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1.
将90只大鼠随机分为5组,分别注射10μg.100μL-1 pcISI(T1)、50μg.100μL-1 pcISI(T2)、100μg.100μL-1(T3)pcISI、100μg空载体pcMV-S(V)和100μL生理盐水(S),以探讨在不使用免疫佐剂的情况下,不同剂量的双拷贝抑制素pcISI基因免疫对大鼠卵泡发育、产仔和生殖激素的影响。结果表明,加强免疫能提高抗体P/N值,加强免疫后各剂量组平均抗体P/N值大于2,T3组抗体P/N值显著高于T1和T2组(P<0.05)。T3组成熟卵泡数显著高于对照组(P<0.05),且抗体阳性鼠成熟卵泡数比阴性鼠多12.45个(P<0.05)。除了T1组外,其他2个剂量组产仔数和胎盘数均高于对照组(P<0.05)。抗体阳性鼠胎盘数比阴性鼠多5.09个(P<0.05),产仔数比阴性鼠多5.39个(P<0.05)。抑制素pcISI基因免疫大鼠后促卵泡素(FSH)、雌二醇(E2)和孕酮(P4)含量均高于对照组,T3组的FSH含量与对照组差异显著(P<0.05),T3组的E2含量与对照组差异极显著(P<0.01),各剂量组P4含量与对照组差异均不显著(P>0.05)。这些结果说明,在没有免疫佐剂的前提下,双拷贝抑制素pcISI基因免疫可产生抗体,促进FSH分泌和卵泡发育,增加产仔数。本试验条件下100μg.100μL-1是最佳免疫剂量。  相似文献   

2.
抑制素基因免疫大鼠的免疫应答与基因表达   总被引:1,自引:1,他引:1  
为了研究抑制素基因免疫对大鼠免疫应答和发情的影响及免疫后抑制素的组织分布,将60只大鼠分为5组,每只分别肌肉注射10(T1)、50(T2)、100μg pCIS(T3),50μg pcDNA3.1(V)和生理盐水(S)。ELISA检测抑制素抗体水平,阴道涂片法检测大鼠的发情状况,免疫组化分析抑制素的组织分布。结果显示,不同剂量抑制素基因2次免疫10 d后抗体P/N值均显著升高(P〈0.05),3次免疫10 d后T2和T3组抗体P/N值进一步显著升高(P〈0.05),T3组抗体水平有高于T1和T2组的趋势(P〉0.05);抑制素基因免疫对大鼠的发情无显著影响;3次免疫2周后心脏、肝脏、接种肌肉部位均未检出抑制素;卵巢、肾脏和垂体部位均检出抑制素;而脾脏部位,抑制素质粒免疫组检出抑制素,对照组则未检出抑制素。这些结果表明,免疫剂量和次数的增加没有导致大鼠产生明显的抑制素免疫耐受,抑制素基因免疫大鼠是相对安全的,抑制素的组织分布结果亦为抑制素基因免疫的作用机理研究提供了理论依据。  相似文献   

3.
选择32只60日龄雌性性成熟大鼠分为4组,分别肌肉注射pcDNA-DPPISS-DINH(试验组1)50μg,poD—NA—DPPISS-DINH—mC3d3(试验组2)50μg,空载体pcDNA3.1(对照组1)50μg,生理盐水(对照组2)1mL。首次免疫20、40d后各组分别同剂量同样免疫程序加强免疫注射1次。结果发现,试验1和2组卵巢的重量都显著高于对照组1和2,但试验1和2组之间的差异不显著(P〉0.05);试验2组卵巢成熟卵泡发育个数和FSH的浓度显著高于试验1组和对照1和2组(P〈0.05);加强免疫第40天,试验1、2组的E1的浓度分别显著高于本试验组免疫前第0天,免疫后第20天的激素浓度;试验组与对照组在各个免疫试验时间LH的浓度在统计上差并不显著(P〉0.05)。结果证明,应用重组抑制素真核表达质粒pcDNA-DPPISS-DINH—mC3d3(试验组2)免疫动物可以促进大鼠卵泡的发育,可提高血浆FSH、E2水平,为抑制素基因免疫大动物提供了试验依据。  相似文献   

