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去甲腎上腺素誘導(dǎo)HSP70的表達(dá)對隱睪生殖細(xì)胞凋亡的影響

發(fā)布時間:2018-05-08 15:03

  本文選題:去甲腎上腺素 + 隱睪 ; 參考:《遵義醫(yī)學(xué)院》2013年碩士論文


【摘要】:目的:通過比較去甲腎上腺素誘導(dǎo)HSP70的表達(dá)對隱睪癥大鼠生殖細(xì)胞凋亡的影響,探討去甲腎上腺素對隱睪癥的治療作用與機(jī)制,為隱睪癥的臨床治療提供理論實驗依據(jù)。 方法:22日齡雄性SPF(specific pathogen-free animal)級SD大鼠96只,體重40-55g,經(jīng)隨機(jī)數(shù)字表處理后隨機(jī)分為3組:空白組A組(n=32)空白對照組,不采取任何干預(yù)措施;剩余兩組建立單側(cè)隱睪模型后分別施予不同干預(yù)措施,隱睪模型組B組(n=32)隱睪模型生理鹽水處理組(按5ml/kg/d劑量腹腔注射給藥);隱睪模型藥物處理(用生理鹽水配成O.1mg/ml,按0.5mg/kg/d劑量腹腔注射給藥)組C組(n=32)。并于建立模型后的當(dāng)天,第5,10,15d按隨機(jī)的方法取出各組SD大鼠8只心臟穿刺采血3ml,作血清抗體分析用,然后斷頸處死SD大鼠迅速取出隱睪側(cè)睪丸稱重后,按常規(guī)方法石蠟包埋固定,標(biāo)記后留做HE染色觀查并檢測曲細(xì)精管生育力指數(shù)(tubular fertility index, TFI)、平均曲細(xì)精管直徑(mean tubular diametar, MTD),免疫組化,凋亡檢測用。 結(jié)果: (1)各組大鼠隱睪側(cè)睪丸重量的變化:睪丸手術(shù)后5d時,B組睪丸重量就明顯下降(P0.05),隨后持續(xù)下降,到15d時睪丸重量僅為A組的35%。C組變化較小,10d開始下降,與A組、B組的差異均有統(tǒng)計學(xué)意義(P0.05)。 (2)各組大鼠血清抗精子抗體(AsAb)IgG、IgA、IgM定量測定:睪丸手術(shù)后5d時,B組睪丸IgG和IgA水平開始升高(P0.05),且隨著時間的延長,升高越明顯;而B組睪丸IgM水平手術(shù)后5d時就明顯上升(P0.05),10d后開始下降。C組變化較小與A組、B組的差異均有統(tǒng)計學(xué)意義(P0.05),IgG和IgA水平手術(shù)后5d時升高不明顯,無統(tǒng)計學(xué)意義(P0.05),10d后開始上升,與A組、B組比較,有統(tǒng)計學(xué)意義(P0.05),與A組、B組比較,有統(tǒng)計學(xué)意義(P0.05),但升高水平較B組低,而IgM水平手術(shù)后5d時就明顯上升(P0.05),后隨時間延長逐漸降低。 (3)各組大鼠組織學(xué)觀察:B、C組MTD與TFI在手術(shù)后5d明顯降低,與A比較組有統(tǒng)計學(xué)意義(P0.05), C組術(shù)后10d后MTD與TFI有所升高,與A組和B組比較,有統(tǒng)計學(xué)意義(P0.05)。 (4)各組大鼠細(xì)胞凋亡水平變化:B組于隱睪模型建立后在5d時就凋亡明顯增加(P0.05),且隨時間的延長逐漸增加,C組隱睪模型建立后在5d時就凋亡明顯增加(P0.05),且隨時間的延長逐漸增加,但無B組明顯,與A組、B組比較,均有統(tǒng)計學(xué)意義(P0.05); (5)各組大鼠Hsp70表達(dá)量變化:在A組睪丸組織中,Hsp70呈中等程度表達(dá)于精母細(xì)胞和精子細(xì)胞的細(xì)胞核內(nèi)。B組Hsp70于隱睪模型建立后在5d時就明顯減少(P0.05),10d時減少達(dá)到高峰。到隱睪術(shù)后15d時,無明顯Hsp70表達(dá);C組Hsp70隱睪模型建立后在5d也明顯減少(P0.05),10d后Hsp70表達(dá)有所增加,與A組、B組比較,均有統(tǒng)計學(xué)意義(P0.05)。 結(jié)論: (1)去甲腎上腺素可抑制大鼠手術(shù)誘導(dǎo)隱睪后生殖細(xì)胞細(xì)胞凋亡; (2)去甲腎上腺素可上調(diào)大鼠手術(shù)誘導(dǎo)隱睪后生殖細(xì)胞HSP70的表達(dá)。
[Abstract]:Objective: to compare the effect of norepinephrine induced HSP70 expression on germ cell apoptosis in rats with cryptorchidism, to explore the therapeutic effect and mechanism of norepinephrine on cryptorchidism, and to provide theoretical and experimental evidence for clinical treatment of cryptorchidism. Methods Ninety-six male SPF(specific pathogen-free animal- grade SD rats at the age of 22 days, weighing 40-55 g, were randomly divided into three groups: blank group A (group A) and control group (n = 32). The other two groups were given different intervention measures after establishing unilateral cryptorchidism model. The rats in the cryptorchidism model group (group B) were treated with normal saline (intraperitoneal injection of 5ml/kg/d) and group C (treated with 0.1 mg / ml of normal saline and intraperitoneal injection of 0.5mg/kg/d). On the day after the establishment of the model, 8 SD rats in each group were randomly collected for 3 ml of blood by heart puncture on the 15th day after the establishment of the model, and then the SD rats were killed by cervical amputation, and the testis of the cryptorchidism side were quickly removed and weighed. The seminiferous tubules were fixed with paraffin wax and then stained with HE staining. The fertility index of seminiferous tubules was detected by tubule fertility index, and the mean diameter of seminiferous tubules was mean tubular diametar, MTDX, immunohistochemistry, apoptosis detection. Results: (1) the changes