Effects Of Adiponectin On Functional Development Of Gonads Of Wannanhua Pigs

Wannanhua pigs were superordinary local pigs of Anhui province. The production traits and genetic qualities of Wannanhua pigs were special in Chinese local pigs. Adiponectin, as one of the most abundant adipokines secreted by adipose tissues, had closely relationship with the reproduction particularly. The development patterns of sex hormone in blood, adiponectin receptor mRNA and synthesis-related factors mRNA expressions in gonads were investigated, and their correlation were analyzed in order to approach the inner relationship of adiponectin and reproducion in Wannanhua pigs. In addition, the effects of recombinate adiponectin（rAdp） on cultured leydigs cells and granulose cells of pigs were also studied in this paper in order to study the influence of adiponectin on gonadal hormone synthesis in Wannanhua Pig, and its possible mechanism.1Developmental patterns of serum sex hormone parameters in blood and temporal expression of related genes in Wannanhua pigs’ gonads.The test animals were sampled on0,30,45,90, and180days of age, including10pigs at each different days of age（half males and half females）. The serum concentration of testosterone （T）and estradiol （E₂） in Wannanhua boars and gilts were tested respectively by ELISA. And the expression of AdpR1, AdpR2, LHR, CYP11A1, StAR and CYP19mRNA in gonads were detected by real-time fluorescence quantitative RT-PCR. The results showed as follows:（1） The concentration of serum T in Wannanhua boars decreased during0⁴5d and then kept stabilization, showed significant development patterns as a whole（P<0.01）. The serum E₂concentration in Wannanhua gilts get the peaks at0and45d, and had no difference in30d,90d or180d. The significant development patterns of serum E₂were also seemed in gilts with decreasing at first, then increasing, and the sustaining at last.（2） In testis, there was a significant development pattern of AdpR1mRNA expression, while the expression of AdpR2mRNA just showed a increasing tendency. In ovaries, both AdpR1and AdpR2mRNA expression had a significant developmental patterns（P<0.01or P<0.05）. The expression AdpR1mRNA were higher than AdpR2mRNA in both testis and ovaries.（3）The expression of CYP11A1、 StAR and LHR mRNA had a significant developmental patterns in testis（P<0.01or P<0.05）. The expression of FSHR mRNA but not CYP19mRNAhad a significant developmental pattern in ovaries. （4） The analysis of correlation:In testis, significant positive correlation was found between mRNA expressions of AdpR1and CYP11A1（r=0.587, P<0.05）. The mRNA expression of LHR were significantly correlated with those in CYP11A1（r=0.528, P<0.05） and StAR （r=0.552, P<0.05） respectively. And significant positive correlation was found between CYP11A1and StAR mRNA expression （r=0.709, P<0.01）.In ovaries, the mRNA expression of AdpR1were positively correlated with AdpR2significantly （r=0.380，P<0.05） while the mRNA expression of AdpR1were negatively correlated with CYP19significantly （r=-0.472，P<0.05）.The high expression of AdpR1mRNA in tesis suggested that the action of adiponectin on testis may mediated by AdpR1. The positive correlation between AdpR1and CYP11A1mRNA expression indicated that adiponectin contribute the generation of testosterone.In ovaries, the synthesis of E₂were regulated by both AdpR1and P450arom, which had negative correlation in their mRNA expression. The positive correlation between AdpR1and AdpR2mRNA, and the high expression of AdpR1mRNA indicated that adiponectin decreased the synthesis of E₂mediated byAdpR1.2Effect of adiponectin on sex hormone synthesis in cultured leydigs cells and granulose cells of pigs and its possible mechanismLeydigs cells were isolated abactcrially from testis at10d of Wannanhua boar. Granulose cells were cultured after some treatment. Cells treated with rAdp two days after subculture. ELISA methods were used to detect the concentration of T and E₂, and RT-PCR methods were used to detect the expression of AdpR1、AdpR2、LHR、FSHR、StAR、 CYP11A1、CYP19mRNA in cells after12h treatment. The results showed as follows:（1）The variation of T and E₂concentration in culture solution:T concentration increased significantly after the treatment of2μg/mL rAdp, while T concentration has no significant difference with Compound C+2μg/mL Adp treatment in leydigs cells culture solution. E₂concentration has no significant change after the treatment of2μg/mL rAdp.（2） The variation of mRNA expression of some genes in cells:In leydigs cells, rAdp increased the expression of AdpR1mRNA, but had no significant effects on AdpR2mRNAexpression. rAdp decreased the expression of StAR but had no effects on CYP11A1mRNA. In addition, rAdp increased the expression of AMPK mRNA in leydigs cells. The discrepant results between T concentration in culture solution and mRNA expression of T synthesis-related genes in cells indicated that T concentration may affected by switching some negative feedback mechanism. The increasing of T concentration suggested that the effects of adiponectin on leydigs cells may mediated by AdpR1, activatedAMPK pathway, increased the synthesis of T.In granulose cells, rAdp reduced the expression of AdpR2, CYP19and AMPK mRNA and but did not affect AdpR1mRNA. These results indicated that the action of adiponectin on granulose cells may also mediated by AdpR2, inhibit AMPK pathway, reduce the synthesis of E₂. The results of expreiment in vivo and in vitro suggested that the action of Adp on reproduction of gilts may mediated by both AdpR1and AdpR2.