CN108841781A - Crocin is improving the application in porcine oocytes oxidation resistance and cleavage rates - Google Patents
Crocin is improving the application in porcine oocytes oxidation resistance and cleavage rates Download PDFInfo
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Abstract
The invention discloses crocins to improve the application in porcine oocytes oxidation resistance and cleavage rates.300~500ug/mL is added in oocyte in vitro maturation culture solution, maturing rate and cleavage rates can be improved in the especially crocin of 400ug/mL, it reduces intracellular ROS concentration and improves GSH concentration, and improve the mrna expression amount of antioxidant genes, this is meaningful to pig embryo in vitro culture, and Oocyte quality and cleavage rates needs further increase.
Description
Technical field
The present invention relates to the embryo in vitro reproduction technique of pig, specially crocin is improving the anti-oxidant energy of porcine oocytes
Application in power and cleavage rates.
Background technique
Improve pig embryo production in vitro efficiency be it is very important for current scientific domain, since pig is for xenogenesis
Transplanting and embryo cleavage model have very high medical value (Telugu et al., 2011), therefore, improve external
Embryo Production efficiency is widely paid close attention to.However, due to oocyte in vitro maturation system it is incomplete etc. it is many it is extraneous because
Element causes the efficiency for obtaining embryo to be in vitro lower than in vivo, to influence (the Seo-jin Park et of the Oocyte quality after maturation
al.,2017).So many laboratories in the whole world attempt to improve ovum by the way of adding microelement to in-vitro culture medium
The quality (Kim E et al., 2015) of mother cell.
The affected big factors of quality are oxidative stress to egg mother cell in maturation culture in vitro
(oxidative stress).Egg mother cell is set to generate active oxygen (ROS) from external high oxygen concentration, in an in vitro environment
It is easier the interference by active oxygen compared to vivo environment, as the increase of activity keto concentration in cell leads to lipid, protein
It is impaired with DNA, eventually lead to cell death (A.Rodriguez-Rodriguez et al., 2014).This may be also resulted in
Embryo's is undermined prevention early embryonic development.Therefore, addition such as beta -mercaptoethanol, half Guang in oocyte IVM base
It is impaired (Nakamura BN et al., 2011) from unfavorable factor bring that the substances such as propylhomoserin can protect cell.
Crocin (crocin) is the kermesinus column cap of the drying of plant crocus cultivation, is water-soluble class Hu trailing plants
Bu Su has the advantages that oral (Naess et al., 2006).Researchers have found it to its special pharmacological Quality Research
Anti-oxidant and antitumor action (Gholamnezhad Z et al., 2013).Crocin is a Carotenoids, and class is recklessly
Radish element can resist ROS and macromolecular bring oxidative stress and protect cell (Alaluf S et al., 2002).Crocin
In mono-/bis glycosyl enester these carotenoid can remove free radical, especially superoxide anion, so they can
Cell (Bors W et al., 1982) can be protected from oxidative stress.Glutathione (GSH) is reported as providing cell from oxygen
Change stress toxic condition in play critically important protective effect, especially in high oxygen concentration.Then, in egg mother cell
GSH concentration improve cause it is in vitro fertilization after improve porcine oocytes and masculonucleus formation (Nguyen VL et al.,
2009).Nowadays, crocin is widely used in the increment for studying inhibition cancer cell, diseases associated with inflammation and itself exempts from the field of medicine
Epidemic disease disease etc. (Hoshyar R et al., 2016).
Summary of the invention
It is an object of the invention to add the application in crocin raising porcine oocytes oxidation resistance and cleavage rates.
Although researcher always strives to improve the maturation in vitro rate of egg mother cell, again below cylinder mature rate,
It is low (Gil MA et al., 2010) which results in rates in vitro fertilization and blastocyst rate.The mature quality of egg mother cell is for embryo's
Early development is very crucial, so oocyte in vitro maturation system is in vitro fertilization most important.
ROS is generated in oxidation-reduction process in the cell, and destroys intracellular organelle in high concentration ROS
And structure, so egg mother cell internal oxidition stress be assess Oocyte quality important parameter (Jeon Y et al.,
2014).GSH concentration is also to assess the important parameter of oocyte maturation quality in egg mother cell, and GSH participates in intracellular a variety of mistakes
Journey, including protect from oxidative stress cell and adjust cellular redox metabolism (Luberda Z et al., 2005).
