摘要:
BACKGROUND: Pulmonary fibrosis is a fatal lung disease with limited treatment options. Icaritin is the active ingredient derived from the traditional Chinese medical plant Epimedium and possesses many biomedical activities. This study aimed to investigate the effects and molecular mechanisms of icaritin on bleomycin-induced pulmonary fibrosis in mice. METHODS: To assess its preventative effects, bleomycin treated mice received 0, 0.04, 0.2, and 1 mg/kg of icaritin from day 1 onwards. To assess its therapeutic effects, bleomycin treated mice received 0 and 1 mg/kg of icaritin from day 15 onwards. Mice were sacrificed on day 21 and lung tissues were collected, stained with HE, Masson and immunohistochemistry. Q-PCR was used to measure Collagen I and Collagen III expression, western blotting was used to quantify α-SMA, Collagen I expression. Hydroxyproline content was measured using a biochemical method. NIH3T3 and HLF-1 cells were treated with TGF-β1with or without icaritin, and α-SMA, Collagen I were tested. PPARγ antagonist GW9662 and PPARγ-targeted siRNA were used to investigate the mechanism of icaritin in inhibiting myofibroblast differentiation. RESULTS: Both preventative and therapeutic administration of icaritin improved the histopathological changes, decreased Collagen and α-SMA, lowered hydroxyproline content in bleomycin-treated lung tissues. Icaritin decreased α-SMA and Collagen I expression in TGF-β1-stimulated NIH3T3 and HLF-1 cells. However, its effect in reducing α-SMA and Collagen I expression was suppressed when expression or activity of PPARγ was inhibited. CONCLUSIONS: Icaritin has therapeutic potential against pulmonary fibrosis via the inhibition of myofibroblast differentiation, which may be mediated by PPARγ.
期刊:
Medical and Veterinary Entomology,2021年35(1):42-50 ISSN:0269-283X
通讯作者:
Cheng, T. -Y.
作者机构:
[Tang, H.; Liu, L.; Cheng, T. -Y.; Feng, L. -L.] Hunan Agr Univ, Coll Vet Med, Hunan Collaborat Innovat Ctr Safety Prod Livestoc, Hunan Prov Key Lab Prot Engn Anim Vaccines, Changsha, Peoples R China.;[Cheng, T. -Y.] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Peoples R China.
通讯机构:
[Cheng, T. -Y.] H;Hunan Agr Univ, Coll Vet Med, Changsha 410128, Peoples R China.
关键词:
Anticoagulation;full-length cDNA;prokaryotic expression;recombinant protein
摘要:
Hemalin, initially described in Haemaphysalis longicornis, is a protein with anticoagulant activity. We retrieved a gene fragment functionally annotated as hemalin from H. flava salivary gland transcriptomic library, but its full-length complementary DNA (cDNA) and antithrombogenicity have not been investigated in the species. Here we cloned the full length of hemalin (Hf-hemalin) by 3'-end rapid-amplification of cDNA ends, and the open reading frame (ORF) of Hf-hemalin was expressed in Escherichia coli. The recombinant protein (rHf-Hemalin) was tested for antithrombogenicity. The full-length of Hf-hemalin was 607 bp with an ORF of423 bp. Protein encoded by Hf-hemalin was predicted to contain 2 Kunitz domains and a signal peptide. The expression of Hf-hemalin in salivary glands, midguts and ovaries was higher in the semi-engorged than the fully engorged. Prokaryotic expression yielded a product of 40 kDa containing a glutathione S-transferase (GST) tag. Incubation of rHf-Hemalin with rat plasma significantly extended prothrombin time and activated partial thromboplastin time compared with normal saline and GST controls. Our data demonstrated that Hemalin from H. flava shared a similar primary structure with that from H. longicornis, and was also anticoagulant. Further investigations are needed to test its feasibility to be an antigen candidate for the development of vaccines against ticks.
