Pengujian Toksisitas Seluler SNEDDS Ekstrak Daun Kangkung (Ipomoea reptans, Poir) Terstandar

ABSTRAK Kangkung darat (Ipomea reptans Poir) adalah salah satu tanaman di Indonesia yang secara empiris telah dimanfaatkan oleh masyarakat sebagai obat untuk terapi Diabetes Melitus (DM). Riset sebelumnya telah dilakukan untuk memperoleh data ilmiah dari pemanfaatan kangkung darat sebagai terapi DM, dan pengembangannya dalam bentuk sediaan Self Nano Emulsifying Drug Delivery System (SNEDDS). Penelitian ini bertujuan untuk mengetahui sitotoksisitas sediaan SNEDDS ekstrak daun kangkung terstandar dengan menggunakan metode MTT (Methyl-Thiazolyl-Tetrazolium). Pengujian MTT dilakukan dengan cara well plate diplotkan terlebih dahulu untuk sampel uji 70 sumuran (well), kontrol sel 6 sumuran, dan kontrol media 6 sumuran. Terakhir tiap sumuran ditambahkan stopper SDS sebanyak 100µl. Plate dibungkus alumunium foil dan diinkubasi semalaman pada suhu ruang. Hari berikutnya dilakukan pembacaan absorbansi sampel, kontrol media, dan kontrol sel menggunakan ELISA reader. Uji sitotoksik SNEEDS kangkung pada sel vero diperoleh hasil bahwa SNEEDS kangkung tidak menyebabkan kematian pada sel uji. Pada pengujian sel dilakukan pemaparan sampel excipient dari SNEEDS kangkung sebagai pembanding, dengan hasil menyebabkan kematian <50% sel uji pada dua kadar tertinggi dari sampel excipient. Namun pada hasil uji SNEEDS kangkung diperoleh data bahwa toksisitas excipient pada kadar tersebut tidak mempengaruhi timbulnya toksisitas pada sampel SNEEDS kangkung. Sehingga berdasarkan uji in vitro, SNEEDS kangkung tidak toksik terhadap sel vero. Kata kunci: Ipomoea reptans, sitotoksisitas, SNEDDS ABSTRACT Kangkong (Ipomea reptans Poir) is a type of Indonesian plant empirically utilized by many people to treat diabetes mellitus (DM). A study has been conducted to obtain scientific data from the use of kangkong for DM treatment as well as to develop kangkong in SNEDDS preparation. This current study aimed to examine the cytotoxicity of kangkong leaf standardized extract in SNEDDS preparation through the MTT (Methyl-Thiazolyl-Tetrazolium) method. MTT assay was performed by initially plotting well plates for test sample (70 wells), control cell (6 wells), and control medium (6 wells). As much as 100µl SDS stopper was added into each well, and then plates were wrapped in aluminum foil and incubated all night at ambient temperature. On the following day, the absorbance of test sample, control medium, and control cell was identified using ELISA reader. The cytotoxicity test of kangkong SNEDDS on Vero cell lines showed that kangkong SNEDDS did not cause cell death. The cell was tested through exposure of excipient sample from kangkong SNEDDS as a comparison, resulting in <50% cell death by the two highest concentrations of excipient sample. However, the test result of kangkong SNEDDS indicated that excipient toxicity at such concentrations did not affect kangkong SNEDDS. Therefore, based on in vitro test, kangkong SNEDDS is not toxic against Vero cell lines. Keywords: Ipomoea reptans, cytotoxicity, SNEDDS

Immunological and molecular study of Chlamydia trachomatis as causative agent of abortion in Al-Muthanna province

Chlamydiasis during pregnancy should be considered a significant risk factor for adverse pregnancy outcomes in humans. 120 women who had a single or repeated abortion were selected for this study, and they were referred with a physician report for TORCH tests to determine the final diagnosis of pregnancy loss. The control were (40) healthy pregnant women with a history of a normal pregnancy. The innate immunity in abortive women was higher than normal pregnancies, that was estimated by Nitroblue Tetrazolium test done to estimate the phagocytic activity, there was a significant increase (P=0.009) in phagocytic activity in the leukocytes of abortive women which was (22%) higher than that in control. Also, the cellular immune response was higher in abortive women than that in control. Methyl Thiazolyl Tetrazolium assay was performed to estimate lymphocyte transformation index of peripheral blood leukocytes in abortive women. The results of Methyl Thiazolyl Tetrazolium assay showed a significant increase (P=0.001) in the lymphocyte transformation index in the lymphocytes of abortive women which was (27%) higher than control. The serological detection by ELISA showed that anti-C. trachomatis IgG was (14.2%), and the molecular detection by Quantitive Real Time-Polymerase Chain Reaction showed positive results (17.5%) of total abortive women. The present study demonstrated a high level of relationship between C. trachomatis and abortion among women in the study samples. The age group (20-25years) was the most susceptible to chlamydial infection and the infection was higher in recurrent miscarriages than in single miscarriage.

Proteasome Inhibitor MG132 Enhances Sensitivity to Cisplatin on Ovarian Carcinoma Cells In Vitro and In Vivo

BackgroundPlatinum-based combination chemotherapy after surgery is considered a standard treatment; therefore, any recent drug development should be new, effective, and low toxic, and should have a synergistic effect with platinum. This study aimed to observe the growth of SKOV3 cells after treatment with cisplatin by combining with carbobenzoxy-L-leucyl-L-leucyl-L-leucinal (MG132) and to investigate the effect of the relationship between MG132 and cisplatin combination.Materials and MethodsCell growth was detected by methyl thiazolyl tetrazolium assay after treatment with MG132 at 0.5, 1.5, 2.5, 3.5, and 5.0 μg/mL concentrations for 24, 48, and 72 hours; with cisplatin at 1.0, 2.0, 3.0, 4.0, and 5.0 μg/mL concentrations; and with combination with MG132 at 1.5 μg/mL for 24 hours. The apoptotic rates of cells were detected by a flow cytometer after cisplatin treatment at 1.0, 2.0, 3.0, and 4.0 μg/mL concentrations and that combined with MG132 at 1.5 μg/mL concentration for 12, 24, and 36 hours. A total of 20 BALB/c (nu/nu) female nude mice (age, 4–6 weeks; body weight, 17–19 g) were divided into 4 groups: control, MG132, cisplatin, and combination groups. The expression of Caspase3 and Beclin1 was detected by Western blot analysis and reverse transcription-polymerase chain reaction after treatment with 3.0 μg/mL of the cisplatin group and combined treatment with 1.5 μg/mL of MG132 group for 24 hours, respectively.ResultsMethyl thiazolyl tetrazolium assay demonstrated the inhibitory rates, and the flow cytometery showed that the apoptotic rates in the combination group were higher than those in the cisplatin group (P < 0.01). Western blot analysis and reverse transcription-polymerase chain reaction detected that Caspase3 and Beclin1 at a relative quantity in the combination group were higher than those in the cisplatin group (P < 0.05).ConclusionsMG132 has a synergistic antitumor effect by combining with cisplatin, and it is expected to be an effective antitumor drug for platinum-resistant refractory ovarian cancer.