Crocin is a Carotenoids, and strong antioxidant, discovery crocin can be supported in different intracorporeal organ research
System is from free radical and oxidative stress bring adverse effect (Bandegi AR et al., 2014).In this trial, we
Detect the mRNA expression of the ROS and GSH concentration and maturation in vitro rate, cleavage rates and antioxidant genes in mature oocyte
Amount.Our test result shows that adding crocin can be improved GSH concentration in egg mother cell and remove ROS, anti-oxidant base
The mrna expression amount of cause also significantly improves.
The recent research about antioxidant is reported, adds zeaxanthin in in-vitro maturity of porcine oocytes culture solution
(zeaxanthin) maturation and intracellular concentration of GSH of oocyte nuclei are significantly improved, but upper and subsequent in the reduction of ROS concentration
Cleavage rates difference it is not significant (Seo-jin Park et al., 2017).Melatonin (melatonin) is also strong
Free radical scavenger and antioxidant add melatonin in in-vitro maturity of porcine oocytes culture solution and effectively raise
Embryonic development (Zhen CHEN et al., 2017) after maturing rate and lonely female activation.Canthaxanthin (canthaxanthin) is one
Kind is naturally present in the various intracorporal antioxidants of biology, adds in in-vitro maturity of porcine oocytes culture solution
The maturation and the development of embryo after lonely female activation of egg mother cell can be improved in canthaxanthin, improves intracellular concentration of GSH simultaneously
And ROS concentration is reduced, have adjusted expression (the A Taweechaipaisankul et of apoptosis-related genes and development related gene
al.,2016).Maturing rate, rate of fertilization can be improved as a kind of effective antioxidant in chlorogenic acid (chlorogenic acid)
And embryo development rate, the H prevented2O2Cause DNA fragmentation (Nguyen TVet al.,2017).ROS is probably derived from embryo
In tire metabolism or embryo's local environment, this may be unfavorable to embryonic development.Many antioxidants can slow down oxygen in reproductive process
Change stress, and improve the developmental rate (Kang et al., 2013) of embryo in an in vitro environment.In oocyte in vitro maturation
Flavone compound antioxidant is added in culture solution, the intracellular GSH concentration of 2cell and 4cell after lonely female activation mention
High and ROS concentration reduces (Kang et al., 2016), this may improve it is in vitro fertilization after cleavage rates.
The beneficial effects obtained by the present invention are as follows being：Crocin is added in oocyte in vitro maturation culture solution can be improved
Maturing rate and cleavage rates reduce intracellular ROS concentration and improve GSH concentration, and improve the mrna expression amount of antioxidant genes,
This is meaningful to pig embryo in vitro culture, and Oocyte quality and cleavage rates needs further increase.
Detailed description of the invention
Attached drawing is used to provide further understanding of the present invention, and constitutes part of specification, with reality of the invention
It applies example to be used to explain the present invention together, not be construed as limiting the invention.
Fig. 1 is that ROS fluorescence intensity histogram after various concentration crocin In-vitro maturation is added in the embodiment of the present invention；
Fig. 2 is that GSH fluorescence intensity histogram after various concentration crocin In-vitro maturation is added in the embodiment of the present invention；
Fig. 3 is mature oocyte gene mRNA expression amount in the embodiment of the present invention
Specific embodiment
Hereinafter, preferred embodiments of the present invention will be described, it should be understood that preferred embodiment described herein is only used
In the description and interpretation present invention, it is not intended to limit the present invention.
Embodiment
Pig ovary is collected
Pig ovary adds 37 DEG C of physiological saline (0.9%), 2h in local slaughterhouse, transportational process in vacuum flask
Within reach laboratory.Liquor folliculi is extracted with 20ml syringe, choosing diameter is 3-6mm ovarian follicle, and liquor folliculi is loaded on 15ml centrifuge tube
In be placed in 38.5 DEG C of water-baths to picking COCs.
Oocyte in vitro maturation test：
In each processing group, the COCs with three layers of granular cell is selected.Mature liquid be 1ml TCM-199 (M2520,
The Shanghai SIGMA branch company) it include 1ul/ml hCG, 1ul/ml PMSG, 1ul/ml Egf in culture solution.25-30 cell of picking
It is placed in each droplet (70ul) covering mineral oil, is divided into 0,300ug/ml, 400ug/ml and 500ug/ml processing group, 38.5
DEG C, 5%CO2, maximum saturation humidity incubator in cultivate 44h.