通讯机构:
[Lixin Wen] C;Colleges of Veterinary Medicine, Hunan Agricultural University, Changsha, 410128, China<&wdkj&>Hunan Co-innovation Center of Animal Production Safety, Changsha, 410128, China
关键词:
Antiviral activity;PK15 cells;Porcine circovirus 2;Tyrosine kinase inhibitor STI-571 and PP2;Virus like particles
摘要:
Porcine circovirus type 2 (PCV2) causes huge economic losses in the global swine industry and has a complex and poorly understood virus-host interaction mechanism. We reported that the C-terminal of the capsid protein of all PCV2 isolates shared a strictly conserved PXXP motif that may interact with SH3 domain-containing tyrosine kinases; however, its roles in PCV2 cell entry and replication remain unknown. In this study, we determined that mRNA levels of two SH3 domain-containing tyrosine kinases family (Abl and Src) had distinct profiles (wild-type and PXXP-mutated) during PCV2 infections of PK15 cells. Therefore, we hypothesized that activities of tyrosine kinases (Abl and Fyn) in PK15 cells may be hijacked by PCV2 via its PXXP motif of the Cap, to favor virus replication. Specific inhibitors PP2 of Lck/Fyn and STI-571 of Abl family kinases decreased viral production through suppression of DNA and Cap synthesis at the replication stage. However, based on indirect immunofluorescence assay (IFA), entry of PCV2 virus-like particles (VLPs) into PK15 cells was not altered. Elucidating mechanisms of PCV2-host interactions should provide new insights for development of new compounds to prevent or reduce PCV2 infections.
摘要:
Ganoderma lucidum polysaccharide (GLP) extracted from Ganoderma lucidum (Leyss. ex Fr.) Karst, a traditional Chinese medicine, is a biologically active substance reported to possess anti-oxidative, anti-apoptotic, and neurological protection. However, it is unknown whether GLP have any protective effect against high-fat constituents-induced epithelial cell injury. The aim of this study was to investigate the protection and molecular mechanism of GLP on injury induced by palmitic acid (PA) in the intestinal porcine epithelial cell line (IPEC-J2). First, we tested whether the treatment of GLP attenuate PA-induced IPEC-J2 cell death. GLP markedly blocked PA-caused cytotoxicity and apoptosis in IPEC-J2 cells. Moreover, GLP recovered the decreased mitochondrial function and inhibited activation of caspase-dependent apoptotic pathway. Interestingly, PA promoted cell apoptosis and autophagy through stimulation of phosphorylation of mitogen-activated protein kinases (MAPKs), AMP-activated protein kinase (AMPK), and inhibition of phosphorylation of Akt and mammalian target of rapamycin (mTOR), which was reversed by GLP. Taken together, this study revealed a protective effect of GLP against PA-evoked IPEC-J2 cell death through anti-apoptotic and anti-autophagic properties.
摘要:
Sanguinarine (SA) and chelerythrine (CHE) are the main active components of the phytogenic livestock feed additive, Sangrovit(R). However, little information is available on the pharmacokinetics of Sangrovit(R) in poultry. The goal of this work was to study the pharmacokinetics of SA, CHE, and their metabolites, dihydrosanguinarine (DHSA) and dihydrochelerythrine (DHCHE), in 10 healthy female broiler chickens following oral (p.o.) administration of Sangrovit(R) and intravenous (i.v.) administration of a mixture of SA and CHE. The plasma samples were processed using two different simple protein precipitation methods because the parent drugs and metabolites are stable under different pH conditions. The absorption and metabolism of SA following p.o. administration were fast, with half-life (t1/2 ) values of 1.05 +/- 0.18 hr and 0.83 +/- 0.10 hr for SA and DHSA, respectively. The maximum concentration (Cmax ) of DHSA (2.49 +/- 1.4 mug/L) was higher that of SA (1.89 +/- 0.8 mug/L). The area under the concentration vs. time curve (AUC) values for SA and DHSA were 9.92 +/- 5.4 and 6.08 +/- 3.49 ng/ml hr, respectively. Following i.v. administration, the clearance (CL) of SA was 6.79 +/- 0.63 (L.h(-1) .kg(-1) ) with a t1/2 of 0.34 +/- 0.13 hr. The AUC values for DHSA and DHCHE were 7.48 +/- 1.05 and 0.52 +/- 0.09 (ng/ml hr), respectively. These data suggested that Sangrovit(R) had low absorption and bioavailability in broiler chickens. The work reported here provides useful information on the pharmacokinetic behavior of Sangrovit(R) after p.o. and i.v. administration in broiler chickens, which is important for the evaluation of its use in poultry.
作者:
Wang, H. B.;Zhang, H. J.;Song, L. L.;Zhu, L.;Chen, M.;...
