Background: Granulosa cells(GCs) proliferation and estradiol synthesis significantly affect follicular development.The miR-214-3p expression in the ovarian tissues of high-yielding sows is higher than that in low-yiel...Background: Granulosa cells(GCs) proliferation and estradiol synthesis significantly affect follicular development.The miR-214-3p expression in the ovarian tissues of high-yielding sows is higher than that in low-yielding sows,indicating that miR-214-3p may be involved in sow fertility. However, the functions and mechanisms of miR-214-3p on GCs are unclear. This study focuses on miR-214-3p in terms of the effects on GCs proliferation and estradiol synthesis.Results: Our findings revealed that miR-214-3p promotes proliferation and inhibits estradiol synthesis in porcine GCs. MiR-214-3p can increase the percentage of S-phase cells, the number of EdU labeled positive cells, and cell viability. However, E2 concentration was reduced after miR-214-3p agomir treatment. We also found that miR-214-3p up-regulates the expression of cell cycle genes including cell cycle protein B(Cyclin B), cell cycle protein D(Cyclin D), cell cycle protein E(Cyclin E), and cyclin-dependent kinase 4(CDK4) at the transcription and translation levels, but down-regulates the mRNA and protein levels of cytochrome P450 family 11 subfamily A member 1(CYP11A1), cytochrome P450 family 19 subfamily A member 1(CYP19A1), and steroidogenic acute regulatory protein(StAR)(i.e., the key enzymes in estradiol synthesis). On-line prediction, bioinformatics analysis, a luciferase reporter assay, RT-qPCR, and Western blot results showed that the target genes of miR-214-3p in proliferation and estradiol synthesis are Mfn2 and NR5A1, respectively.Conclusions: Our findings suggest that miR-214-3 p plays an important role in the functional regulation of porcine GCs and therefore may be a target gene for regulating follicular development.展开更多
Background Clock circadian regulator(CLOCK)is a core factor of the mammalian biological clock system in regulat-ing female fertility and ovarian physiology.However,CLOCK’s specific function and molecular mechanism in...Background Clock circadian regulator(CLOCK)is a core factor of the mammalian biological clock system in regulat-ing female fertility and ovarian physiology.However,CLOCK’s specific function and molecular mechanism in porcine granulosa cells(GCs)remain unclear.In this study,we focused on CLOCK’s effects on GC proliferation.Results CLOCK significantly inhibited cell proliferation in porcine GCs.CLOCK decreased the expression of cell cycle-related genes,including CCNB1,CCNE1,and CDK4 at the mRNA and protein levels.CDKN1A levels were upregulated by CLOCK.ASB9 is a newly-identified target of CLOCK that inhibits GC proliferation;CLOCK binds to the E-box element in the ASB9 promoter.Conclusions These findings suggest that CLOCK inhibits the proliferation of porcine ovarian GCs by increasing ASB9 level.展开更多
Background:Oleic acid is an abundant free fatty acid present in livestock that are in a negative energy-balance state,and it may have detrimental effects on female reproduction and fertility.Oleic acid induces lipid a...Background:Oleic acid is an abundant free fatty acid present in livestock that are in a negative energy-balance state,and it may have detrimental effects on female reproduction and fertility.Oleic acid induces lipid accumulation in bovine granulosa cells,which leads to a foam cell-like morphology and reduced steroidogenesis.However,why oleic acid increases lipid accumulation but decreases steroidogenesis remains unclear.This study focused on oleic acid’s effects on lipid type and steroidogenesis.Results:Oleic acid increased the lipid accumulation in a concentration-dependent manner and mainly increased the triglyceride level and decreased the cholesterol ester level.Oleic acid also led to a decline in estradiol and progesterone production in porcine granulosa cells in vitro.In addition,oleic acid up-regulated the expression of CD36 and diacylglycerol acyltransferase 2,but down-regulated the expression of 3-hydroxy-3-methylglutarylcoenzyme A reductase,scavenger receptor class B member 1 and acetyl-Coenzyme A acetyltransferase 2,as well as steroidogenesis-related genes,including cytochrome P450 family 11 subfamily A member 1,cytochrome P450family 19 subfamily A member 1 and 3 as well as steroidogenic acute regulatory protein at the mRNA and protein levels.An oleic acid-rich diet also enhanced the triglyceride levels and reduced the cholesterol levels in ovarian tissues of female mice,which resulted in lower estradiol levels than in control-fed mice.Compared with the control,decreases in estrus days and the numbers of antral follicles and corpora lutea,as well as an increase in the numbers of the atretic follicles,were found in the oleic acid-fed female mice.Conclusions:Oleic acid changed the lipid type stored in lipid droplets of ovarian granulosa cells,and led to a decrease in steroidogenesis.These results improve our understanding of fertility decline in livestock that are in a negative energy-balance state.展开更多