4.
90只大鼠随机分为5组(n=18),分别肌肉注射10、50、100μg抑制素与乙肝表面抗原融合基因表达质粒(pCIS)、50μg空载体(pcDNA3.1)和100μL生理盐水。20 d后加强免疫1次,对每组中的12只大鼠进行2次加强免疫。结果发现抑制素抗体P/N值随着免疫次数的增加而提高。100μg剂量组2次和3次免疫的成熟卵泡发育数分别比对照组多6.9和7.5个(P<0.05)。3次免疫后大鼠成熟卵泡发育数比2次免疫后显著提高(35.2±2.73 vs 31.0±0.92,P<0.05)。抑制素基因免疫组的胎盘数和窝产仔数高于对照组(P>0.05),抗体阳性鼠高于阴性鼠(P<0.05)。pCIS 3次免疫后抗体阳性鼠的抗体水平与成熟卵泡发育数的相关系数为0.45(P>0.05),与胎盘数的相关系数为0.77(P<0.05)。pCIS免疫大鼠动情期和产后血浆FSH水平高于对照组,抗体阳性鼠的血浆FSH水平高于阴性鼠,其中2次免疫后阳性组动情期FSH浓度极显著高于阴性组(P<0.01)。这些结果表明,抑制素基因免疫大鼠可促进大鼠的卵泡发育,提高血浆FSH水平,为抑制素基因免疫大动物提供了试验依据。  相似文献   

5.
抑制素基因免疫对小鼠生殖的影响   总被引:32,自引:5,他引:27  
用编码抑制素α(1-32)的基因与pcDNA3.1真核表达质粒进行重组,构建抑制素基因疫苗NH。将30只2周龄ICR小鼠随机分为3组(每组10只),试验Ⅰ,Ⅱ组分别经脂质体介导肌肉注射iINH15μg()0μL,25μg(100μL),对照组注射免疫空载体pcDNA3.115μg(100μL)。首次免疫后间隔3周加强免疫1次。小经pINH基因免疫后,用ELISA可从血浆中检测到抑制素抗体,表明pINH可在活体肌细胞内表达,表达产物具有制素抗原性,能激活免疫系统产生抑制素抗体。抑制素抗体P/N>2视为阳性,阳性率为30%(6/20)。抑制素抗体阳性小鼠与阴性小鼠的产仔数,初生重和窝重等没有显著差异(P>0.05)。抗体阳性组首次免疫后,雌鼠血浆FSH浓度略有升高,经过加强免疫后显著升高(P<0.05)。由此可以看出,抑制素基因免疫可以诱导产生抗抑制素抗体,并促进促卵泡素的分泌。  相似文献   

6.
抑制素pCIS基因免疫对黄牛卵泡发育和生殖激素的影响   总被引:2,自引:1,他引:1  
分别用O(C)、0.75(T1)、1.50(T2)、2.25(T3)和3.0(T4)nag·头'-1的抑制素pCIS基因疫苗免疫58头黄牛,以探讨抑制素pCIS基因免疫对卵泡发育和生殖激素的影响.结果表明,抑制素pCIS基因免疫组的大卵泡(d≥10 mm)数显著高于对照组(P<0.01),中卵泡(7 mm≤d<10 mm)数和小卵泡(4 mm≤d<7 mm)数与对照组均没有显著差异(P>0.05),T1组左右两侧成熟卵泡大小与对照组差异不显著(P>0.05),其它3组两侧成熟卵泡均显著大于对照组(P<0.05).抑制素pCIS基因免疫黄牛的促卵泡素(FSH)平均含量高于对照组,且在加强免疫阶段差异显著(P<0.05);加强免疫后雌二醇(E2)和孕酮(P4)含量均显著增加,且均与对照组差异显著(P<0.05).这些结果表明,抑制素pCIS基因免疫可促进FSH分泌,进而影响黄牛卵泡发育和成熟.  相似文献   

