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The Expression of P53 in the Spermatogenic Cells Apoptosis and the Impaction of the Inhibitor of P53 to BCL-2/BAX
Author: XieLi
Tutor: HeLiPing
School: Nanhua University
Course: Human Anatomy and Embryology
Keywords: P53 Cryptorchid Spermatogenic cells apoptosis Bcl-2 Bax AG P53
CLC: R697
Type: Master's thesis
Year: 2008
Downloads: 49
Quote: 0
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Abstract
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Objective: To explore the role of P53' dissertation">P53 on the spermatogenic cells apoptosis and the relationship with the Bcl-2/Bax.Methods: Thirty-two male Sprague-Dawley rats(42day-old) were divided into four groups: sham group, cryptorchid+p53 inhibitor (p-fifty three inhibitor-alpha, PFT-α) group, cryptorchid + dissolvent of PFT-αgroup, cryptorchid groups. The model operated by the literature[10]. Unilateral cryptorchidism was surgically induced in the rats under pento- barbital anaesthesia: a midline abdominal incision was made and then the right testis was sutured to the abdominal wall after the gubernaculum was cut to prevent testis descent. The left testis remained in the scrotum and the right testis was displaced from the scrotum to the abdomen. The gubernaculums of the right testes in the sham operation group were cut before replaced into the scrotum. The rats of cryptorchid + PFT-αgroup and cryptorchid + dissolvent group were injected intraperito- neally PFT-αand dissolvent respectively. The rats were sacrificed on the 7th days after operation. The morphology of spermatogenic epithelium in rats of every group was observed with light microscopy. The apoptosis of spermatogenic cells in the unilateral cryptor chidism was evaluated by TUNEL and FCM. The protein expression level of P53, Bcl-2 and Bax in testes were detected by Western blot and immunohistochemical assay accordingly.Results: In addition to sham group, the right testicle of other groups are less weight than the left testicle; the right testis of cryptorchid group and cryptorchid + dissolvent group dissolved weight significantly lighter than other groups; The orchidopexy of cryptorchid + PFT-αgroup were higher significantly than the cryptorchid grou and cryptorchid + dissolvent group; The differences between the cryptorchid group and cryptorchid + dissolvent group was insignificant. Apoptosis index (AI) was the highest in cryptorchid group and the cryptorchid + dissolvent group, was the lowest in the sham group. The AI in the cryptorchid + PFT-αgroup was lower than the cryptorchid + dissolvent group. The differences were significant. Western Blot analysis showed the protein expression level of P53 and Bax in the testes of the cryptorchid group and the cryptorchid + dissolvent group were much than the other two groups, the sham operation group was the lowest. And the protein expression level of Bcl-2 in the testes of the cryptorchid group and the cryptorchid + dissolvent group is lower than the other two groups, the sham operation group was the hightest. The differences were significant.Conclusion:1.The expression of p53 was increased in the process of germ cell apoptosis by the experimental cryptorchidism.2.PFT-αcould inhibit the germ cell apoptosis caused by the experimental cryptorchidism through increasing the expression of Bcl-2 and decreasing the expression of Bax. Objective: Through that observe aminoguanidine influenced the expression of P53 in the spermatogenic cells apoptosis of cryptorchidism , explore the relationship between NO and P53 in the process of spermatogenic cells apoptosis .Methods: Thirty-two male Sprague-Dawley rats(42day-old) were divided into four groups: sham group, cryptorchid+iNOS inhibitor (Aminoguanidine ,AG) group, cryptorchid + dissolvent of AG(NS) group, cryptorchid groups. The model operated by the literature[10]. Unilateral cryptorchidism was surgically induced in the rats under pento- barbital anaesthesia: a midline abdominal incision was made and then the right testis was sutured to the abdominal wall after the gubernaculum was cut to prevent testis descent. The left testis remained in the scrotum and the right testis was displaced from the scrotum to the abdomen. The gubernaculums of the right testes in the sham operation group were cut before replaced into the scrotum. The rats of cryptorchid + AG group and cryptorchid + NS group were injected intraperito- neally AG and NS respectively. The rats were sacrificed on the 7th days after operation. NO content in the right testis was measured by Greiss method. The activity of NOS in the right testis was measured by colorimetric method. The morphology of spermatogenic epithelium in rats of every group was observed with light microscopy. The apoptosis of spermatogenic cells in the unilateral cryptorchidism was evaluated by TUNEL and FCM. The protein expression level of P53 and iNOS in testes were detected by Western blot and immunohistochemical assay accordingly.Results: In addition to sham group, the right testicle of other groups are less weight than the left testicle; the right testis of cryptorchid group and dissolvent group dissolved weight significantly lighter than other groups; The orchidopexy of cryptorchid + AG group were higher significantly than the cryptorchid grou and the cryptorchid + NS group; The differences between the cryptorchid group and the cryptorchid + NS group was insignificant. Administration of AG could decrease NO content, the activity of NOS and spermatogenic cells apoptosis of the cryptorchid testes, and it could significantly inhibit weight lose of the cryptorchid. Apoptosis index (AI) was the highest in cryptorchid group and the cryptorchid + NS group, was the lowest in the sham group. The AI in the cryptorchid + AG group was lower than the cryptorchid + NS group. The differences were significant. Western Blot analysis showed the protein expression level of P53 in the testes of the cryptorchid group and the cryptorchid + NS group were much than the other two groups, the sham operation group was the lowest. The differences were significant. And the protein expression level of iNOS was lowest in the sham operation group: there were insignificant differences between other three groups.Conclusion:1. AG that reduced the expression of P53 in the experimental cryptorchidism, inhibit the spermatogenic cells apoptosis caused by the experimental cryptorchidism;2. The production of NO increase in the experiment cryptorchidism. It would induce increasing of P53. That result in decreasing the Bcl-2 and increasing the Bax.
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CLC: > Medicine, health > Surgery > Urology ( urinary and reproductive system diseases) > Male genital diseases
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