The widespread use of feed additives in intensive and large-scale pig farming has resulted in high levels of heavy metals in pig manure.The long-term application of organic fertilizers containing high levels of heavy ...The widespread use of feed additives in intensive and large-scale pig farming has resulted in high levels of heavy metals in pig manure.The long-term application of organic fertilizers containing high levels of heavy metals leads to the accumulation of heavy metals in the soil,which not only causes heavy metal pollution in the soil,and also affect food safety and endanger human health.Composting is an economical and effective technical measures to achieve environmentally-sustainable treatment of pig manure and is a practical method to reduce the problem of heavy metals and to improve the resource value of pig manure.The composting process is accompanied by high temperatures and the production and emission of gases,and also lead to changes in the nitrogen content of the compost and provide opportunity for heavy metal passivation additives.This paper summarizes the forms and types of heavy metals present in pig manure and reviews the progress of research as well as the techniques and problems of in the composting process,and provides recommendations for research on heavy metal passivation and nitrogen retention in pig manure composting.展开更多
Skeletal muscle plays a paramount role in physical activity,metabolism,and energy balance,while its homeostasis is being challenged by multiple unfavorable factors such as injury,aging,or obesity.Exosomes,a subset of ...Skeletal muscle plays a paramount role in physical activity,metabolism,and energy balance,while its homeostasis is being challenged by multiple unfavorable factors such as injury,aging,or obesity.Exosomes,a subset of extracellular vesicles,are now recognized as essential mediators of intercellular communication,holding great clinical potential in the treatment of skeletal muscle diseases.Herein,we outline the recent research progress in exosomal isolation,characterization,and mechanism of action,and emphatically discuss current advances in exosomes derived from multiple organs and tissues,and engineered exosomes regarding the regulation of physiological and pathological development of skeletal muscle.These remarkable advances expand our understanding of myogenesis and muscle diseases.Meanwhile,the engineered exosome,as an endogenous nanocarrier combined with advanced design methodologies of biomolecules,will help to open up innovative therapeutic perspectives for the treatment of muscle diseases.展开更多
基金supported by grants from the National Natural Science Foundation (No.31802047)the National Science and Technology Major Project of China (No. 2016ZX08006003)Shaanxi Provincial Key Research and Development Project (CN)(No. 2018ZDXM-NY-035)。
文摘Background: Granulosa cells(GCs) proliferation and estradiol synthesis significantly affect follicular development.The miR-214-3p expression in the ovarian tissues of high-yielding sows is higher than that in low-yielding sows,indicating that miR-214-3p may be involved in sow fertility. However, the functions and mechanisms of miR-214-3p on GCs are unclear. This study focuses on miR-214-3p in terms of the effects on GCs proliferation and estradiol synthesis.Results: Our findings revealed that miR-214-3p promotes proliferation and inhibits estradiol synthesis in porcine GCs. MiR-214-3p can increase the percentage of S-phase cells, the number of EdU labeled positive cells, and cell viability. However, E2 concentration was reduced after miR-214-3p agomir treatment. We also found that miR-214-3p up-regulates the expression of cell cycle genes including cell cycle protein B(Cyclin B), cell cycle protein D(Cyclin D), cell cycle protein E(Cyclin E), and cyclin-dependent kinase 4(CDK4) at the transcription and translation levels, but down-regulates the mRNA and protein levels of cytochrome P450 family 11 subfamily A member 1(CYP11A1), cytochrome P450 family 19 subfamily A member 1(CYP19A1), and steroidogenic acute regulatory protein(StAR)(i.e., the key enzymes in estradiol synthesis). On-line prediction, bioinformatics analysis, a luciferase reporter assay, RT-qPCR, and Western blot results showed that the target genes of miR-214-3p in proliferation and estradiol synthesis are Mfn2 and NR5A1, respectively.Conclusions: Our findings suggest that miR-214-3 p plays an important role in the functional regulation of porcine GCs and therefore may be a target gene for regulating follicular development.
