Effect of Penthorum Chinense Pursh Compound on AFB1-Induced Immune Imbalance via JAK/STAT Signaling Pathway in Spleen of Broiler Chicken.

Aflatoxin B1(AFB1) is the main secondary metabolite produced by Aspergillus flavus, which is highly toxic, carcinogenic, mutagenic and teratogenic. It can induce immune imbalance in animals or humans. Penthorum chinense Pursh (PCP) is a traditional herbal plant that has been used as a hepatoprotective drug with a long history in China. Based on the theory of traditional Chinese Medicine, we prepared Penthorum chinense Pursh Compound (PCPC) by combining four herbal medicines: 5 g Penthorum chinense Pursh, 5 g Radix bupleuri, 1 g Artemisia capillaris Thunb and 1 g Radix glycyrrhizae. The role of the Penthorum chinense Pursh Compound (PCPC) in preventing AFB1-induced immune imbalance in broiler chickens was studied. A total of 180 broiler chickens were equally distributed in six groups: controls, AFB1, YCHD and high-, medium- and low-dose PCPC treatment groups. After 28 days, broilers were anesthetized, and serum spleen and thymus samples were collected for analysis. Results show that AFB1 significantly increased and decreased the relative organ weight of the spleen and thymus, respectively. Pathological section of hematoxylin/eosin (H&E) stained spleen sections showed that AFB1 resulted in splenic tissue damage. Both the serum levels of Immunoglobulin A (IgA) and Immunoglobulin G (IgG) were suppressed in the AFB1 group. IL-6 was elevated in the AFB1 group. The balance between pro-inflammatory cytokines (IFN-γ and IL-2) and anti-inflammatory cytokine (IL-4) was disturbed by AFB1. The apoptosis-related protein and JAK/STAT pathway-related gene expression indicated that AFB1-induced apoptosis via JAK/STAT pathway. PCPC has proven its immunoprotective effects by preventing AFB1-induced immune imbalance. PCPC can be applied as a novel immune-modulating medicine in broiler chickens. It can be applied as a novel immune modulator in veterinary clinical practice.

Penthorum chinense Pursh compound flavonoids supplementation alleviates Aflatoxin B1-induced liver injury via modulation of intestinal barrier and gut microbiota in broiler.

Aflatoxin B1 (AFB1) is a commonly occurring toxicant in animal and human diets, leading to hazardous effects on health. AFB1 is known to be a hepato-toxicant, and the intestinal barrier may play a crucial role in reversing AFB1-induced liver injury. This study aimed to optimize the extraction conditions of Penthorum chinense Pursh Compound Flavonoids (PCPCF) by the response surface method with a Box-Behnken design and investigate the effects of PCPCF on AFB1-induced liver injury in broilers. A total of 164 one-day-old broilers were divided into seven groups, including Control, PCPCF (400 mg PCPCF/kg feed), AFB1 (3 mg AFB1/kg feed), and YCHT (Yin-Chen-Hao-Tang extract, 3 mg AFB1 +10 mL YCHT/kg feed) and low, medium, and high dose groups (PCPCF at 3 mg AFB1 +200, 400, 600 mg respectively). Samples of serum, liver, duodenum, and cecum contents were collected at 14th and 28th days for further analysis. The results showed that the maximum extraction rate of PCPCF was 8.15 %. PCPCF was rich in rutin, quercetin, liquiritin and kaempferol, and significantly inhibited the growth of Aspergillus flavus. The addition of PCPCF improved the growth performance of AFB1-injury broilers, modulated liver function, and increased serum immunoglobulin levels. PCPCF also alleviated liver pathological and oxidative stress damages caused by AFB1 and decreased AFB1-DNA and AFB1-lysine content in the liver. Furthermore, PCPCF supplementation ameliorated intestinal pathological damage, improved intestinal permeability of duodenum in the AFB1-induced broilers, and repaired the intestinal mucosal and mechanical barrier associated with the Notch signaling pathway. Meanwhile, PCPCF improved the intestinal flora structure of AFB1-damaged broilers and increased the abundance of beneficial bacteria. In conclusion, PCPCF ameliorated the adverse effects of AFB1 on growth performance and alleviated liver damage by repairing the intestinal barrier and improving intestinal health of broiler chicken.

An Updated Review on Efficiency of Penthorum chinense Pursh in Traditional Uses, Toxicology, and Clinical Trials.