7.
选择32只60日龄雌性性成熟大鼠分为4组,分别肌肉注射pcDNA-DPPISS-DINH(试验组1) 50 μg,pcDNA-DPPISS-DINH-mC3d3 (试验组2) 50 μg,空载体pcDNA3.1 (对照组1) 50 μg,生理盐水(对照组2)1 mL.首次免疫20、40 d后各组分别同剂量同样免疫程序加强免疫注射1次.结果发现,试验1和2组卵巢的重量都显著高于对照组1和2,但试验1和2组之间的差异不显著(P>0.05);试验2组卵巢成熟卵泡发育个数和FSH的浓度显著高于试验1组和对照1和2组(P<0.05);加强免疫第40天,试验1、2组的E2的浓度分别显著高于本试验组免疫前第0天,免疫后第20天的激素浓度;试验组与对照组在各个免疫试验时间LH的浓度在统计上差异不显著(P>0.05).结果证明,应用重组抑制素真核表达质粒pcDNA-DPPISS-DINH-mC3d3(试验组2)免疫动物可以促进大鼠卵泡的发育,可提高血浆FSH、E2水平,为抑制素基因免疫大动物提供了试验依据.  相似文献   

8.
抑制素基因免疫对京白鸡产蛋性能的影响   总被引:2,自引:0,他引:2  
120只380 d京白鸡随机分为4组,分别腿部肌肉注射0、50、100、150 μg抑制素真核表达质粒pcISI,20 d后加强免疫1次.ELISA检测抗抑制素抗体水平,观察产蛋量,并进行鸡蛋品质测定.免疫4个月后全部捕杀,统计各级卵泡数.结果表明,抑制素基因加强免疫后抗体水平显著升高(P<0.05),3个剂量组间差异不显著(P>0.05).试验组全期产蛋量极显著高于对照组(P<0.01),大白泡和小白泡比对照组显著增多(P<0.05),优势卵泡、小黄泡和鸡蛋品质与对照组差异不显著(P>0.05).表明抑制素基因免疫京白鸡可产生抗抑制素抗体,并能提高京白鸡的产蛋性能.  相似文献   

9.
选择40头南阳黄牛,用3.0毫克抑制素基因重组质粒(PCIS,免疫组,n=30)或同剂量的生理盐水(对照组,n=10)间隔21天注射2次,分别在首次免疫当天、首次免疫后第10天和第21天、加强免疫后第10天和第45天收集血清,应用放射免疫法(RIA)测定处理后不同时期血清中激素水平。结果表明:在5个采血时间点内.加强免疫后第10天抗体阳性牛比率最高.为37.5%,与其他采血时间点有显著差异。免疫组促卵泡素(FSH)平均含量比对照组高,但差异不显著;17-β雌二醇(E2)和孕酮(P4)水平在加强免疫后2组有显著性差异。  相似文献   

10.
抑制素基因免疫的免疫反应性   总被引:9,自引:1,他引:9  
通过应用抑制素真核表达质粒pcINH和抑制素融合表达质粒pCIS分别经肌肉注射免疫大鼠的2个系列试验,检测了免疫鼠抑制素抗体水平及抗体阳性鼠的比例。结果,抑制素质粒pcINH经脂质体介导免疫大鼠,50%(13/26)的个体产生了抑制素阳性抗体。40μg组产生的抗体水平最高;抑制素融合表达质粒pCIS免疫经盐酸普鲁卡因处理的大鼠,2次免疫获得了38.9%(23/54)的抗体阳性率。3次免疫获得55.6%(20/36)的抗体阳性率,100μg组产生的抗体水平最高;加强免疫可提高抗体的水平。但免疫剂量的增加并不一定能增加抗体阳性鼠的比例。  相似文献   

11.
本研究采用新疆细毛羊抑制素成熟区序列重组融合蛋白(roINH)作为免疫原,以Montanide (MON)和Freund's (FA)作为佐剂,主动免疫SD大鼠,测定生殖激素及卵巢发育情况,评价佐剂效应.选用72只9~10周龄性成熟雌性SD大鼠,随机分为3组,分别皮下注射生理盐水(对照组)、roINH+ MON(MON组)和roINH+FA (FA组).各组大鼠每20 d免疫1次,连续免疫3次.结果,MON组产生的抗体滴度显著高于FA组(P<0.05);与对照组相比,注射roINH+MON和roINH+FA均可极显著提高大鼠血清FSH平均含量(P<0.01),显著提高P峰值(P<0.05),而FSH及P平均含量及峰值在2个免疫组间无显著差异(P>0.05);LH含量和峰值在2个试验组间无显著差异(P>0.05); MON和FA组成熟卵泡数无显著差异(P>0.05),但均显著高于对照组(P<0.05);MON组脓包直径及炎症反应小于FA组.结果表明,应用roINH作为免疫原并配以MON或FA佐剂免疫大鼠均能取得较好的免疫效果,且MON佐剂炎症反应小于FA佐剂.  相似文献   

