Study on the mechanism of Salvia miltiorrhiza polysaccharides in relieving liver injury of broilers induced by florfenicol.

In order to explore the transcriptomics and proteomics targets and pathways of Salvia miltiorrhiza polysaccharides (SMPs) alleviating florfenicol (FFC)-induced liver injury in broilers, 60 1-day-old broilers were randomly divided into 3 groups: control group ( GP1) was fed tap water, FFC model (GP2) was given tap water containing FFC 0.15 g/L, and SMPs treatment group (GP3) was given tap water containing FFC 0.15 g/L and SMPs 5 g/L. Starting from 1 day of age, the drug was administered continuously for 5 days. On the 6th day, blood was collected from the heart and the liver was taken. Then 3 chickens were randomly taken from each group, and their liver tissues were aseptically removed and placed in an enzyme-free tube. Using high-throughput mRNA sequencing and TMT-labeled quantitative proteomics technology, the transcriptome and proteome of the three groups of broiler liver were analyzed, respectively. The results of the study showed that the liver tissue morphology of the chicks in the GP1 and GP3 groups was complete and there were no obvious necrotic cells in the liver cells. The liver tissue cells in the GP2 group showed obvious damage, the intercellular space increased, and the liver cells showed extensive vacuolation and steatosis. Compared with the GP1 group, the daily gain of chicks in the GP2 group was significantly reduced (P < 0.0 5 or P < 0.01). Compared with the GP2 group, the GP3 group significantly increased the daily gain of chicks (P <0.0 5 or P <0.01). Compared with the GP1 group, the serum levels of ALT, AST, liver LPO, ROS, and IL-6 in the GP2 group were significantly increased (P < 0.0 5 or P < 0.01), and the contents of T-AOC, GSH-PX, IL-4, and IL-10 in the liver were significantly decreased (P < 0.0 5 or P < 0.01). After SMPs treatment, the serum levels of ALT, AST, liver LPO, ROS, and IL-6 were significantly reduced (P < 0.0 5 or P < 0.01), and the contents of T-AOC, GSH-PX, IL-4, and IL-10 in the liver were significantly increased (P < 0.0 5 or P < 0.01). There were 380 mRNA and 178 protein differentially expressed between GP2 group and GP3 group. Part of DEGs was randomly selected for QPCR verification, and the expression results of randomly selected FABP1, SLC16A1, GPT2, AACS, and other genes were verified by QPCR to be consistent with the sequencing results, which demonstrated the accuracy of transcriptation-associated proteomics sequencing. The results showed that SMPs could alleviate the oxidative stress and inflammatory damage caused by FFC in the liver of chicken and restore the normal function of the liver. SMPs may alleviate the liver damage caused by FFC by regulating the drug metabolism-cytochrome P450, PPAR signaling pathway, MAPK signaling pathway, glutathione metabolism, and other pathways.

Proteome and transcriptome analysis revealed florfenicol via affected drug metabolism and lipid metabolism induce liver injury of broilers.

In order to explore the mechanism of liver injury induced by florfenicol (FFC) in broilers. Sixty broilers were randomly divided into 2 groups: control group: normal drinking water and feed were given every d; FFC group: tap water containing FFC (0.15g/L) was given every d and feed was taken freely; each group was given 5 dd of continuous medication and feed was taken freely. The results showed that compared with the control group, FFC could significantly inhibit the weight gain of broilers (P < 0.05), and significantly inhibit the expression of CYP1A1 and CYP2H1 in liver tissue (P < 0.05). It was found that the expression of genes related to the effect of cytochrome P450 on the metabolism of exogenous substances, the peroxisome proliferators-activated receptors signal pathway, peroxisome pathway and glutathione metabolic pathway in the liver of broilers. The results of qPCR of UDP glucuronosyltransferase family 2A1 (UGT2A1), glutathione S-transferase-like 2 (GSTAL2), hematopoietic prostaglandin D synthase (HPGDS), glutathione S-transferase theta 1(GSTT1), isocitrate dehydrogenase (NADP(+)) 1 (IDH1), acyl-CoA oxidase 2 (ACOX2), fatty acid binding protein 1 (FABP1), adenylosuccinate lyase (ADSL), and phosphoribosyl aminoim idazolesuccino carboxamide synthase (PAICS) genes which were randomly selected from the most significant genes were consistent with those of RNA-seq. The results showed that FFC can affect the drug metabolism and lipid synthesis in the liver of broiler, thus impairing the normal function of liver and the growth and development of broiler.