Traditional Chinese medicines (TCM) play an important role in the control and treatment of several animal diseases. Penthorum chinense Pursh (PCP) is a famous plant for its use in traditional medication practice and therapeutic effects in numerous pathological conditions. In China, PCP is utilized for both food and medication due to numerous bioactivities. PCP is widely administered in prevention and treatment of traumatic injury, edema, and liver diseases with functions of reducing swelling, support diuresis, blood stasis, and mitigation symptoms of excessive alcohol intake. Recently, PCP highlighted for research trials in various fields including pharmacology, pharmacognosy, cosmeceuticals, nutraceuticals, and pharmaceuticals due to medicinal significance with less toxicity and an effective ethnomedicine in veterinary practice. PCP contains diverse important ingredients such as flavonoids, organic acids, coumarins, lignans, polyphenols, and sterols that are important bioactive constituents of PCP exerting the therapeutic benefits and organ-protecting effects. In veterinary, PCP extract, compound, and phytochemicals/biomolecules significantly reversed the liver and kidney injuries, via antioxidation, oxidative stress, apoptosis, mitochondrial signaling pathways, and related genes. PCP water extract and compounds also proved in animal and humans' clinical trial for their hepatoprotective, antiaging, nephroprotective, anti-inflammatory, antidiabetic, antibacterial, antiapoptotic, immune regulation, and antioxidative stress pathways. This updated review spotlighted the current information on efficiency and application of PCP by compiling and reviewing recent publications on animal research. In addition, this review discussed the toxicology, traditional use, comparative, and clinical application of PCP in veterinary practices to authenticate and find out new perspectives on the research and development of this herbal medicine.

Effect of In Ovo Trace Element Supplementation on Immune-Related Cells of the Small Intestine of Post-hatched Broiler Chicken.

Pathological conditions and harmful drugs cause many gastrointestinal diseases in broiler chicken. The current study was conducted to investigate the effect of trace elements zinc (Zn) and selenium (Se) supplementation on histomorphology, immunological role, and functional activity of goblet cells (GCs) of the small intestine. The Alcian blue-periodic acid-Schiff (AB-PAS) was performed to assess the histomorphological changes in GCs, which revealed the regular dispersion with high electron density of GCs throughout the mucosal surface in the supplemented group. However, irregular dispersion with low electron density of GCs was present in the control group. The immunological functional role of GCs within the small intestine was examined by mucicarmine staining, immunohistochemistry, and immunofluorescence. The results showed a high mucin glycol protein secretion in the supplemented group, whereas limited mucin glycol protein secretion in the control group. Furthermore, the biological significance showed a high and low immunoreactivity of Muc2 and Muc13 in the supplemented and control groups, respectively. Immunofluorescence was used to confirm the immunosignaling of Muc2. Results revealed high immunosignaling of Muc2 at the apical part of the small intestine in the supplementation group, while low immunosignaling of Muc2 in the control group. Results suggest that trace element supplementation had significant effect on morphology and immunological role of GCs, which might be essential for immune function and health status of broiler chicken.

Effect of early feeding of L-arginine and L-threonine on hatchability and post-hatch performance of broiler chicken.

This study aimed to evaluate the effect of in ovo feeding (IOF) of L-arginine (Arg) and L-threonine (Thr) in the broiler. For this purpose, 500 embryonated eggs were randomly allocated into five treatment groups of four replicates 25 eggs/replicate. The five treatments were arranged as (1) non-injected control, (2) 0.75% NaCl injected group, (3) 25 mg/egg Arg 4) 25 mg/egg Thr, and (5) Arg + Thr25 mg/egg. On the 17th day of incubation, 0.5 ml of treatment solution was injected into the amniotic fluid of all treatment groups. The result showed that the supplementation group of Arg + Thr significantly (P < 0.05) improved the hatchability, post-hatch growth performance, organ weight, and organ development in compression to sham control and other treatment groups. The antibiotic titer of NDV was improved in Arg + Thr group. Moreover, hematological indices were improved significantly in Arg + Thr group. The plasma concentration of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were noted to decrease in Arg + Thr group. Histopathological investigation revealed that IOF of Arg + Thr increased the villi length and crypt depth of the intestine. Conclusively, the IOF Arg and Thr could be an effective way to optimize the health and productive performance of broilers.