12.
The hypothesis of the present study is that active immunization of cows against inhibin would neutralize endogenous inhibin, increase circulating levels of follicle stimulating hormone, and subsequently affect follicular dynamics and the ovulation rate during the estrous cycle. Thirteen cows were immunized against inhibin alpha-subunit and, 6 cows were immunized with a placebo. Both groups were given 4 booster immunizations 7, 14, 21, and 34 weeks after the primary injection. Ovaries were examined daily after the 2nd, 3rd, and 4th booster immunizations by transrectal ultrasonography for 25 days. After the 4th booster immunization, blood samples were collected daily for one complete estrous cycle to measure FSH and LH. The results showed that the immunized cows generated antibodies against inhibin, and that they had higher FSH levels compared with the controls. The number of follicular waves during the estrous cycle was higher in the immunized cows (3 or 4 waves) than in the controls (2 or 3 waves). Moreover, the immunized cows had a greater number of follicles during the estrous cycle compared with the control cows. The maximum number of follicles was 14.8 +/- 1.7 vs 5.4 +/- 0.2 in inhibin-immunized and control cows, respectively, during the first follicular wave and 13.9 +/- 1.9 vs 5.6 +/- 0.7, respectively, during the ovulatory wave. Multiple ovulations were increased in the immunized cows. However, the ovulation rate varied greatly in the immunized animals. In conclusion, immunization against inhibin increased FSH secretions during the estrous cycle in the cows. Moreover, the immunized cows had a greater number of follicular waves during the estrous cycle and a greater number of follicles, and this could be used as a potential source of oocytes for use in IVF/embryo transfer programs.  相似文献   

13.
Eight heifers, aged 16–17 months and showing normal oestrous cycles, were immunized against a recombinant porcine inhibin α subunit immunogen, together with another 10 heifers of the same age as controls and treated with placebo immunogen. Primary (1 mg immunogen) and two booster (0.5 mg immunogen each) immunizations were administered at 28‐day intervals. Ten days after the second booster immunization, both groups of heifers underwent a superovulation treatment. Each animal was given an intravaginal progesterone releasing sponge, which was withdrawn 7 days following an i.m. injection of 0.5 mg cloprostenol. Heifers were treated with FSH for 4 days and artificially inseminated after oestrus occurred. The embryos were flushed and evaluated 7 days after insemination. Immunization significantly (p < 0.01) increased blood antibody titres against recombinant porcine inhibin α subunit, from pre‐immunizaion and control values of approximately 0.06 of ELISA 450 nm reading to 0.6 to 0.7 after two or three immunizations. The immunized heifers produced on average 15.8 ± 2.8 embryos, significantly (p < 0.05) higher than the yield of 8.3 ± 1.5 in the controls. The number of transferable embryos were non‐significantly higher in immunized than in control heifers (9.6 ± 3.1 vs 5.8 ± 1.6, p > 0.05). The peak plasma oestradiol concentrations were significantly higher in immunized than in control heifers, both immediately after FSH treatment and 20 days thereafter. Plasma P4 concentrations after superovulation were in the range of 20 ng / ml in the immunized heifers, significantly (p < 0.05) higher than the values approximately 15 ng / ml in control heifers. These results indicated that prior immunization against inhibin α subunit stimulated production of antibodies against inhibin, which enhanced follicular developmental response to superovulation and lead to higher yield of total and transferable embryos. Therefore immunization combined with the conventional superovulatory gonadotrophin treatment, can be a simple and efficient method to produce low cost bovine embryos.  相似文献   

