The Study of Yin-Chen-Hao-Tang Preventing and Treating Alcoholic Fatty Liver Disease through PPAR Signaling Pathway Based on Network Pharmacology and RNA-Seq Transcriptomics

Background. Alcoholic fatty liver disease (AFLD) is the first stage of the alcoholic liver disease course. Yin-Chen-Hao-Tang (YCHT) has a good clinical effect on the treatment of AFLD, but its molecular mechanism has not been elucidated. In this study, we tried to explore the molecular mechanism of YCHT in improving hepatocyte steatosis in AFLD mice through network pharmacology and RNA sequencing (RNA-Seq) transcriptomics. Methods. Network pharmacological methods were used to analyze the potential therapeutic signaling pathways and targets of YCHT on AFLD. Then, the AFLD mice model was induced and YCHT was administered concurrently. Liver injury was measured by serum alanine aminotransferase (ALT) activity and liver tissue H&E staining, and liver steatosis was determined by serum triglyceride (TG) level and liver tissue Oil Red staining. The molecular mechanism of YCHT on prevention and treatment of mice AFLD was investigated according to the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of the differential expression genes data obtained by liver tissue RNA-Seq. Finally, ethanol-induced AFLD AML12 hepatocyte model was established, YCHT with or without PPARα agonist pemafibrate or PPARγ inhibitor GW9662 was administered, Nile Red fluorescent staining was used to evaluate steatosis levels in AML12 hepatocytes, and qRT-PCR was used to detect PPARα and PPARγ gene expression. Results. The results of network pharmacology analysis showed that YCHT may exert its pharmacological effect on AFLD through 312 potential targets which are involved in many signaling pathways including the PPAR signaling pathway. AFLD mice experiments results showed that YCHT markedly decreased mice serum ALT activity and serum TG levels. YCHT also significantly improved alcohol-induced hepatic injury and steatosis in mice livers. Furthermore, KEGG pathway enrichment results of RNA-Seq showed that the PPAR signaling pathway should be the most relevant pathway of YCHT in the prevention and treatment of AFLD. AFLD hepatocyte model experiment results showed that YCHT could remarkably reduce hepatocyte steatosis through reducing PPARγ expression and increasing PPARα expression. Conclusions. Our study discovered that PPARγ and PPARα are the key targets and the PPAR signaling pathway is the main signaling pathway for YCHT to prevent and treat AFLD.

The Effect of Lactation Stage on the Hematological and Serum-Related Biochemical Parameters of the Travnik Pramenka Ewes

The aim of the present research was to determine the hematological and biochemical parameters of the Travnik pramenka ewes during lactation. Thirty‐six ewes in average age of 3-5 years were included in the research. They were monitored in three stages of lactation: early, medium, and late stage. The WBC and RBC, HGB concentration, HCT and MCH values increased significantly as lactation progressed, while significant decrease of MCV and MCHC was determined in blood. HCT was slightly higher in the early compared to late stage, and the MCH in the medium compared to the late stage of lactation. A significant increase in the concentrations of Ca, P, Mg, Na and Cl, as well as a decrease of the Fe concentration was determined. As lactation progressed, the concentrations of urea, total proteins, albumin, globulin and BHB increased, as well as ALT activity. Contrary, concentrations of glucose and NEFA decreased, as well as GGT activity. Obtained results may help in monitoring the health and nutritional status of Travnik ewes. The effect of lactation stage needs to be included in the model when preparing the reference values for the blood parameters of Travnik pramenka ewes, particularly for RBC profiles, energy, and protein metabolites as well as liver enzymes.