Hepatoprotective Effect of the Penthorum Chinense Pursh Extract against the CCl4-Induced Acute Liver Injury via NF-κB and p38-MAPK PATHWAYS in Dogs.

Acute liver injury (ALI), manifested by acute hepatocellular damages and necrosis, is a life-threatening clinical syndrome and Penthorum Chinense Pursh (PCP) is a well-known folk medicine practiced for liver-related diseases. This study aimed to investigate the ameliorative effects of PCP extract (PCPE) on carbon tetrachloride (CCl4) induced ALI in dogs via mitogen-activated protein kinase (MAPK) and Nuclear factor κB (NF-κB) signaling pathway. Healthy dogs were induced by CCl4 and treated with different dosage regimes of PCPE for 7 days. CCl4 produced acute liver injury and induced both oxidative stress and an inflammatory response in dogs. The PCPE significantly ameliorated and improved vacuolar inflammatory lesions in liver tissues during ALI, enhanced activity of superoxide dismutase, and restored glutathione peroxidase, further significantly reducing the indices of malondialdehyde and nitric oxide in serum. Inflammatory factors (IL-1ß, IL-6, and TNF-α) were declined and anti-inflammatory factors (IL-10) were increased by the application of PCPE. PCPE treatment, down-regulated the MEKK4, MKK3, p38MAPK, MSK1, and NF-κB, and upregulated the IkB mRNA levels (p < 0.01) in ALI affected dogs. In conclusion, PCPE repaired acute liver injury by improving antioxidant enzymes and by reducing oxidation products. Furthermore, the PCPE inhibited the MAPK/NF-κB signaling pathway, which resulted in anti-inflammatory and antioxidant effects on ALI-induced dogs. In the future, PCPE could be a useful ethnomedicine in veterinary clinical practices for the treatment of liver injuries or failures.

Penthorum Chinense Pursh Extract Alleviates Aflatoxin B1-Induced Liver Injury and Oxidative Stress Through Mitochondrial Pathways in Broilers.

Aflatoxin is an important toxicant of the fungal origin and poses a threat to the poultry industry. This study was designed to reveal the underlying mechanism and protective methods against aflatoxin B1 (AFB1)-induced liver injury, oxidative stress, and apoptosis using a Traditional Chinese medicine, Penthorum chinense Pursh extract (PCPE), in broilers. A total of 164 (day-old) broilers were equally allocated to the control, AFB1 (3 mg/kg feed), positive drug (Yin-Chen-Hao Tang extract, 10 ml/kg feed), PCPE (2 g PCPE/kg), and PCPE low, medium, and high dose groups (1 g, 2 g, 3 g PCPE/kg feed, respectively). AFB1 significantly decreased the growth performance and serum immunoglobulin level, altered normal serum biochemical parameters and antioxidant activities, and induced histopathological lesions in the liver as compared to control group. Additionally, AFB1 significantly up-regulated the mRNA expression levels of apoptosis-related genes such as Bax, Bak, caspase-9, caspase-3, and p53, whereas it down-regulated the expression levels of BCL2 in the liver of broilers. The supplementation of different doses of PCPE to AFB1-affected birds significantly eased AFB1 negative effects by improving growth performance, immunoglobulin level, and oxidative capacity, and reversed oxidative stress and pathological lesions in liver. Furthermore, supplementation of PCPE to the AFB1 group reversed apoptosis by significantly down-regulating the mRNA expression levels of Bax, Bak, caspase-9, caspase-3, and p53 and up-regulating the expression levels of BCL2 in the liver of broilers. Based on these results, we conclude that supplementation of PCPE is protective and safe against oxidative stress, is anti-apoptotic, and reverses the liver damage caused by AFB1 in broilers.

Health benefits of carotenoids and potential application in poultry industry: A review.