(IVF) in vitro fertilization and in vitro culture (IVC)
After oocyte maturation, the ovarian cumulus around egg mother cell is gently blown and beaten in hyaluronidase droplet with liquid-transfering gun
Cell is until being completely exposed oolemma.Then it is cleaned 3 times in capacitation liquid, be put into fertilization droplet (90ul) and cover mineral
Oily (each droplet is put into 20 egg mother cells).The fresh semen that laboratory is transported within the 2h of endemic species pig farm is dilute with capacitation liquid
It releases, ultimate density is 1 × 105cells/ml.With capacitation liquid in 38.5 DEG C, 5%CO2, maximum saturation humidity incubator in capacitation
Then 15min injects 10ul sperm, in 38.5 DEG C, 5%CO in each fertilization droplet2, maximum saturation humidity incubator in
(IVF) 4h in vitro fertilization.Egg mother cell is gently blown and beaten 3-5 times in embryo medium (NCSU-23) after 4h, female ovum is attached to
The extra sperm of cell oolemma washes.It is transferred to embryo medium droplet (500ul) and covers mineral oil (each droplet is put
Enter 20 egg mother cells), in 38.5 DEG C, 5%CO2, maximum saturation humidity incubator in cultivate.It is set as the 0th day on the day of IVF,
Cleavage rates are observed after 72h.It is spaced 48h replacement embryo medium (T.Somfai et al., 2010).
The measurement of oocyte maturation degree
After egg mother cell 44h maturation in vitro, in hyaluronidase droplet gently piping and druming wash surrounding ovarian cumulus it is thin
Born of the same parents clean 3-5 times in 0.1%PBS-PVA droplet later until oolemma completely reveals.Then at maximum times of microscope
The egg mother cell of picking discharge first polar body, discharge first polar body are mature oocyte in number.Calculate mature oocyte number
Amount accounts for the ratio of entire egg mother cell quantity.
The measurement of ROS and GSH fluorescence intensity
After egg mother cell 44h maturation in vitro, removes cumulus cell (details is same as above), detect intracellular ROS and GSH respectively
Concentration.By the method for You et al., respectively with dichlorohydrofluorescein diacetate (DCHFDA) and
CellTracker Blue CMF2HC (4-chloromethyl-6,8-difluoro-7-hydroxycoumarin) is detected
Fluorescence intensity (You et al., 2010).Each 20 egg mother cells are respectively in 10uM DCHFDA and 10uM Cell Tracker
38.5 DEG C, 5%CO2, 30min is cultivated in maximum saturation humidity environment incubator.It is clear in 0.1PBS-PVA droplet after culture
It washes 3-5 times, is installed in glass slide and covers coverslip.It is placed in fluorescence microscope (Lecia DM IRB, Niokn, Japan) and looks into
See (460nm for ROS and 370nm for GSH).Fluorescent image is analyzed with Image J software.
The measurement of antioxidant genes mRNA expression
After egg mother cell 44h maturation in vitro, useSuper RNA extracts kit (Promega, China
Shanghai), egg mother cell is cracked, the pollution of DNA enzymatic I Incubating Solution removal genomic DNA is then added, by RNA repeatedly
Washing lotion cleaning, the final RNA for obtaining high-purity.The RNA of extraction reverse transcription reagent box GoScriptTM Reverse
Transcription Mix, Oligo (dT) (Madison, WI 53711-5399, USA) add Nuclease-Free
Water 4ul,GoScriptTM Reaction Buffer,Oligo(dT)4ul,GoScriptTMEnzyme Mix 2ul into
Row reverse transcription, reverse transcription condition are 25 DEG C of 5min, and 42 DEG C of 60min, 70 DEG C of 15min finally cool to 4 DEG C, obtain cDNA.
Taq 6.5ul, H are added in 1ul cDNA2O 4ul, positive anti-primer respectively add 2.2ul, then add mineral oil covering
In order to avoid evaporation.Be put into PCR instrument and expand, amplification condition be 95 DEG C of 3min, 40 circulation 95 DEG C of 30s, 59 DEG C of 30s, 72 DEG C
30s, then 72 DEG C of 5min, finally cool to 4 DEG C.The sample obtained runs 25min electrophoresis in Ago-Gel.The film obtained is put
Enter imaging system to be analyzed in software LANE.Table 1 lists Primer, sequence and size.glyceraldehyde-3-
Phosphate dehydrogenase (GAPDH) gene is referred to as reference gene.
1 specific primer of table is expanded for genetic fragment using round pcr.