The effect of the application of diets with varied proportions of arginine and lysine on biochemical and antioxidant status in turkeys

The aim of the study was to determine the effect of two proportions of arginine (95% and 105%) relative to lysine (Lys), where Lys content in the diet is in accordance with NRC (1994) recommendations or 10% higher, on the metabolism, antioxidant status, and growth performance of turkeys. The experiment had a 2x2 factorial design with two levels of Lys and Arg. The diets with a low level of Lys were according to the NRC (1994) requirements. In the diets with a high level of Lys, the content of Lys was increased by 10% relative to the low level Lys. The two Arg levels in the experimental diets were determined so as to provide 95% and 105% Arg relative to the content of dietary Lys. An increase in the amount of Lys in the diet of turkeys by 10% relative to NRC nutritional recommendations (1994) was not shown to improve growth performance, but had beneficial effects on the metabolism and antioxidant status of the birds, as evidenced by the improvement of hepatic indices (reduction of AST and ALT activity at 9th week of life) and renal indices (reduction of UREA at 9th week of life and reduction of TP and increase level of ALB levels at 16th week of life), as well as an increase in the level of glutathione with strong antioxidant properties at 16th week of life. In comparison to the lower level of Arg in the diet, an increase in the amount of this amino acid to 105% Lys did not improve growth performance, metabolism, or antioxidant status. An Arg level of 95% Lys can be used in a diet for turkeys containing 10% more Lys than the level recommended by the NRC (1994).

COMPLEX FEED ADDITIVE TO NORMALIZE METABOLIC PROCESSES OF COWS

Изучен способ нормализации обменных процессов и повышения продуктивности молочных коров при использовании комплекса биологически активных веществ. Эксперимент проведен на коровах голштинской породы (n=12) с продуктивностью за предыдущую лактацию 9000 кг молока. За 20 дней до отела и в течение 65 дней после коровы опытной группы получали комплексную кормовую добавку (ККД), включающую минерал шунгит, холин в «защищенной» форме, пробиотик «Целлобактерин+» и жмых льняной, в количестве 200 г на голову в сутки в смеси с концентратами. Для изучения обменных процессов в организме коров (n=5) в конце опыта отобраны пробы крови и определены основные биохимические показатели. Для оценки продуктивных качеств проводили контрольные дойки. Отмечено повышение в крови коров опытной группы концентрации общего белка на 2,8%, глюкозы — на 26,8% (P≤0,01), фосфолипидов — на 15,2% (P≤0,05), снижение мочевины на 23,6% (P≤0,05), билирубина — на 23,8% (P≤0,01), активности АЛТ на 13,11 (P≤0,05), АСТ — на 14,6% (P≤0,05). Использование ККД позволило снизить и привести в норму активность ферментов: лактатдегидрогеназы (ЛДГ), гаммаглутаминтрансферазы (ГГТ), креатинкиназы. Повышение интенсивности обменных процессов и нормализация показателей, характеризующих работу печени с использованием в питании комплекса биологически активных веществ ферментно-пробиотического, антиоксидантного и липотропно-гепатопротекторного действия, привело к повышению молочной продуктивности. A method of normalizing metabolic processes and increasing the productivity of dairy cows using a complex of biologically active substances has been studied. The experiment was carried out on Holstein cows (n = 12) with a productivity for the previous lactation of 9000 kg of milk. 20 days before calving and within 65 days after the cows of the experimental group received a complex feed supplement (CFS), including the mineral shungite, choline in a "protected" form, the probiotic "Cellobacterin +" and flaxseed cake, in an amount of 200 g per head per day mixed with concentrates. To study metabolic processes in the body of cows (n = 5), at the end of the experiment, blood samples were taken and the main biochemical parameters were determined. To assess the productive qualities, control milking was carried out. An increase in the blood of cows of the experimental group was noted in the concentration of total protein by 2.8%, glucose - by 26.8% (P≤0.01), phospholipids — by 15.2% (P≤0.05), a decrease in urea by 23,6% (P≤0.05), bilirubin - by 23.8% (P≤0.01), ALT activity by 13.11 (P≤0.05), AST — by 14.6% (P≤ 0.05). The use of CFS made it possible to reduce and normalize the activity of enzymes: lactate dehydrogenase (LDH), gamma glutamine transferase (GGT), creatine kinase. An increase in the intensity of metabolic processes and the normalization of indicators characterizing the work of the liver with the use of a complex of biologically active substances of enzymatic-probiotic, antioxidant and lipotropic-hepatoprotective action in the diet led to an increase in milk productivity.

