis in postmenopausal women happened due to estrogen deficiency which leads imbalancebone-formation and bone-resorption process. Recently, phytoestrogen as an alternative hormon replacementtherapy in postmenopausal women could overcome estrogen deficiency. The objectives of this study was toevaluate the effect of tomato juice administration and combination of physical exercise on bone density inestrogen deficiency condition. Twenty eight postovarectomy female rats (Rattus novergicus) were randomizedinto four experiment groups : the controlled group ( 2 mL aquades administration); the exercised group(swimming for 30 minutes, three times in one week), the tomato juice administration every day (44 mg/200g body weight), and the combination exercise and tomato juice administration. Four weeks after treatmentthe samples were collected from os femure for morphological examination. The intervention groups showeda significant difference in bone density with the control group (p<0.05). Bone density highest on the tomatojuice administration group. There is significant difference bone density between tomato juice administrationgroup and the combination group (p<0.05), but no significant difference between physical exercise groupand the combination group. Tomato juice showed an increasing bone density on estrogen deficiencycondition and the combination group have the same effect with the exercised group.

Kurkumin Menurunkan Ekspresi Tumor Necrosis Factor (TNF)-α Kompleks Oosit-Kumulus Sapi pada Kultur dengan Zalir Peritoneum Penderita Infertil Terkait Endometriosis MY Ardianta W; Hendy Hendarto; Widjiati Widjiati
Majalah Obstetri dan Ginekologi Vol. 23 No. 3 (2015): September - Desember 2015
Publisher : Universitas Airlangga

Tujuan: Mempelajari pengaruh pemberian kurkumin terhadap ekspresi tumor necrosis factor (TNF)-α kompleks oosit-kumulus (KOK) sapi pada kultur zalir peritoneum penderita infertil dengan endometriosis ringan dan berat.Bahan dan Metode: Penelitian ini merupakan penelitian eksperimental murni yang dilaksanakan di Laboratorium Embrio-logi Fakultas Kedokteran Hewan Universitas Airlangga antara April sampai Agustus 2014. Zalir peritoneum diambil dari penderita infertil dengan endometriosis ringan, endometriosis berat, dan non-endometriosis yang menjalani laparoskopi diagnosis di Klinik Fertilitas Graha Amerta RSUD dr. Sutomo Surabaya, RS Bersalin Putri Surabaya, RSIA Kendangsari Surabaya, RS Universitas Airlangga Surabaya. Masing-masing zalir peritoneum endometriosis ringan dan berat dibagi lagi menjadi zalir peritoneum endometriosis dengan kurkumin dan tanpa kurkumin. Zalir peritoneum non endometriosis dikelompok-kan menjadi zalir peritoneum penderita non endometriosis tanpa dan dengan kurkumin. KOK sapi diambil secara consecutive sampling dari aspirasi folikel antral diameter 3–8 mm dari ovarium sapi yang berasal dari Rumah Potong Hewan Surabaya kemudian secara acak dengan randomisasi sederhana dikultur dalam tujuh kelompok. Kelompok 1 ditempatkan dalam tissue culture medium 199 (TCM-199) saja. Kelompok 2, 3 dan 4 TCM-199 ditambah masing-masing 3% zalir peritoneum penderita infertil dengan endometriosis ringan, berat dan non-endometriosis. Kelompok 5, 6, dan 7 media TCM-199 ditambah masing-masing 3% zalir peritoneum penderita infertil dengan endometriosis ringan, berat dan non-endometriosis dan dengan kurkumin 20 µg/ml.Hasil: Uji Kruskal Wallis menunjukkan perbedaan yang bermakna ekspresi TNF-α pada ketujuh kelompok (p<0,0001). Uji Mann-Whitney juga menunjukkan bahwa ekspresi TNF-α KOK sapi pada kultur kelompok endometriosis ringan dengan kurkumin (ER+C) lebih rendah secara bermakna dibandingkan kelompok endometriosis ringan tanpa kurkumin (ER). Ekspresi TNF-α KOK sapi pada kultur kelompok endometriosis berat dengan kurkumin (EB+C) lebih rendah secara bermakna dibandingkan kelompok endometriosis berat tanpa kurkumin (EB). Uji Mann-Whitney menunjukkan ekspresi TNF-α KOK sapi pada kultur kelompok ER lebih tinggi secara bermakna dibandingkan kontrol ataupun kelompok non endometriosis (NE). Ekspresi TNF-α KOK sapi pada kultur kelompok EB lebih tinggi secara bermakna dibandingkan kontrol ataupun kelompok NE. Ekspresi TNF-α KOK sapi pada kultur kelompok EB tidak didapatkan perbedaan yang bermakna dibandingkan dengan kelompok ER.Simpulan: Pemberian kurkumin dapat dipertimbangkan untuk digunakan sebagai terapi tambahan pada penderita infertil terkait endometriosis.