High-throughput sequencing-based analysis of the intestinal microbiota of broiler chickens fed with compound small peptides of Chinese medicine.

The objective of this study was to determine the effects of compound small peptides of Chinese medicine (CSPCM) on the intestinal microbiota of broilers. A total of thirty-six 1-day-old Arbor Acres broilers were assigned to 6 dietary treatments that include 250, 500, and 750 g/T of CSPCM in feed, 100 g/T of Bacillus subtilis and Clostridium butyricum in feed, and 100 g/T of 50,000 IU xylanase in feed. Each treatment had 2 replicates with 2 cages (3 birds per cage). The jejunal digesta samples were collected from chickens at 42 d. Operational taxonomic unit analysis showed that adding CSPCM at a concentration of 750 g/T of feed can increase the number of operational taxonomic unit samples than other groups. Compared with the control group, adding 250 g/T of CSPCM of feed can improve content of Lactobacillus, Cupriavidus, Ochrobactrum, Candidatus_Arthromitus, Acinetobacter, and Sphingomonas. Adding 500 g/T of CSPCM in feed resulted in varying degrees of improvement in Candidatus_Arthromitus, Acinetobacter, and Sphingomonas. Adding 750 g/T of CSPCM in feed can increase the content of Lactobacillus and Candidatus_Arthromitus. In PICRUSt function prediction analysis, CSPCM acts on the body by creating an environment suitable for the growth of beneficial bacteria. Adding 250 g/T of CSPCM in feed can improve amino acid metabolism, endocrine system function, membrane transport, and cell mobility function. Adding 500 g/T of CSPCM in feed can improve replication and repair and membrane transport function. Adding 750 g/T of CSPCM in feed can increase carbohydrate metabolism, replication and repair, and membrane transport function. Adding B. subtilis and C. butyricum in feed increased replication and repair and membrane transport function. Adding xylanase in feed increased membrane transport function. In conclusion, this study demonstrated that dietary supplementation of CSPCM to broiler diets increased beneficial flora content, metabolism of carbohydrates, amino acid metabolism, the deposition of proteins, renewal of bacteria, and maintenance of vigorous vitality. Among the 3 additive quantities of 250 g/t, 500 g/t, and 750 g/t of CSPCM in feed, 250 g/t of CSPCM improved parameters that are necessary for improved growth and production.

Florfenicol induces renal toxicity in chicks by promoting oxidative stress and apoptosis.

To explore the mechanism of renal toxicity induced by florfenicol (FFC), 120 chicks were randomly divided into 6 groups, 20 in each group. Except for the control group, different doses of FFC (0.15 g/L, 0.3 g/L, 0.6 g/L, 1.2 g/L, and 1.8 g/L) were added to drinking water in the other 5 groups. Five days later, blood was collected from the vein under the wing, and the complete kidneys were obtained as soon as possible, then tested the experimental indicators. The results showed that compared with control group, all doses of FFC significantly reduced the average weight gain of chicks (P < 0.05 or P < 0.01). Except for the 0.15 g/L FFC group, kidney index of chicks in the other doses of FFC groups were significantly increased (P < 0.05 or P < 0.01). The kidney tissues in all FFC groups showed obvious damage, deformities, cell atrophy, and cell gap enlargement. In addition, all doses of FFC significantly increased the contents of uric acid (UA), blood urea nitrogen (BUN), creatinine (CRE) in serum, and malondialdehyde (MDA) in renal tissue (P < 0.05 or P < 0.01), but significantly reduced the levels of glutathione (GSH), superoxide dismutase (SOD), and catalase (CAT) in renal tissue (P < 0.05 or P < 0.01). FFC significantly inhibited the mRNA and protein expression levels of nuclear factor-erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and nicotinamide adenine dinucleotide phosphate: quinone oxidoreductase-1 (NQO-1), and increased the mRNA and protein expression levels of p53, Caspase-3, and Caspase-6 (P < 0.05 or P < 0.01). The apoptotic rate of renal cells in all doses of FFC groups increased significantly (P < 0.05 or P < 0.01). It was concluded that FFC had a certain degree of nephrotoxicity, and with the increase of FFC concentration, the kidney injury of chicks became more and more serious. FFC promoted oxidative stress response in kidney of chicks by inhibiting the expression of related factors in Nrf2-ARE pathway. Moreover, the expression of pro-apoptotic factors was upregulated to improve the apoptosis rate of renal cells, which resulted in excessive apoptosis of renal cells and seriously affected the kidney function of chicks.

