hepatocyte cells
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2022 ◽  
Author(s):  
N Govardhana Sagar ◽  
A Rajendra Prasad ◽  
Pushpendra Kumar ◽  
Bharat Bhushan ◽  
P Guru Vishnu ◽  
...  

Abstract RNA interference by short hairpin RNAs (shRNAs) is a widely used post transcriptional silencing mechanism for suppressing expression of the target gene. In the current study, five shRNA molecules each against SCD and SREBP1 genes involved in denovo lipid biosynthesis were designed upon considering parameters such as secondary structures of shRNAs, mRNA target regions, GC content and thermodynamic properties (ΔG overall, ΔG duplex and ΔG break-target), synthesized and cloned in pENTR/U6 entry vector to knockdown the expression of SCD and SREBP1 genes. After transfection of these shRNA constructs into the chicken embryonic hepatocytes, expressions of the target genes were monitored by real time PCR. Significant reduction (P<0.05) in the expression of SCD and SREBP1 genes was observed in hepatocytes. The shRNAs against SCD gene showed the knock down efficiency ranged from 20.4% (shRNA5) to 74.2% (shRNA2). In case of SREBP1 gene, the shRNAs showed knock-down efficiency ranging from 26.8% (shRNA4) to 95.85% (shRNA1). The shRNAs against both the genes introduced in chicken hepatocyte cells did not show any significant impact on expression of immune response genes (IFNA and IFNB) in those cells. These results clearly demonstrated the successful down regulation of the expression of SCD and SREBP1 genes by the shRNA molecules against both the target genes under in vitro condition. It is concluded that the shRNA molecules against SCD and SREBP1 genes showed great potential to silence the expression of these genes under in vitro chicken embryonic hepatocyte cells.


Nanomaterials ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 16
Author(s):  
Irina A. Shurygina ◽  
Galina F. Prozorova ◽  
Irina S. Trukhan ◽  
Svetlana A. Korzhova ◽  
Nataliya N. Dremina ◽  
...  

A new original copper nanocomposite based on poly-N-vinylimidazole was synthesized and characterized by a complex of modern physicochemical and biological methods. The low cytotoxicity of the copper nanocomposite in relation to the cultured hepatocyte cells was found. The possibility to involve the copper from the nanocomposite in the functioning of the copper-dependent enzyme systems was evaluated during the incubation of the hepatocyte culture with this nanocomposite introduced to the nutrient medium. The synthesized new water-soluble copper-containing nanocomposite is promising for biotechnological and biomedical research as a new non-toxic hydrophilic preparation that is allowed to regulate the work of key enzymes involved in energy metabolism and antioxidant protection as well as potentially serving as an additional source of copper.


2021 ◽  
Vol 1 (1) ◽  
pp. 39-49
Author(s):  
Sister Sianturi

ABSTRAK   Penelitian ini dilakukan sebagai lanjutan dari penelitian sebelumnya yaitu uji pemberian air rebusan mie instan pada mencit betina bunting dan diperoleh hasil bahwa terdapat kelainan fetus tetapi pada analisis statistic kelainan tersebut tidak signifikan. Penelitian ini dilakukan untuk mengetahui gambaran histologi organ hepar mencit betina bunting setelah pemberian air rebusan mie instan. Tahapan penelitian ini adalah sebagai berikut: persiapan hewan uji mencit betina bunting yang sudah diberi perlakuan air rebusan mie instan dari 3 merek yang berbeda yaitu P1, P2, dan P3.Masing-masing perlakuan tersebut terdiri atas pemberian air rebusan dengan konsentrasi yang berbeda yaitu 30%, 50%, dan 70% selama 14 hari, kemudian mencit dikorbankan lalu diambil organ hepar,  pembuatan preparat histology, lalu diamati struktur histologinya. Metode pembuatan preparat histologi mengikuti prosedur pewarnaan HE (Haematoxyllin-Eosine). Hasil penelitian menunjukkan adanya perubahan struktur sel hepatosit hepar pada perlakuan yaitu degenerasi midzonal dan nekrosis. Hal ini menunjukkan adanya pengaruh pemberian perlakuan air rebusan mi instan terhadap susunan sel hepar. Penelitian menggunakan jumlah sampel organ yang terbatas sehingga perlu dilakukan penelitian dengan sampel yang lebih banyak dan diperlukan adanya analisis statistik untuk mengetahui tingkat derajat kerusakan sel.   ABSTRACT This study was conducted based on previous research, to test of giving instant noodle boiled water to pregnant female mice and the results showed that there were fetal abnormalities but the statistical analysis of these abnormalities was not significant. This study was conducted to determine the histology of the liver of pregnant female mice after giving instant noodle boiled water. The stages of this research are as follows: preparation of pregnant female mice that have been treated with instant noodle boiled water from 3 different brands, namely P1, P2, and P3. Each treatment consisted of giving boiled water with different concentrations of 30 %, 50%, and 70% for 14 days, then the mice were sacrificed and then the liver was taken, histology preparations were made, then the histological structure was observed. The method of making histology preparations followed the HE (Haematoxyllin-Eosine) staining procedure. The results showed that there were changes in the structure of the liver hepatocyte cells in the treatment, namely midzonal degeneration and necrosis. This shows that there is an effect of giving instant noodle boiled water treatment on the composition of liver cells. The study used a limited number of organ samples, so it is necessary to do research with more samples and statistical analysis is needed to determine the degree of cell damage