14.
In the present study, two experiments were conducted to induce superovulation in goats using passive and active immunization against inhibin. In the first experiment, two groups of goats were given an intravenous injection of either 10 ml normal goat serum (control; n=6) or inhibin antiserum developed against [Tyro30]-inhibin alpha (1-30) (passively immunized; n=6) 48 h before treatment with PGF2alpha. In the second experiment, two groups of goats were immunized with inhibin vaccine (actively immunized; n=5) or Freund's adjuvant (control; n=5) followed by three booster immunizations at 4 week intervals. Blood samples were collected for determination of FSH, LH, estradiol-17beta, and progesterone. Ultrasonography was used to determine ovarian activity at PGF2alpha injection and ovulation rate one week after estrus. In both experiments, there was a significant increase in plasma FSH concentration compared with the controls. However, the pattern of the FSH levels was different between the passively and actively immunized goats. The numbers of follicles in passively and actively immunized goats (22.4 +/- 2.3 and 18.6 +/- 2.1, respectively) were significantly greater than those in the controls (2.6 +/- 0.4 and 2.3 +/- 0.4, respectively). In addition, the ovulation rate was greater in the immunized animals compared with the controls. Therefore, either passive or active immunization against inhibin could be used to induce superovulation in goats.  相似文献   

15.
Two pony mares were immunized against recombinant porcine inhibin alpha subunit three times with 39 day intervals. Clinical findings and endocrinological changes before immunization were taken as the control. The first significant rise in the anti-inhibin titre (P<0.05) in the circulation was found 27 days after the first injection. Maximum binding activity was reached by the 12th day after the second booster dose. The number of small, medium and large sized follicles had increased significantly compared to before immunization (11.75 +/- 4.30, 2.75 +/- 0.69 and 2.51 +/- 0.63 vs 6.50 +/- 1.43, 1.83 +/- 0.44 and 1.33 +/- 0.38, respectively), but the ovulation rate remained unchanged after immunization. The average plasma concentration of FSH and estradiol-17beta during the estrous cycle increased significantly (P<0.05) after immunization. These results suggest that immunization against inhibin is a useful tool to increase the number of ovarian follicles during the estrous cycle of pony mares. Moreover, the present study supported the concept that inhibin plays a major role in the control of follicular growth through its inhibitory effect on FSH secretion synergistically with steroid hormones.  相似文献   

16.
Inhibin is a gonadal hormone that inhibits the release of follicle stimulating hormone (FSH) from the anterior pituitary gland. The objective of this study was to determine whether active immunization of male and female rats against inhibin rich, steroid-free bovine follicular fluid would increase inhibin antibody titre, onset of female puberty, pregnancy rate, litter size, testis weights, testosterone concentration and serum FSH. Immunization of rats with steroid free bovine follicular fluid stimulated production of anti-inhibin antibodies that immunoneutralized endogenous inhibins and increased levels of circulating FSH in immunized males. Inhibin immunoneutralization resulted in early vaginal opening in immunized females compared with controls and pregnancy rates were increased when immunized female rats were mated with immunized males. However, serum testosterone, testis weights and potential litter size remained unchanged. We conclude that methods to immunoneutralize inhibin may have merit as therapeutic procedures to enhance reproductive performance in domestic animals.  相似文献   

17.
The gonadal hormone inhibin regulates daily sperm production (DSP) indirectly through negative feedback control of FSH secretion and may also affect DSP via direct actions within the testis. Studies attempting to increase DSP through the immunization against inhibin have yielded equivocal results. The current study compared 2 inhibin antigens for effects on DSP and hormone secretion. Hampshire ram lambs (BW = 42 +/- 2 kg; age = 113 +/- 3 d) were assigned randomly to 3 groups: 1) control (n = 4); 2) alpha-peptide conjugate (PTC, n = 6); and 3) alpha-subunit (SUB, n = 6). Antigen PTC consisted of an alpha-inhibin, N-terminal, 25-amino acid peptide conjugated to ovalbumin. Antigen SUB was the complete inhibin alpha-subunit. Lambs were immunized on d 0 (June 19, 2006), 18, 38, and 63. Body weight was recorded on immunization days and scrotal circumference on d 63. Blood samples were collected on d 0, 7, 14, 18, 28, 35, 38, 49, 56, 63, and 70. Rams were slaughtered on d 71. Testes were weighed, and parenchyma was obtained for DSP determination. Plasma alpha-inhibin antibody titer and LH, FSH, and testosterone concentrations were measured. alpha-Inhibin antibody titer was first detectable on d 14 in both PTC- and SUB-immunized ram lambs and generally increased thereafter. Mean DSP per gram of testis (DSP/g) was increased (P < 0.01) 26% in PTC- and SUB-immunized ram lambs over that in control ram lambs. Total DSP per ram lamb and testes weight did not differ among the 3 treatment groups. Variation in DSP per ram lamb and testes weight were greater (P = 0.05) in PTC- and SUB-immunized ram lambs than in control ram lambs. Plasma FSH concentrations were similar in PTC- and SUB-immunized ram lambs. Immunization against either alpha-inhibin antigen did not alter LH, testosterone, BW, or scrotal circumference. Findings indicate that 1) the 2 alpha-inhibin antigens increase DSP/g to similar extents; 2) alpha-inhibin antibody may act at least in part through an intratesticular mechanism because DSP/g was increased in some animals without concomitant increases in FSH; and 3) immunization against alpha-inhibin may affect testes weight by actions independent of those that regulate DSP/g.  相似文献   