The measurement of oocyte fertilization rate
After egg mother cell 44h maturation in vitro, remove cumulus cell (details is same as above), be put into IVF droplet at capacitation
The fresh semen of reason is fertilized, and is transferred in IVC droplet after 4h fertilization, by observing cleavage rates after 72h.
Interpretation of result
All test datas are analyzed with SPSS19.0 software, ROS and GSH fluorogram is divided by Image J software
Analysis, pcr gene mrna expression amount are analyzed by lane software, the otherness of variance analysis multiple comparative test data, P<
0.05 indicates significant difference, P<0.01 indicates that difference is extremely significant.Test data with(mean±standard
Deviation it) is indicated with percentage.
After various concentration crocin is added to and cultivates 44h in oocyte in vitro maturation culture solution, maturing rate is observed
(regarding first polar body discharge as maturation), crocin concentration is respectively 0,300ug/ml, 400ug/ml and 500ug/ml.
The results are shown in Table 2, it has been found that addition various concentration crocin (0,300ug/ml, 400ug/ml and
Oocyte in vitro maturation rate (first polar body discharge) 500ug/ml) has extremely significant difference (P<0.01).Maturing rate is respectively
72.80%, 80.04%, 86.80% and 83.20%, maturing rate highest when adding 400ug/ml concentration.
Table 2. adds crocin first polar body discharge rate (nuclear maturation) during oocyte in vitro maturation
Note：The female different expression extremely significant (P of difference of same column shoulder marking-up<0.01).
After various concentration crocin is added to and cultivates 44h in oocyte in vitro maturation culture solution, choose mature MII
Phase egg mother cell detects ROS and GSH concentration.
As a result as shown in Figure 1, addition various concentration crocin (0,300ug/ml, 400ug/ml and 500ug/ml) is right
The discovery of mature oocyte ROS and GSH Concentration Testing, concentration reduces the extremely significant (P of difference to processing group ROS compared with the control group<
0.01), wherein the ROS concentration of addition crocin 500ug/ml is minimum (Figure 1), processing group GSH adds compared with the control group
Add the GSH concentration difference of crocin 300ug/ml not significant, the extremely significant (P of GSH concentration difference of 400ug/ml and 500ug/ml<
0.01), wherein addition 400ug/ml when GSH concentration highest.
After various concentration crocin is added to and cultivates 44h in oocyte in vitro maturation culture solution, choose mature MII
Phase egg mother cell detects SOD, the expression quantity of CAT and GPx gene mRNA.As a result as shown in Figure 2.
In test 3, detection addition various concentration crocin (0,300ug/ml, 400ug/ml and 500ug/ml) is right
The influence of the mrna expression amount of mature oocyte tri- genes of SOD, CAT and GPx.We have found that in sod gene, addition
The 300ug/ml time difference heteropolar significant (P<0.01) without significant difference when, adding 400ug/ml and 500ug/ml.In CAT gene,
Significant difference (P when adding 300ug/ml and 400ug/ml<0.05) without significant difference when, adding 500ug/ml.In GPx gene
Three groups of processing groups have extremely significant difference (P<0.01).
After various concentration crocin is added to and cultivates 44h in oocyte in vitro maturation culture solution, choose mature MII
Phase egg mother cell carries out in vitro fertilization, detection cleavage rates with fresh semen.The results are shown in Table 3
The subsequent spilting of an egg rate in vitro fertilization of egg mother cell of addition hiding crocin during 3. maturation in vitro of table
Note：The female different expression extremely significant (P of difference of same column shoulder marking-up<0.01).
Test 4 in, addition various concentration crocin (0,300ug/ml, 400ug/ml and 500ug/ml) detection at
The cleavage rates of ripe egg mother cell and fresh semen after fertilization add 400ug/ml without significant difference when 300ug/ml is added in discovery
(62.50%) and 500ug/ml (60.83%) time difference heteropolar significant (P<0.01), cleavage rates are up to 62.50% (Table
3)。
Finally it should be noted that：The foregoing is only a preferred embodiment of the present invention, is not intended to restrict the invention,
Although the present invention is described in detail referring to the foregoing embodiments, for those skilled in the art, still may be used
To modify the technical solutions described in the foregoing embodiments or equivalent replacement of some of the technical features.
All within the spirits and principles of the present invention, any modification, equivalent replacement, improvement and so on should be included in of the invention
Within protection scope.
Claims (3)
1. crocin is improving the application in porcine oocytes oxidation resistance and cleavage rates.
