發(fā)布時(shí)間：2018-01-19 22:29

  本文關(guān)鍵詞： α-硫辛酸 卡那霉素 耳蝸 p38絲裂原活化蛋白激酶 c-Jun氨基末端激酶　出處：《遼寧醫(yī)學(xué)院》2012年碩士論文　論文類型：學(xué)位論文

【摘要】：目的 研究α-硫辛酸（alpha-lipoic acid，LA）對(duì)卡那霉素（kanamycin，KM）致毒小鼠耳蝸磷酸化p38絲裂原活化蛋白激酶（phosphorylated p38mitogen-activated protein kinase， p-p38MAPK）和磷酸化c-Jun氨基末端激酶（phosphorylated c-Jun N-terminal kinase，p-JNK）表達(dá)的影響，探討LA對(duì)KM耳毒性損傷的防護(hù)作用及其機(jī)制。 方法 56只健康BALB/c小鼠隨機(jī)分成對(duì)照組、KM組、KM＋LA組和LA組，每組14只。各組動(dòng)物均每天皮下注射2次，連續(xù)給藥14d。應(yīng)用免疫組織化學(xué)SABC法、顯微圖像分析技術(shù)以及蛋白質(zhì)印跡檢測(cè)觀察小鼠耳蝸中p-p38MAPK和p-JNK的表達(dá)；同時(shí)結(jié)合聽腦干反應(yīng)（auditory brainstem response，ABR）測(cè)試，觀察用藥前后小鼠聽力的變化。 結(jié)果 1．連續(xù)用藥14d后，，在各刺激頻率下，KM組小鼠ABR閾移明顯增大，并且與對(duì)照組比較差異顯著（P＜0.01）；KM＋LA組小鼠ABR閾移雖然在用藥后也有增大，但較KM組明顯減小（P＜0.01）。 2．對(duì)照組耳蝸外毛細(xì)胞、螺旋神經(jīng)節(jié)及血管紋中均可見p-p38MAPK和p-JNK表達(dá)，其陽(yáng)性免疫反應(yīng)產(chǎn)物呈棕黃色，分布于胞漿及胞核內(nèi)。KM組、KM＋LA組和LA組小鼠耳蝸中p-p38MAPK和p-JNK陽(yáng)性反應(yīng)產(chǎn)物的分布與對(duì)照組大致相同，但KM組的陽(yáng)性染色比對(duì)照組明顯加深；而KM＋LA組的陽(yáng)性染色則較KM組明顯減弱。顯微圖像分析結(jié)果顯示，KM組小鼠耳蝸p-p38MAPK和p-JNK表達(dá)較對(duì)照組明顯增強(qiáng)（P＜0.01）；而KM＋LA組小鼠耳蝸p-p38MAPK和p-JNK的表達(dá)則明顯弱于KM組（P＜0.01）。 3．蛋白質(zhì)印跡檢測(cè)結(jié)果顯示，對(duì)照組小鼠耳蝸p-p38MAPK和p-JNK的電泳條帶較弱，而KM組小鼠耳蝸p-p38MAPK和p-JNK的電泳條帶均明顯強(qiáng)于對(duì)照組，KM＋LA組小鼠耳蝸p-p38MAPK和p-JNK的電泳條帶則較KM組明顯減弱。半定量分析結(jié)果顯示，KM組小鼠耳蝸p-p38MAPK和p-JNK的蛋白表達(dá)水平均較對(duì)照組明顯增高（P＜0.01）；而KM＋LA組小鼠耳蝸p-p38MAPK/β-actin比值和p-JNK/β-actin比值則均較KM組明顯減小，即p-p38MAPK和p-JNK的蛋白表達(dá)水平表達(dá)較KM組明顯降低。 結(jié)論 p38MAPK和JNK介導(dǎo)了KM對(duì)BALB/c小鼠的耳毒性損傷，LA可通過(guò)顯著抑制KM所致p-p38MAPK和p-JNK的高表達(dá)，從而有效防護(hù)KM的耳毒性，這可能是LA發(fā)揮防護(hù)作用的機(jī)制之一。
[Abstract]:Purpose To study the effect of alpha-lipoic acid lac (偽 -lipoic acid) on kanamycin (kanamycin). KM- induced phosphorylation of p38 mitogen-activated protein kinase (p38) in the cochlea of mice. Phosphorylated p38mitogen-activated protein kinase. P-p38 MAPK) and phosphorylated c-Jun N-terminal kinase. To investigate the protective effect of LA on km ototoxicity and its mechanism. Method 56 healthy BALB/c mice were randomly divided into control group (km + LA) and LA group (14 rats in each group). The expression of p-p38 MAPK and p-JNK in mouse cochlea was observed by immunohistochemical SABC method, microimage analysis and Western blotting. At the same time, the auditory brainstem response (ABR) and auditory brainstem response (ABR) test were used to observe the hearing changes of mice before and after treatment. Results 1.After 14 days of continuous administration, the ABR threshold shift of mice in KM group increased significantly at different stimulation frequencies, and the difference was significant compared with that of control group (P < 0.01); The threshold shift of ABR in KM+LA group increased after administration, but decreased significantly compared with km group (P < 0.01). 2. The expression of p-p38 MAPK and p-JNK was found in the cochlear outer hair cells, spiral ganglion and stria vascularis in the control group. The positive immunoreactive products of p-p38 MAPK and p-JNK were brown and yellow. The distribution of p-p38 MAPK and p-JNK positive products in the cochlea of the cytoplasmic and cytoplasmic. Km + LA group and LA group was approximately the same as that in the control group. But the positive staining of km group was significantly deeper than that of control group. The positive staining in KM+LA group was significantly lower than that in km group. The expression of p-p38 MAPK and p-JNK in km group was significantly higher than that in control group (P < 0.01). The expression of p-p38 MAPK and p-JNK in KM+LA group was significantly weaker than that in km group (P < 0.01). 3. The results of Western blot analysis showed that the electrophoretic bands of p-p38 MAPK and p-JNK were weak in the cochlea of control mice. The electrophoretic bands of p-p38 MAPK and p-JNK in km group were significantly stronger than those in control group. The electrophoretic bands of p-p38 MAPK and p-JNK in cochlea of KM+LA group were significantly lower than those in km group. The expression of p-p38 MAPK and p-JNK protein in km group was significantly higher than that in control group (P < 0.01). However, the ratio of p-p38 MAPK / 尾 -actin and p-JNK/ 尾 -actin in cochlea of KM+LA group was significantly lower than that in km group. The protein expression of p-p38 MAPK and p-JNK was significantly lower than that of km group. Conclusion P38 MAPK and JNK mediated the ototoxic injury induced by km in BALB/c mice and the high expression of p-p38 MAPK and p-JNK was significantly inhibited by p38 MAPK and JNK. Thus, the ototoxicity of km can be effectively protected, which may be one of the protective mechanisms of LA.
【學(xué)位授予單位】：遼寧醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R764.43;R965

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 馬慧敏;周安國(guó);王之盛;馬志勇;;硫辛酸的抗氧化功能及其在動(dòng)物生產(chǎn)上的應(yīng)用現(xiàn)狀[J];飼料工業(yè);2006年22期

2 于利;湯浩;王愛梅;;卡那霉素所致小鼠耳蝸細(xì)胞凋亡途徑的研究[J];廣東醫(yī)學(xué);2011年01期

3 張t盼

本文編號(hào)：1445745