丁酸鈉對(duì)慶大霉素耳毒性保護(hù)機(jī)制的體內(nèi)研究
發(fā)布時(shí)間：2019-06-04 09:15
【摘要】：實(shí)驗(yàn)一：凝膠海綿圓窗龕局部給藥對(duì)正常豚鼠聽力的影響目的：研究經(jīng)手術(shù)，圓窗龕局部放置凝膠海綿的給藥方式對(duì)正常豚鼠的聽性腦干反應(yīng)（AuditorybrainstemresponseABR）閾值的影響。 方法：5只健康紅目白化豚鼠，單耳手術(shù)。顯微鉆打開聽泡,暴露圓窗,取0.3mm~3凝膠海綿吸取10ul外淋巴液放置于圓窗龕。術(shù)后立即行ABR檢測(cè)。了解通過手術(shù)，圓窗龕局部放置凝膠海綿的給藥方式對(duì)正常豚鼠ABR閾值的影響。 結(jié)果：配對(duì)T檢驗(yàn)：顯示手術(shù)前后ABR的反應(yīng)閾值有統(tǒng)計(jì)學(xué)差異（P0.01）。 結(jié)論：本實(shí)驗(yàn)證實(shí)了，圓窗龕放置凝膠海綿的給藥方式，因?yàn)楦淖兞苏５膫鲗?dǎo)方式，故而影響了正常豚鼠的ABR反應(yīng)閾值。所以，在接下來的實(shí)驗(yàn)中，ABR閾值的檢測(cè)結(jié)果的比較要考慮到給藥方式本身的影響。 實(shí)驗(yàn)二：丁酸鈉圓窗給藥毒性研究 目的：了解組蛋白去乙�；敢种苿℉DACi）丁酸鈉（NaB）圓窗龕給藥對(duì)毛細(xì)胞有無(wú)毒性作用。 方法:5只紅目白化豚鼠單耳手術(shù)，圓窗龕放置0.3mm~3凝膠海綿，吸附10ul丁酸鈉NaB(100mg/ml)，局部放置14天。而后行ABR檢測(cè)、基底膜鋪片、掃描電鏡。觀察丁酸鈉對(duì)毛細(xì)胞有無(wú)毒性作用。 結(jié)果：?jiǎn)我蛩胤讲罘治鲲@示：給藥后ABR閾值與給藥后-給藥前的ABR閾移，與單純手術(shù)組相比無(wú)統(tǒng)計(jì)學(xué)意義（P0.05）。全基底膜鋪片顯示：毛細(xì)胞無(wú)缺失。掃描電鏡顯示：纖毛形態(tài)正常。 結(jié)論：丁酸鈉對(duì)正常豚鼠的聽力，以及毛細(xì)胞無(wú)損傷作用。本實(shí)驗(yàn)證實(shí)圓窗膜龕放置丁酸鈉無(wú)毒性損傷作用。故可采用圓窗膜給藥的方式進(jìn)一步驗(yàn)證其是否對(duì)慶大霉素耳毒性具有保護(hù)作用。 實(shí)驗(yàn)三：丁酸鈉對(duì)慶大霉素耳毒性的保護(hù)作用 目的：研究丁酸鈉對(duì)慶大霉素耳毒性的保護(hù)作用。 方法：10只雄性紅目白化豚鼠，實(shí)驗(yàn)期間，可以自由攝取食物與水。經(jīng)ABR檢測(cè)聽力正常后，雙耳手術(shù)。通過手術(shù)方法向置放在豚鼠右耳圓窗龕內(nèi)約0.3mm~3的明膠海綿注入10ul丁酸鈉（100mg/ml）溶液；向置放在豚鼠左耳圓窗龕內(nèi)約0.3mm~3的明膠海綿注入10ul外淋巴液作為空白對(duì)照。3天后肌注慶大霉素（200mg/kg.day連續(xù)5天）.休息3天行ABR檢測(cè)、基底膜鋪片染色、掃描電鏡觀察、毛細(xì)胞缺失率計(jì)數(shù)及免疫熒光染色觀察丁酸鈉是否對(duì)慶大霉素耳毒性具有保護(hù)作用。 結(jié)果：配對(duì)T檢驗(yàn)顯示：丁酸鈉組ABR閾值與外淋巴液組相比較有統(tǒng)計(jì)學(xué)差異（P0.01)�；啄や伷�，鬼筆環(huán)肽染色示：第二轉(zhuǎn)丁酸鈉組毛細(xì)胞缺失較外淋巴液組少。掃描電鏡觀察：對(duì)照組毛細(xì)胞缺失嚴(yán)重，特別是第二、三層外毛細(xì)胞。殘存的毛細(xì)胞纖毛腫脹、倒伏，排列紊亂，失去毛細(xì)胞之間的正常連接。同時(shí)，內(nèi)毛細(xì)胞亦有缺失，伴排列紊亂，纖毛倒伏、腫脹。給予保護(hù)劑丁酸鈉后外毛細(xì)胞缺失明顯減少，纖毛腫脹也較對(duì)照組減輕。