期刊:
JOURNAL OF HELMINTHOLOGY,2019年93(5):601-607 ISSN:0022-149X
通讯作者:
Zhao, G. H.;Liu, G. H.
作者机构:
[Chen, M.; Zhu, L.; Zhang, H. J.; Zhao, G. H.; Wang, H. B.; Ren, G. J.] Northwest A&F Univ, Coll Vet Med, Yangling 712100, Shaanxi, Peoples R China.;[Wang, H. B.] China Agr Vet Biol Sci & Technol Co Ltd, Lanzhou 730046, Gansu, Peoples R China.;[Liu, G. H.; Song, L. L.] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.
通讯机构:
[Zhao, G. H.] N;[Liu, G. H.] H;Northwest A&F Univ, Coll Vet Med, Yangling 712100, Shaanxi, Peoples R China.;Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.
摘要:
The golden snub-nosed monkey (Rhinopithecus roxellana) is an endangered species endemic to China. Relatively little is known about the taxonomic status of soil-transmitted helminths (STH) in these monkeys. Trichuris spp. (syn. Trichocephalus) are among the most important STHs, causing significant socio-economic losses and public health concerns. To date, five Trichuris species have been reported in golden monkeys, including a novel species, T. rhinopiptheroxella, based on morphology. In the present study, molecular and morphological analysis was conducted on adult Trichuris worms obtained from a dead golden snub-nosed monkey, to better understand their taxonomic status. Morphology indicated that the adult Trichuris worms were similar to T. rhinopiptheroxella. To further ascertain their phylogenetic position, the complete mitochondrial (mt) genome of these worms was sequenced and characterized. The mt genome of T. rhinopiptheroxella is 14,186 bp, encoding 37 genes. Phylogenetic analysis based on the concatenated amino acids of 12 protein-coding genes (with the exception of atp8) indicated that T. rhinopiptheroxella was genetically distinct and exhibited 27.5-27.8% genetic distance between T. rhinopiptheroxella and other Trichuris spp. Our results support T. rhinopiptheroxella as a valid Trichuris species and suggest that mt DNA could serve as a marker for future studies on the classification, evolution and molecular epidemiology of Trichuris spp. from golden snub-nosed monkeys.
通讯机构:
[Wu, J; Sun, ZL] H;Hunan Agr Univ, Coll Vet Med, Dept Clin Vet Med, Changsha 410128, Hunan, Peoples R China.;Hunan Coinnovat Ctr Utilizat Bot Funct Ingredient, Changsha 410128, Hunan, Peoples R China.
关键词:
H2O2;IPEC-J2 cells;apoptosis;koumine
摘要:
Medicinal herbal plants have been commonly used for intervention in different diseases and improvement of health worldwide. Koumine, an alkaloid monomer found abundantly in Gelsemium plants, can be effectively used as an antioxidant. The purpose of this study was to evaluate the potential protective effect of koumine against hydrogen peroxide (H(2)O(2))-induced oxidative stress and apoptosis in porcine intestinal epithelial cell line (IPEC-J2 cells). MTT assays showed that koumine significantly increased cell viability in H(2)O(2)-mediated IPEC-J2 cells. Preincubation with koumine ameliorated H(2)O(2)-medicated apoptosis by decreasing reactive oxygen species (ROS) production, and efficiently suppressed the lactate dehydrogenase (LDH) release and malondialdehyde (MDA) production. Moreover, a loss of superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) activities was restored to normal level in H(2)O(2)-induced IPEC-J2 cells upon koumine exposure. Furthermore, pretreatment with koumine suppressed H(2)O(2)-mediated loss of mitochondrial membrane potential, caspase-9 and caspase-3 activation, decrease of Bcl-2 expression and elevation of Bax expressions. Collectively, the results of this study indicated that koumine possesses the cytoprotective effects in IPEC-J2 cells during exposure to H(2)O(2) by suppressing production of ROS, inhibiting the caspase-3 activity and influencing the expression of Bax and Bcl-2. Koumine could potentially serve as a protective effect against H(2)O(2)-induced apoptosis.