2. application as described in claim 1, which is characterized in that the dosage of crocin is 300~500ug/mL.
3. application as described in claim 1, which is characterized in that the dosage of crocin is 400ug/mL.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117158414A (en) * | 2023-11-02 | 2023-12-05 | 黑龙江八一农垦大学 | Pig sperm cryopreservation diluent containing safflower polysaccharide and application thereof |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011142832A2 (en) * | 2010-05-13 | 2011-11-17 | Whitehead Institute For Biomedical Research | Stem cells derived under low oxygen conditions |
| CN103168970A (en) * | 2013-04-17 | 2013-06-26 | 广东通威饲料有限公司 | Compound feed capable of enhancing reproductive capacity of parent dace |
| US20150064287A1 (en) * | 2013-08-27 | 2015-03-05 | National Taiwan Normal University | Method for treating abnormal polyglutamine-mediated diseases |
| CN106659698A (en) * | 2014-09-19 | 2017-05-10 | 霍图斯诺伟司有限责任公司 | Compositions based on saffron for the prevention and/or treatment of corneal dystrophies. |
| CN108823157A (en) * | 2018-07-05 | 2018-11-16 | 浙江大学 | Crocin II and albiflorin are promoting Marrow Mesenchymal Stem Cells In Vitro proliferation and are inhibiting the application in replicative senescence |
| US20180362570A1 (en) * | 2017-06-19 | 2018-12-20 | Gangadhara Ganapati | Nicotinamide riboside derivatives and their uses |
| CN110484494A (en) * | 2019-09-27 | 2019-11-22 | 延边大学 | A method of improving Porcine In Vitro Maturation Oocytes quality |
-
2018
- 2018-06-13 CN CN201810609340.2A patent/CN108841781A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011142832A2 (en) * | 2010-05-13 | 2011-11-17 | Whitehead Institute For Biomedical Research | Stem cells derived under low oxygen conditions |
| CN103168970A (en) * | 2013-04-17 | 2013-06-26 | 广东通威饲料有限公司 | Compound feed capable of enhancing reproductive capacity of parent dace |
| US20150064287A1 (en) * | 2013-08-27 | 2015-03-05 | National Taiwan Normal University | Method for treating abnormal polyglutamine-mediated diseases |
| CN106659698A (en) * | 2014-09-19 | 2017-05-10 | 霍图斯诺伟司有限责任公司 | Compositions based on saffron for the prevention and/or treatment of corneal dystrophies. |
| US20180362570A1 (en) * | 2017-06-19 | 2018-12-20 | Gangadhara Ganapati | Nicotinamide riboside derivatives and their uses |
| CN108823157A (en) * | 2018-07-05 | 2018-11-16 | 浙江大学 | Crocin II and albiflorin are promoting Marrow Mesenchymal Stem Cells In Vitro proliferation and are inhibiting the application in replicative senescence |
| CN110484494A (en) * | 2019-09-27 | 2019-11-22 | 延边大学 | A method of improving Porcine In Vitro Maturation Oocytes quality |
Non-Patent Citations (4)
| Title |
|---|
| ELHAM MOKHBER MALEKI等: "Effects of Crocin Supplementation during In Vitro Maturation of Mouse Oocytes on Glutathione Synthesis and Cytoplasmic Maturation", 《INTERNATIONAL JOURNAL OF FERTILITY & STERILITY》 * |
| HYEJI SHIN等: "Effect of Crocin during In Vitro Maturation on Cytoplasmic Maturation and Embryonic Development of Pig Oocytes after Parthenogenesis", 《韩国动物生命学术研讨会论文集：THE 16TH INTERNATIONAL SYMPOSIUM ON DEVELOPMENTAL BIOTECHNOLOGY》 * |
| XUAN CHEN: "Crocin supplementation during oocyte maturation enhances antioxidant defence and subsequent cleavage rate", 《REPRODUCTION IN DOMESTIC ANIMALS》 * |
| 玄彪: "藏红花素对猪卵母细胞成熟过程中抗氧化性和卵裂率的影响", 《中国优秀硕士学位论文全文数据库医药卫生科技辑》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117158414A (en) * | 2023-11-02 | 2023-12-05 | 黑龙江八一农垦大学 | Pig sperm cryopreservation diluent containing safflower polysaccharide and application thereof |
| CN117158414B (en) * | 2023-11-02 | 2024-02-02 | 黑龙江八一农垦大学 | Pig sperm cryopreservation diluent containing safflower polysaccharide and application thereof |
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