PROTECTIVE EFFECT OF CURCUMIN RIMPANG CURCUMA LONGA LINN. EXTRACT ON CHLORPYRIFOS POISONING IN WISTAR RATS

This study used an experimental method in the laboratory with a randomized design with five treatments and six repetitions. The parameters examined were cholinesterase (ChE), Aspartate Aminotransferase (AST), and serum Alanine Aminotransferase (ALT) of blood in the first and second treatment stages. Data were analyzed using factorial ANOVA test and LSD multiple comparison test with 95% confidence level with R software version 3.6.2. Wistar rats were given chlorpyrifos (6.75, 13.5, 27, 54) mg/kg BW/day orally once a day for 28 consecutive days, then further intervention with curcumin (27, 54, 108 and 216) mg/kg BW/day once a day for 14 consecutive days. A significant increase in ChE activity and a significant decrease in AST and ALT activity. This study shows that curcumin from Curcuma longa Linn rhizome extract provides a protective effect against chlorpyrifos poisoning in Wistar rats.

Pyrrolizidine Alkaloid-Induced Hepatotoxicity Associated with the Formation of Reactive Metabolite-Derived Pyrrole–Protein Adducts

Pyrrolizidine alkaloids (PAs) with 1,2-unsaturated necine base are hepatotoxic phytotoxins. Acute PA intoxication is initiated by the formation of adducts between PA-derived reactive pyrrolic metabolites with cellular proteins. The present study aimed to investigate the correlation between the formation of hepatic pyrrole–protein adducts and occurrence of PA-induced liver injury (PA-ILI), and to further explore the use of such adducts for rapidly screening the hepatotoxic potency of natural products which contain PAs. Aqueous extracts of Crotalaria sessiliflora (containing one PA: monocrotaline) and Gynura japonica (containing two PAs: senecionine and seneciphylline) were orally administered to rats at different doses for 24 h to investigate PA-ILI. Serum alanine aminotransferase (ALT) activity, hepatic glutathione (GSH) level, and liver histological changes of the treated rats were evaluated to assess the severity of PA-ILI. The levels of pyrrole–protein adducts formed in the rats’ livers were determined by a well-established spectrophotometric method. The biological and histological results showed a dose-dependent hepatotoxicity with significantly different toxic severity among groups of rats treated with herbal extracts containing different PAs. Both serum ALT activity and the amount of hepatic pyrrole–protein adducts increased in a dose-dependent manner. Moreover, the elevation of ALT activity correlated well with the formation of hepatic pyrrole–protein adducts, regardless of the structures of different PAs. The findings revealed that the formation of hepatic pyrrole–protein adducts—which directly correlated with the elevation of serum ALT activity—was a common insult leading to PA-ILI, suggesting a potential for using pyrrole–protein adducts to screen hepatotoxicity and rank PA-containing natural products, which generally contain multiple PAs with different structures.

Inhibition Effect of Gold (III) Theophylline Nano-complex on ALT Enzyme Activity in Human Serumof Iraqi Patients with Liver Disease

Introduction: The ultrasonic sonication approach was used to create an Au(III) nano complex with theophylline. Methods: To explore and suggest the structure of the nano complex, UV-Vis spectroscopy, Fourier transform infrared (FT-IR), and carbon hydrogen and nitrogen (CHN) elemental studies were used. The FE-SEM was used to prove the nanoscale of the prepared complex and was to be less than 20 nm. The effect of the gold nano complex (Au (THP)2(Cl)2) on alanine transaminase (ALT) activity in the serum of chronic liver disease patients was investigated. Results: Compared to the control group, the patients with chronic liver disease with and without nano complex had a significant rise in serum levels of ALT activity (P < 0.001). Furthermore, in individuals with chronic liver disease who received nano complex, the blood levels of ALT activity were significantly lower than those who did not receive nano complex. The reason is that the Au nano complex aggressively interacts with carboxylic groups of important enzymes and inactivates them; further, the Au nano complex had an inhibitory effect on serum ALT activity.