2021 ◽  
Vol 10 (3) ◽  
pp. e32010313371
Author(s):  
Walter Félix Franco Neto ◽  
Murilo Tavares Amorim ◽  
Karla Fabiane Lopes de Melo ◽  
Gustavo Moraes Holanda ◽  
Jardel Fábio Lopes Ferreira ◽  
...  

Objective: Evaluate the messenger RNAs (mRNA) PIWI proteins expression during infection with VDEN 4 in human hepatocyte cells. Materials and Methods: VDEN4 strain H778494 (JQ513335) was used, which was stored in Aedes albopictus cell culture (Clone C6 / 36) at the Arbovirology and Hemorrhagic Fevers section of the Evandro Chagas Institute. The techniques of cell cultures, stock (C6/36), inoculation, extraction, viral load quantification, RTqPCR and statistical analyzes were performed at the Viral Biogenesis Laboratory. Results: According to the results obtained, two cells (HepG2 and Huh7.5) demonstrated a higher level of viral replication at 72 hours post infection (hpi). The PIWI 2 target alter its mRNA expression during VDEN4 infection. The PIWI 4 target expression, was observed an altered expression in the infected cells. Thus, it was found that they are in different poles, while the viral load in the first three days showed high expression. The expression of mRNA was low in relation to the normal rate given by the uninfected cells. Conclusion: In our findings, it was observed that PIWI2 and 4 proteins have an inverse relationship to the viral infection process by VDEN4, when there is an increase in viral replication, these two proteins end up having a significant reduction in their expression. Probably, this reduction of expression is involved with the biogenesis process of apoptosis-regulating microRNAs.


2021 ◽  
Vol 10 (1) ◽  
pp. 109
Author(s):  
Oktantia Frenny Anggani ◽  
Aniek Setiya Budiatin ◽  
Laksmi Sulmartiwi ◽  
Muhammad Rahmad Royan

Scaffold, as one of the components for bone tissue engineering, requires formulated biomaterials that are both structurally and compositively similar to bone composition. Among others, chitosan, gelatin and chondroitin sulfate are known as potential candidates for scaffold composites that can be easily obtained from waste-based resources. This study aims to investigate the cytotoxicity of different scaffold composition and concentration regimes derived from waste-based chitosan, gelatine and chondroitin sulfate, in vitro. The composition regimes used were (Chitosan : Gelatin : Chondroitin Sulfate) 50 : 50 : 0 (A); 50 : 40 :10 (B); 50 : 35 : 15 (C); 50 : 30 : 20 (D); 50 : 25 : 25 (E). Meanwhile, the final concentrations of scaffold used were 2000, 1000, 500, 250, 100, 10 and 0,1 mg/ml. The different compositions and concentrations of scaffold was tested against Hepatocellular Carcinoma (Huh7it / Human Hepatocyte It). After 48-hour incubation in the scaffold solution, the percentage of cell viability was evaluated using 3-(4,5-dimethylthiazol-2yl)-5(3-carboxymethoxyphenyl)-2- (4-sulfophenyl)-2H-tetrazolium (MTT) assay. The result shows that there is no difference observed among different scaffold compositions on the cell viability (p > 0.05). However, different concentrations of scaffold show significant differences in cell viability in composition C and E (p < 0.01), suggesting possible dose- dependent effect of scaffold on cell viability. Overall, all the waste-based scaffold compositions show no toxicity against the Hepatocellular Carcinoma cells as exhibited by the cell viability that is above 70%, at least with the concentration up to 2000 mg/ml.