Stres Infertilitas Menghambat Maturasi Oosit dan Hasil Fertilisasi In Vitro Hendy Hendarto
Majalah Obstetri dan Ginekologi Vol. 23 No. 1 (2015): Januari - April 2015
Publisher : Universitas Airlangga

Tujuan: Melakukan evaluasi hubungan antara stres infertilitas terhadap maturasi oosit dan hasil fertilisasi in vitro.Bahan dan Metode: Penelitian ini adalah studi potong lintang analitik observasional. Dilakukan di klinik Fertilitas Graha Amerta RSUD Dr Soetomo Surabaya. Tercatat 30 subyek penelitian perempuan infertil yang mengikuti fertilisasi in vitro dan telah dilakukan evaluasi tingkat stres dengan menggunakan Infertility Reaction Scale. Maturasi oosit dan hasil fertilisasi dicatat. Hubungan antara stres infertilitas, maurasi oosit dan hasil fertilisasi dihitung dengan Anova.Hasil: Semua subyek penelitian yang mengikuti fertilisasi in vitro mengalami stres infetilitas dari kadar ringan sampai berat. Pada kelompok tingkat stres infertilitas berat didapatkan oosit matur lebih sedikit (p=0.01). Pada kelompok tingkat stres infertilitas ringan didapatkan oosit matur lebih banyak (p=0.00). Tingkat stres infertilitas berat akan menghasilkan fertilisasi lebih sedikit (p=0.00)Simpulan: Semakin berat stres infertilitas akan menghambat maturasi oosit dan hasil fertilisasi in vitro.

M1 M2 macrophage expression in menstrual blood flakes of women with endometriosis Yulisa Haslinda; Ashon Sa’adi; Hendy Hendarto; Faroek Hoesin
Majalah Obstetri dan Ginekologi Vol. 24 No. 2 (2016): May - August
Publisher : Universitas Airlangga

Objectives: to measure and prove the increase of panmacrophage, macrophages M1 and M2 expression and decrease of ratio of M1/M2 in menstrual blood flakes of women with endometriosis. Materials and Methods: This study was a cross sectional observational analytic study conducted on 30 subjects with endometriosis and non-endometriosis. Immunohistochemical staining was done on a sample of menstrual blood flakes of subjects study who were taken at the second or third day of menstrual cycles with CD68 and CD163 antibody to measure the expression of panmacrophage and M2 macrophages. Expression of M1 macrophages is the approach of a reduction expression of panmacrophage with M2 macrophages.Results: Expression of M1, M2 and the ratio M1/M2 in the both of groups had a normal distribution then continued by independent t-test with one-tailed α (0.05). Probability was considered statistically significant at p <0.05 with a confidence interval of 95%. Based on the statistical result, Mφ macrophage expression in endometriosis and control group amounted to 3.62 ± 0.50 and 2.80 ± 0.64 (p =0.0005) with non parametric test. The expression of M1 macrophages in endometriosis group and non endometriosis were respectively 1.40 ± 0.35 and 1.33 ± 0.40 (p =0.3005) and the expression of M2 in both of group, respectively of 2.23 ± 0.41 and 1.47 ± 0.36 (p =0.0005). The ratio of M1/M2, the endometriosis group and non endometriosis, respectively of 0.65 ± 0.20 and 0.92 ± 0.24 (p =0.0015).Conclusion: this study were significant increased in the panmacrophage Mφ, M2 macrophages expression on a woman's menstrual blood flakes endometriosis and significant decreased in ratio M1/M2 in the woman's menstrual blood flakes endometriosis.

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