Bisphenol A in utero exposure induces ovary dysfunction in mice offspring and the ameliorating effects of Cuscuta chinensis flavonoids.

To study the alleviating effects of flavonoids from Cuscuta chinensis (CCFs) on ovary injury in female offspring of pregnant mice exposed to BPA, five groups (n = 20) of pregnant mice were intragastrically administrated with BPA (5 mg/kg/day) and CCFs (20 mg/kg/day, 30 mg/kg/day, 40 mg/kg/day) at pregnancy days 1-18. The ovaries and serum of F1 female mice were collected at postnatal day (PND) 21 and PND 56 for the detection of related indicators. The ovarian and testicular histomorphologies were observed with hematoxylin-eosin staining (H&E). The levels of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) and the contents of estradiol (E2), progesterone (P4), and testosterone (T) in serum were detected by radioimmunoassay. The contents of ovarian and testicular estrogen receptor alpha (ERα) and estrogen receptor beta (ERß) were detected by enzyme-linked immunosorbent assay (ELISA). The expression of caspase-7, caspase-9, bcl-2, and bax in ovaries and testes of offspring mice were detected by Western blot, and apoptosis in ovaries and testes was detected by TUNEL. The mRNA relative transcription levels of ERα, progesterone receptor (PgR), DNA methyltransferase1 (Dnmt1), DNA methyltransferase3A (Dnmt3A), and DNA methyltransferase3B (Dnmt3B) were detected by real-time quantitative PCR (RT-qPCR). The ovary of female offspring with PND 56 was treated with bisulfite sequence PCR (BSP). Our results showed that, compared with the BPA group, 40 mg/kg CCFs significantly reduced the ovarian index of F1 females and the ovarian cytoapoptosis (P < 0.01). CCFs also can alleviate the injure of the levels of serum hormone, hormone receptors, and DNMTs induced by BPA in F1 females at PND 21 and PND 56. Forty milligrams per kilogram of CCFs significantly inhibited the hypermethylation of the H19/Igf2 imprinted gene induced by BPA (P < 0.01). It indicated that CCFs adjusted H19/Igf2 methylation by increasing the expression of DNMTs, thereby increasing the levels of reproductive hormones and receptors along with reducing the cytoapoptosis.

Maternal exposure to bisphenol A during pregnancy interferes testis development of F1 male mice.

This study was conducted to investigate the effects of maternal exposure to bisphenol A (BPA) on testis development of F1 male mice. The BPA exposure model of pregnant mice was prepared by intragastric administration of BPA at the doses of 0, 2.5, 5, 10, 20, and 40 mg/kg/day at gestation day (GD) 0.5-17.5. The testis index of the offspring mice was calculated at postnatal day (PND) 21 and PND 56. The results showed that maternal exposure to 20 mg/kg BPA during pregnancy significantly increased the testicular index of F1 males at PND 21, and 40 mg/kg BPA significantly decreased the testicular index of F1 males at PND 56 (P < 0.01). BPA significantly reduced serum testosterone (T) and estradiol (E2) levels, and improved testicular ERα and ERß levels in F1 males at both PND 21 and PND 56. BPA exposure also upregulated transcription of testicular Dnmt1 and inhibited the transcription of testicular Dnmt3A and Dnmt3B in F1 mice at PND 21. BPA reduced the transcriptional level of testicular DNA methyltransferase (Dnmt), increased the expression of testicular caspase-7, caspase-9, and bax, and decreased the expression of bcl-2 in F1 mice at PND 56. Consistent with that, BPA improved the apoptosis rate in the testis at PND 56 (P < 0.01 or P < 0.05). Our study indicates that BPA disrupts the secretion of testosterone, estradiol, and estrogen receptors by interfering with the transcription of testicular DNA methyltransferase (Dnmt) in offspring males, which damages testicular tissues and affects the potential reproductive function.