Author(s):  
Kadek Ayu Cintya Adelia

<p><strong>Abstract: </strong>This study examines the effect of the use of gamma radiation in the health sector, one of which is the use of gamma rays in the radiotherapy process. Irradiation into the body can cause the emergence of free radicals in the body which can harm other organs around the irradiation target. This study aims to determine the effect of exposure to gamma radiation on the liver of mice and the hepatoprotective properties of mangosteen peel extract (Garcinia mangostana) in reducing the resulting cell damage. This study was conducted by giving mice mangosteen peel extract with 5 different doses from those given 40 minutes of radiation exposure with a total dose of 268 μSv for 14 days, then the SGPT (serum glutamic pyruvic transaminase) levels were measured and cell damage figures from histopathological preparations. The results showed that before being given mangosteen peel extract, total damage to hepatocyte cells was 47.34%. After being given mangosteen peel extract, total damage to hepatocyte cells was 23.42%. Meanwhile, the SGPT level decreased to 77.7 U/L from the initial level of 128.67 U/L. This proves that mangosteen peel extract has hepatoprotective properties that can reduce cell damage due to exposure to gamma radiation.</p><p class="AbstrakIndonesia"><strong>Abstrak: </strong>Penelitian ini mengkaji pengaruh penggunaan radiasi gamma di bidang kesehatan, salah satunya adalah penggunaan sinar gamma dalam proses radioterapi. Penyerapan energi radiasi ke dalam tubuh dapat menyebabkan munculnya radikal bebas dalam tubuh yang dapat merugikan organ lain di sekitar target penyinaran. Penelitian ini bertujuan untuk mengetahui pengaruh paparan radiasi gamma terhadap hepar mencit dan sifat hepatoprotektif ekstrak kulit manggis<em>(Garcinia mangostana)</em> dalam mengurangi kerusakan sel yang ditimbulkan. Penelitian ini dilakukan dengan memberikan ekstrak kulit manggis 5 dosis varian berbeda ke mencit dan diberi paparan radiasi selama 40 menit dengan dosis total 268 μSv selama 14 hari, kemudian diukur kadar SGPT <em>(serum glutamic pyruvic transaminase)</em> dan jumlah kerusakan sel dari <em>p</em><em>reparat</em>histopatologis. Hasil penelitian menunjukkan bahwa sebelum diberikan ekstrak kulit manggis, kerusakan total sel hepatosit sebesar 47,34%. Setelah diberikan ekstrak kulit manggis, kerusakan total sel hepatosit sebesar 23,42%. Sementara itu, kadar SGPT turun menjadi 77,7 U/L dari kadar awal 128,67 U/L. Hal ini membuktikan bahwa ekstrak kulit manggis memiliki sifat hepatoprotektif yang dapat mengurangi kerusakan sel akibat paparan radiasi gamma.</p>


2020 ◽  
Vol 1 (2) ◽  
pp. 21
Author(s):  
Dede Renovaldi ◽  
Abdul Khalik Adam

The use of drugs is one of the most common causes of liver injury, because the liver is the main organ that metabolizes drugs. Little is currently done if there is a liver injury due to the hepatotoxic side effects of a drug. Herbal plants have active natural compounds that have pharmacological effects so they are widely used as alternative treatments. Sweet basil (Ocimum basilicum) is one of the most cultivated plants in Asia. Studies on the use of Ocimum basilicum in medicine have been carried out, one of which is the hepatoprotector effect. Studies indicate that Ocimum basilicum is rich in high antioxidant content (flavonoids, saponins, tannins, sterols, triterpenes, and rosmaniric acid) capable of providing hepatoprotector effects by helping the regeneration process of hepatocyte cells that are damaged by hepatotoxic agents and significantly decreasing liver damage biomarkers. The purpose of this review is to explain the potential of Ocimum basilicum as a hepatoprotective agent for liver injury associated with drugs. The conclusion of this review is Ocimum basilicum has high potential in its utilization as a hepatoprotector against liver injury mainly related to the consumption of drugs that have hepatotoxic effects.


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