Carotenoids are one of the widespread and ubiquitous lipid-soluble pigments that produce a wide range of colours which are universally found in various plants, microalgae, bacteria and fungi. Recently, interest in using carotenoids as feed ingredients has increased markedly owing to their bioactive and health-promoting properties. In terms of applications, carotenoid-rich products are widely available in the form of food and feed additive, supplements and natural colourants. Carotenoids play a versatile biological role that contributes to therapeutic effects, including anticancer, immunomodulators, anti-inflammatory, antibacterial, antidiabetic and neuroprotective. Dietary supplementation of carotenoids not only improves the production performance and health of poultry birds, but also enhances the quality of egg and meat. Several studies have suggested that the supplementation of plant derived carotenoids revealed numerous health-promoting activities in poultry birds. Carotenoids reduce the oxidative stress in pre-hatched and post-hatched birds through different mechanisms, including quench free radicals, activating antioxidant enzymes and inhibiting the signalling pathways. Use of carotenoids in poultry feed as a part of nutrient that confers bird health and improve product quality. Carotenoids play a critical role for the pigmentation of egg yolk, skin, legs, beak, comb, feather and fat. Birds consumed carotenoid deficient diet resulting hues of their egg yolk or pale coloured skin. Therefore, uniform pigmentation generally indicates the health status and quality of the poultry products. This review aims to gather recent information regarding bioactive properties of carotenoids and highlight pharmaceutical and health beneficial effects of carotenoids for the poultry industry. Additionally, it explores the importance of carotenoids as alternative feed ingredients for poultry to boost the production performance and replace synthetic medicine and nutrients.

Clinical efficiency and safety of Hsp90 inhibitor Novobiocin in avian tibial dyschondroplasia.

Tibial dyschondroplasia (TD) is a bone defect of broilers and other poultry birds that disturbs growth plate and it causes lameness. Previously we evaluated differential expression of multiple genes involved in growth plate angiogenesis and reported the safety and efficacious of medicinal plant root extracted for controlling TD. In this study, clinical and protective effect of an antibiotic Novobiocin (Hsp90 inhibitor) and expression of Hsp90 and proteoglycan aggrecan was examined. The chicks were divided into three groups; Control, thiram-induced TD, and Novobiocin injected TD. After the induction of TD, the Novobiocin was administered through intraperitoneal route to TD-affected birds until the end of the experiment. The expressions and localization of Hsp90 were evaluated by qRT-PCR, immunohistochemistry (IHC) and western blot, respectively. Morphological, histological examinations, and serum biomarker levels were evaluated to assess specificity and protective effects of Novobiocin. The results showed that TD causing retarded growth, enlarged growth plate, distended chondrocytes, irregular columns of cells, decreased antioxidant capacity, reduced protein levels of proteoglycan aggrecan, and upregulated in Hsp90 expression (p < 0.05) in dyschondroplastic birds as compared with control. Novobiocin treatment restored growth plate morphology, reducing width, stimulated chondrocyte differentiation, sprouting blood vessels, corrected oxidative imbalance, decreased Hsp90 expressions and increased aggrecan level. Novobiocin treatment controlled lameness and improved growth in broiler chicken induced by thiram. In conclusion, the accumulation of the cartilage and up-regulated Hsp90 are associated with TD pathogenesis and irregular chondrocyte morphology in TD is along with reduced aggrecan levels in the growth plate. Our results indicate that Novobiocin treatment has potential to reduce TD by controlling the expression of Hsp90 in addition to improve growth and hepatic toxicity in broiler chicken.

Icariin Ameliorate Thiram-Induced Tibial Dyschondroplasia via Regulation of WNT4 and VEGF Expression in Broiler Chickens.

Tibial dyschondroplasia (TD) is main bone problem in fast growing poultry birds that effect proximal growth plate (GP) of tibia bone. TD is broadly defined as non-vascularized and non-mineralized, and enlarged GP with tibia bone deformation and lameness. Icariin (Epimedium sagittatum) is a traditional Chinese medicine, which is commonly practiced in the treatment of various bone diseases. Recently, many researcher reports about the beneficial effects of icariin in relation to various types of bone conditions but no report is available about promoting effect of icariin against TD. Therefore, current study was conducted to explore the ameliorating effect of icariin in thiram-induced TD chickens. A total of 180 broiler chicks were equally distributed in three groups; control, TD induced by thiram (50 mg/kg), and icariin group (treated with icariin @10 mg/kg). All groups were administered with normal standard diet ad libitum regularly until the end of experiment. The wingless-type member 4 (WNT4) and vascular endothelial growth factor (VEGF) genes and proteins expression were analyzed by quantitative real-time polymerase chain reaction and western blot analysis respectively. Tibial bone parameters, physiological changes in serum, antioxidant enzymes, and chicken growth performance were determined to assess advantage and protective effect of the medicine in broiler chicken. The expression of WNT4 was decreased while VEGF increased significantly (P < 0.05) in TD affected chicks. TD enhanced the GP, lameness, and irregular chondrocytes, while reduced the liver function, antioxidant enzymes in liver, and performance of chickens. Icariin treatment up-regulated WNT4 and down-regulated VEGF gene and protein expressions significantly (P < 0.05), restored the GP width, increased growth performance, corrected liver functions and antioxidant enzymes levels in liver, and mitigated the lameness in broiler chickens. In conclusion, icariin administration recovered GP size, normalized performance and prevented lameness significantly. Therefore, icariin treatments are encouraged to reduce the incidence of TD in broiler chickens.