Pengaruh Pemberian Curcumin Terhadap Perbaikan Fungsi Hepar Tikus Putih (Rattus Novergicus) yang diinduksi Parasetamol Dosis Tinggi

Paracetamol has analgesic properties comparable to NSAIDs, but paracetamol have minimal side effects. Paracetamol is metabolized via sulfation and glucuronidation conjugation which is then excreted in the urine. A small part of the paracetamol has been changed to NAPQI. NAPQI will be detoxified by gluthathione. In high doses, there in an increase in NAPQI and a decrease in glutathione levels that results in oxidative stress and liver cell necrosis. Curcumin is often used as a traditional medicine to treat liver disease where it contains phenolic groups capable to scavenge free radicals. Curcumin extract can improve cellular responses to oxidative stress such as increasing the expression of Nrf2, SOD, and gluthathione. The purpose of this research was to know the effect of curcumin on the improvement of liver function in white rats (Rattus novergicus) induced by high dose paracetamol. The design of this research was a descriptive research using literature studies from at least 15 international journals indexed by Scimago or national journals indexed by Sinta published in 2015-2020. Based on the journals used in this research, giving curcumin at a dose of 200 mg/kg BW/day for 2 weeks was effective in significantly increasing gluthathione levels in rats receiving high dose paracetamol. Giving curcumin at a dose of 100 mg/kg BW/day for 7 days can reduce AST and ALT activity in rats receiving high dose paracetamol, but the dose of curcumin that was more effective in reducing AST and ALT activity was 200 mg/kg BW/day for 2 weeks. This is because of curcumin which functions as a hepatoprotector that bind directly to the toxic metabolite of paracetamol, thereby reducing the use of glutathione and quench free radicals, so that oxidative stress in the liver decreased and gluthathione levels increased, AST and ALT activity decreased.

Inhibition of Farnesoid X Receptor Rescues Fat Depression Induced by Dietary Berberine in Grass Carp, Ctenopharyngodon Idella

Berberine (BBR) depresses lipid accumulation in fish, but the mechanism remains unknown. In this study, we hypothesize FXR signaling participates in this physiological process of grass carp. Three diets, namely the control, BBR (1.0 g/kg), and BBR + Gly-β-MCA (an FXR inhibitor) were formulated to feed juvenile grass carp (9.90 ± 0.07) for 8 weeks. Fish fed BBR presented significantly lower IPF index, hepatic TG and TC contents, as well as whole body lipid levels, whereas these were rescued by Gly-β-MCA. Serum TG and HDL-c contents were significantly decreased in fish fed BBR compared to those in the control. The serum ALT activity, combined with the TG, TC, HDL-c, and LDL-c concentrations were all significantly increased in fish fed BBR + Gly-β-MCA than those fed BBR. Dietary BBR significantly increased the mRNA and protein expression of FXR, decreased the mRNA level of FGF19 in the intestine, whereas these were reversed by Gly-β-MCA. In the hepatopancreas, the inhibitor recovered the suppression of the CYP7A1, CYP8B1, and CYP27A1 expression induced by dietary BBR. Fish fed BBR showed significantly lower mRNA expression of SREBP-1c and FAS, whereas these two genes were all up-regulated in response to inhibitor. Dietary BBR increased the gene expression of PPARα, ATGL, CPT-1, which were all abolished by dietary Gly-β-MCA treatment. Fish fed BBR and BBR + Gly-β-MCA showed significantly lower total OTUs, ACE index, chao 1 index, and simpson index of the gut microbiota. Overall, our results demonstrate that inhibition of FXR leads to the rescue of lipid suppression induced by dietary BBR in grass carp.