基金supported by National Natural Science Foundation of China (32272849)China Agriculture Research System of MOF and MARA
文摘Background Clock circadian regulator(CLOCK)is a core factor of the mammalian biological clock system in regulat-ing female fertility and ovarian physiology.However,CLOCK’s specific function and molecular mechanism in porcine granulosa cells(GCs)remain unclear.In this study,we focused on CLOCK’s effects on GC proliferation.Results CLOCK significantly inhibited cell proliferation in porcine GCs.CLOCK decreased the expression of cell cycle-related genes,including CCNB1,CCNE1,and CDK4 at the mRNA and protein levels.CDKN1A levels were upregulated by CLOCK.ASB9 is a newly-identified target of CLOCK that inhibits GC proliferation;CLOCK binds to the E-box element in the ASB9 promoter.Conclusions These findings suggest that CLOCK inhibits the proliferation of porcine ovarian GCs by increasing ASB9 level.
基金supported by grants from the National Natural Science Foundation(No.31802047)the National Science and Technology Major Project of China(No.2016ZX08006003)。
文摘Background:Oleic acid is an abundant free fatty acid present in livestock that are in a negative energy-balance state,and it may have detrimental effects on female reproduction and fertility.Oleic acid induces lipid accumulation in bovine granulosa cells,which leads to a foam cell-like morphology and reduced steroidogenesis.However,why oleic acid increases lipid accumulation but decreases steroidogenesis remains unclear.This study focused on oleic acid’s effects on lipid type and steroidogenesis.Results:Oleic acid increased the lipid accumulation in a concentration-dependent manner and mainly increased the triglyceride level and decreased the cholesterol ester level.Oleic acid also led to a decline in estradiol and progesterone production in porcine granulosa cells in vitro.In addition,oleic acid up-regulated the expression of CD36 and diacylglycerol acyltransferase 2,but down-regulated the expression of 3-hydroxy-3-methylglutarylcoenzyme A reductase,scavenger receptor class B member 1 and acetyl-Coenzyme A acetyltransferase 2,as well as steroidogenesis-related genes,including cytochrome P450 family 11 subfamily A member 1,cytochrome P450family 19 subfamily A member 1 and 3 as well as steroidogenic acute regulatory protein at the mRNA and protein levels.An oleic acid-rich diet also enhanced the triglyceride levels and reduced the cholesterol levels in ovarian tissues of female mice,which resulted in lower estradiol levels than in control-fed mice.Compared with the control,decreases in estrus days and the numbers of antral follicles and corpora lutea,as well as an increase in the numbers of the atretic follicles,were found in the oleic acid-fed female mice.Conclusions:Oleic acid changed the lipid type stored in lipid droplets of ovarian granulosa cells,and led to a decrease in steroidogenesis.These results improve our understanding of fertility decline in livestock that are in a negative energy-balance state.
基金supported by the National Key Research and Development Program of China(2018YFE0127000)Key R&D Program of Shaanxi Province(2022ZDLNY02-09)+1 种基金China Agriculture Research System(CARS-23-C-05)Postdoctoral Foundation of the Shaanxi Province(2018BSHEDZZ20)
文摘The widespread use of feed additives in intensive and large-scale pig farming has resulted in high levels of heavy metals in pig manure.The long-term application of organic fertilizers containing high levels of heavy metals leads to the accumulation of heavy metals in the soil,which not only causes heavy metal pollution in the soil,and also affect food safety and endanger human health.Composting is an economical and effective technical measures to achieve environmentally-sustainable treatment of pig manure and is a practical method to reduce the problem of heavy metals and to improve the resource value of pig manure.The composting process is accompanied by high temperatures and the production and emission of gases,and also lead to changes in the nitrogen content of the compost and provide opportunity for heavy metal passivation additives.This paper summarizes the forms and types of heavy metals present in pig manure and reviews the progress of research as well as the techniques and problems of in the composting process,and provides recommendations for research on heavy metal passivation and nitrogen retention in pig manure composting.
基金This work was supported by the National Key Research and Development Program of China(No.2021YFF1000602)。
文摘Skeletal muscle plays a paramount role in physical activity,metabolism,and energy balance,while its homeostasis is being challenged by multiple unfavorable factors such as injury,aging,or obesity.Exosomes,a subset of extracellular vesicles,are now recognized as essential mediators of intercellular communication,holding great clinical potential in the treatment of skeletal muscle diseases.Herein,we outline the recent research progress in exosomal isolation,characterization,and mechanism of action,and emphatically discuss current advances in exosomes derived from multiple organs and tissues,and engineered exosomes regarding the regulation of physiological and pathological development of skeletal muscle.These remarkable advances expand our understanding of myogenesis and muscle diseases.Meanwhile,the engineered exosome,as an endogenous nanocarrier combined with advanced design methodologies of biomolecules,will help to open up innovative therapeutic perspectives for the treatment of muscle diseases.