18.
A synthetic fragment representing the N-terminal 25 amino acid residues of the alpha-subunit of ovine inhibin (alpha-IF) was coupled to human alpha-globulin (h alpha-G) and used as an antigen. In Exp. 1, ovine antiserum generated against alpha-IF-h alpha-G was shown in vitro to neutralize inhibin bioactivity contained in ovine follicular fluid. In Exp. 2, 18 lambs were immunized with .3, .6 and 1.2 mg alpha-IF-h alpha-G or equivalent doses of h alpha-G. Antibody titer to alpha-IF was detected only in serum from lambs immunized against alpha-IF-h alpha-G and was first detected 27 +/- 2 d after primary immunization. Thereafter, antibody titers increased steadily. The degree of antibody responses was unrelated to antigen dose and differed among lambs. Plasma FSH concentrations were unchanged, whereas LH concentrations were lower (P less than .001) in sheep immunized against alpha-IF-h alpha-G. Ovulation rate was increased (3.5 +/- .5 vs 1.5 +/- .1; P less than .01) in lambs immunized against alpha-IF-h alpha-G. Ovulation rate was similar among animals receiving different antigen doses and increased with time after primary immunization (P less than .01). At estrous periods occurring approximately 34, 50, 74 and 107 d after primary immunization, respective ovulation rates were 157, 169, 207 and 450% of control values. Ovulation rate and antibody titer were correlated positively (pooled r = .95; P less than .01) within lambs. In Exp. 3, three lambs were immunized with .25 mg unconjugated alpha-IF; this was nonantigenic. In conclusion, the use of a synthetic fragment of the alpha-subunit of ovine inhibin as a hapten elicits an antibody capable of neutralizing inhibin bioactivity in vitro and increasing ovulation rate in vivo.  相似文献   

19.
旨在探讨管花肉苁蓉醇提物对大鼠血液生化指标及心血管功能的影响,为管花肉苁蓉对心血管功能不全的辅助治疗作用提供现代医学研究佐证。将24只大鼠随机分为低、中、高剂量组和对照组,分别灌胃0.1、0.2、0.3 g/(100 g·BW)的管花肉苁蓉乙醇提取物和同体积的生理盐水,连续给药30 d。试验结束时,测定并比较各给药组与对照组的主要血液生化指标、心血管功能相关酶和激素水平、脏器系数以及颈动脉平滑肌的运动指标。结果表明,低剂量管花肉苁蓉醇提物显著提高大鼠血清ALT含量(P〈0.05),但低、中、高剂量管花肉苁蓉醇提物对其他血液生化指标影响均不显著(P〉0.05);对大鼠3种心肌酶活力的影响不显著(P〉0.05);低、中剂量组的Ang-Ⅱ水平显著低于对照组(P〈0.05),但EPO活力、ALD水平的变化不显著(P〉0.05);对大鼠的增重率、脏器系数影响不显著(P〉0.05),对颈动脉平滑肌运动指标有一定的降低趋势(P〉0.05)。综上提示,总体来看,管花肉苁蓉醇提物对大鼠的血液生化指标及心血管功能有一定的影响。  相似文献   

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