作者机构:
[Zou, Feng-Cai] Yunnan Agr Univ, Key Lab Vet Publ Hlth Higher Educ Yunnan Prov, Coll Vet Med, Vet Parasitol, Kunming, Yunnan, Peoples R China.;[Li, Zhao; Chang, Jiang-Yan; Lv, Yan; Yang, Jian-Fa] Yunnan Agr Univ, Key Lab Vet Publ Hlth Higher Educ Yunnan Prov, Coll Vet Med, Kunming, Yunnan, Peoples R China.;[Li, Zhao; Zhu, Xing-Quan] Chinese Acad Agr Sci, State Key Lab Vet Etiol Biol, Lanzhou Vet Res Inst, 1 Xujiaping, Lanzhou 730046, Gansu, Peoples R China.;[Liu, Guo-Hua] Hunan Agr Univ, Coll Vet Med, Changsha, Hunan, Peoples R China.
通讯机构:
[Zhu, Xing-Quan] C;Chinese Acad Agr Sci, State Key Lab Vet Etiol Biol, Lanzhou Vet Res Inst, 1 Xujiaping, Lanzhou 730046, Gansu, Peoples R China.
摘要:
We performed a molecular survey for Cytauxzoon felis infection in 311 domestic cats in Yunnan Province, China, in 2016 and found a prevalence of 21.5%. C. felis infection in domestic and wild cats in other provinces should be investigated to determine parasite prevalence and genetic diversity among cats throughout China.
摘要:
T-2 and HT-2 toxins can cause cytotoxicity and oxidative stress in animals, while DL-Selenomethionine plays an important role in preventing oxidative stress and improving cell viability. However, the role of DL-Selenomethionine in T-2/HT-2 toxins-induced cell damage is still unknown. In this study, we investigated whether DL-Selenomethionine plays a protective role against T-2/HT-2-induced cytotoxicity and oxidative stress in primary hepatocytes. Our results demonstrated that T-2/HT-2 toxins-exposed broiler hepatocytes exhibited significantly decreased cell viability and intracellular glutathione (GSH) concentration while increased Lacate dehydrogenase (LDH) leakage, intracellular reactive oxygen species (ROS), glutathione peroxidase (GSH-PX), malondialdehyde (MDA) and catalase (CAT) levels, as well as elevated expression levels of genes related to oxidative stress, in a toxin dose-dependent manner (P < 0.05). However, the application of DL-Selenomethionine into T-2/HT-2 treated hepatocytes effectively alleviated the adverse effects of T-2/HT-2, as demonstrated by increased cell viability, decreased LDH leakage, declined intracellular ROS and MDA levels, increased expression of oxidative stress-related genes, as well as accordingly enhanced activities of GSH, GSH-PX, SOD and CAT as compared to the control groups (P < 0.05). Therefore, our in vitro data demonstrate that DL-Selenomethionine can function as an effectively protective agent against T-2/HT-2-induced cytotoxicity and oxidative stress.
摘要:
Ticks and tick-borne diseases are of major public health concern. Currently, development of vaccines against ticks is considered crucial for their control. A critical step in this process is the screening of viable antigens. Faeces are byproducts of digestion and blood meal utilization, and partly reflect the vitality and vector potential of ticks. However, an integrated analysis of proteins in tick faeces is lacking. The present study explored the protein components in the faeces of the tick Haemaphysalis flava, by liquid chromatography–tandem mass spectrometry (LC/MS-MS) to identify potential protein antigens for vaccine development against ticks. Faeces from adult H. flava engorged females were collected. Proteins were extracted from faeces, and the trypsin-digested peptides were analyzed by LC/MS-MS. High confidence proteins were identified based on unique peptides revealed by MS. Potential faecal protein genes, as well as their sources, were also characterized by searching previous transcriptome datasets from the salivary glands and midgut of H. flava. In total, 21 were recognized with confidence. Amongst these, 18 were of likely tick origin, while three proteins (serum albumin, haemoglobin α and β subunits) were likely from hosts. Seventeen unigenes corresponding to these proteins were retrieved by searching our previous H. flava salivary glands and midgut transcriptomic datasets. Some proteins were reported to prevent blood clotting, play a role in immunity and antibiosis, and formation of musculature. The functions of the remaining proteins are unknown. Identifying antigens for tick vaccine development is feasible by analyzing the faecal proteome as well as the transcriptomes of salivary glands and midguts. The vast number of proteins detected in tick faeces highlights the complexity of blood digestion in ticks, a field that needs more investigation.