of testicular weight in the cryptorchidism side of rats in each group: the testis weight of group B significantly decreased at 5 days after testicular operation, and then continued to decrease, and at 15 days the weight of testis in group A was only 35.C, the change of testicular weight was less than that in group A and the change began to decrease at 10 days after testicular surgery. The difference between group A and group B was statistically significant (P 0.05). (2) quantitative determination of serum anti-sperm antibody (Asab) IgG IgG and IgA in rats in each group: the levels of IgG and IgA in testis of group B began to increase at 5 days after testicular operation, and the increase was more obvious with the prolongation of time. However, the level of IgM in testis of group B increased significantly 5 days after operation and began to decrease 10 days after operation. The difference between group C and group A was statistically significant. The levels of IgM and IgA in group B were not significantly increased at the 5th day after operation, but they began to rise after 10 days without statistical significance. Compared with group A and group B, there was significant difference between group A and group B (P 0.05), but the level of elevation was lower than that of group B, but the level of IgM increased significantly at 5 days after operation, and then decreased gradually with the prolongation of time. (3) histological observation showed that MTD and TFI in group C were significantly lower than those in group A on the 5th day after operation, and there were significant differences between group A and group A (P 0.05), MTD and TFI in group C were increased 10 days after operation, compared with group A and group B, there was significant difference between group A and group B (P 0.05). (4) the apoptosis level of rats in group B increased significantly at the 5th day after the establishment of the cryptorchidism model, and gradually increased with the extension of time, and the apoptosis of the cryptorchidism model in group C increased significantly at the 5th day after the establishment of the model, and gradually increased with the extension of time. But there was no significant difference between group B and group A (P 0.05). (5) the changes of Hsp70 expression: in group A, the expression of Hsp70 in spermatocytes and spermatocytes was moderate. Group B showed a significant decrease in the expression of Hsp70 in the nucleus of spermatocytes and spermatozoa at 5 days after the establishment of the cryptorchidism model, and the decrease of Hsp70 reached its peak at 10 days after the establishment of the cryptorchidism model. On the 15th day after operation, the expression of Hsp70 in group C was significantly decreased after the establishment of the model of cryptorchidism without Hsp70 expression. The expression of Hsp70 in group C was significantly lower than that in group A (P 0.05) 10 days after operation, and there was significant difference between group A and group B (P 0.05). Conclusion: 1) norepinephrine could inhibit the germ cell apoptosis induced by operation in rat cryptorchidism. (2) norepinephrine could up-regulate the expression of HSP70 in germ cells of rats after operation induced by cryptorchidism.
【學(xué)位授予單位】:遵義醫(yī)學(xué)院
【學(xué)位級別】:碩士
【學(xué)位授予年份】:2013
【分類號】:R726.9

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