In Vitro Effect of Apigenin and Danshen in Tibial Dyschondroplasia Through Inhibition of Heat-Shock Protein 90 and Vascular Endothelial Growth Factor Expressions in Avian Growth Plate Cells.

Tibial dyschondroplasia (TD) is one of the common skeletal abnormalities in fast-growing birds, and it is characterized by nonvascularized, unmineralized, and nonviable cartilage in the tibial growth plate that fails to form bone. The aim of this study was to check the in vitro effect of apigenin and danshen on heat-shock protein 90 (Hsp90) and vascular endothelial growth factor (VEGF) expressions in avian growth plate cells treated with sublethal concentration of thiram. Initially, chondrocytes from chicken growth plates were isolated on culturx ed medium with and without various concentration of thiram to determine the sublethal dose. Then, to check the effect of apigenin and danshen, the chondrocytes were treated first with a sublethal (2.5 µM) concentration of thiram and then with different doses (10, 20, 40, and 80 µM) of apigenin and danshen. The mRNA expression levels of Hsp90 and VEGF genes were evaluated by quantitative reverse transcription polymerase chain reaction (RT-qPCR). The results showed that the expression levels of Hsp90 and VEGF mRNA transcripts were increased significantly (P < 0.05) in thiram-treated chondrocytes culture medium up to 1.5-fold, whereas apigenin and danshen therapy to chondrocytes in culture medium significantly (P < 0.05) reduced the Hsp90 and VEGF expression levels. In conclusion, up-regulation of both (Hsp90 and VEGF) genes and damage to chondrocytes in culture medium caused by thiram can be restored by using apigenin and danshen. Therefore, apigenin and danshen therapies are suggested and encouraged as a promising approach to control TD in broiler chickens.

Hsp90 inhibitor celastrol reinstates growth plate angiogenesis in thiram-induced tibial dyschondroplasia.

Tibial dyschondroplasia (TD) is an important long bone defect of broiler chickens that disturbs the proximal growth plate and is characterized by non-vascularized cartilage, a distended growth plate and lameness. Celastrol, a medicinal root extract from the plant Tripterygium wilfordii, is reported widely as a well-known heat-shock protein 90 (Hsp90) inhibitor. Recently, Hsp90 inhibition in chondrocyte differentiation and growth-plate vascularization were effective in restoring the morphology of the growth plate. The present study was aimed at investigating Hsp90 inhibition in TD using celastrol. The broiler chicks were divided into three groups; Control; TD induced (40 mg/kg thiram) and celastrol treatment. Hsp90, vascular endothelial growth factor and Flk-1 expressions were evaluated by quantitative real-time polymerase chain reaction and the protein levels of Hsp90 were measured by Western blot analysis. Antioxidant enzymes were determined to assess the liver damage caused by thiram and the protective effects of the medicine were evaluated by levels of serum biomarkers. The expression levels of Hsp90 and vascular endothelial growth factor mRNA transcripts were increased while Flk-1 receptor was decreased in TD-affected chicks. Celastrol therapy inhibited Hsp90 mRNA and protein levels and up-regulated the expressions of receptor Flk-1 in TD-affected tibial growth plates significantly (P < 0.05) in addition to rectifying the damaging effects of thiram on the liver by decreasing the levels of aspartate aminotransferase, alanine aminotransferase and malondialdehyde and correcting the oxidative imbalance. In conclusion, administering celastrol to dyschondroplastic chicks prevented un-vascularized growth plate, lameness and reinstated angiogenesis. Celastrol may be efficacious for the treatment of TD through the inhibition of Hsp90 expression and limiting the liver damage caused by thiram in broiler chickens.