毛細(xì)胞缺失率計(jì)數(shù)顯示：對(duì)照組，可見外毛細(xì)胞距頂轉(zhuǎn)約10-40%即有減少，距頂轉(zhuǎn)約55%開始明顯減少，距頂轉(zhuǎn)約70%基本無(wú)外毛細(xì)胞存在。內(nèi)毛細(xì)胞距頂轉(zhuǎn)約60%開始減少，70%處明顯減少，80%處基本無(wú)內(nèi)毛細(xì)胞存在。給予保護(hù)劑丁酸鈉組可見外毛細(xì)胞距頂轉(zhuǎn)約55-65%即有減少，距頂轉(zhuǎn)約70%開始明顯減少，距頂轉(zhuǎn)約80%基本無(wú)外毛細(xì)胞存在。內(nèi)毛細(xì)胞距頂轉(zhuǎn)約70%開始減少，75%處明顯減少，80%處基本無(wú)內(nèi)毛細(xì)胞存在。免疫熒光檢測(cè)示:1.正常豚鼠圓窗龕給予丁酸鈉（NaB）作用后，HDAC1的表達(dá)減低。 2.隨著慶大霉素作用時(shí)間的延長(zhǎng)，HDAC1的熒光亮度增加，表達(dá)量增高。3.形態(tài)異常的核，HDAC1的表達(dá)是增加的。4.慶大霉素作用后5天，出現(xiàn)核固縮、溶解、消失。HDAC1由于是核表達(dá)，故表現(xiàn)出同樣的變化。5.給予丁酸鈉組慶大霉素作用后5天。雖然也有核的溶解、消失，但多數(shù)核的形態(tài)正常。且HDAC1的熒光亮度較對(duì)照組低，表達(dá)量較對(duì)照組減少。結(jié)論：組蛋白去乙�；敢种苿�，，丁酸鈉（NaB）對(duì)慶大霉素耳毒性具有保護(hù)作用。并且此保護(hù)作用是通過抑制組蛋白的去乙�；蕉鴮�(shí)現(xiàn)的。
[Abstract]:Objective: To study the effect of local administration of gel sponge round window niche on the hearing of the normal guinea pig: study the effect of the administration of local gel sponge on the threshold of the auditory brainstem response (ABR) of the normal guinea pig. Method:5 healthy red eyes albino guinea pigs, single ear Intraoperative. The micro-drill opened the sound bulb and exposed the round window, and the 0.3 mm ~ 3 gel sponge was used to draw the 10 ul of the external lymph fluid to the round window. niches. The ABR test was performed immediately after the operation Measurement. See the shadow of the normal guinea pig's ABR threshold by means of the procedure and the local placement of the gel sponge in the round window niche. Response: paired T-test: There was a statistical difference in the response threshold of the ABR before and after the operation (P0. 01). Conclusion: The experimental results confirm that the round window niche is used to place the gel sponge, because the normal conduction mode is changed, thus the AB of the normal guinea pig is affected. R reaction threshold. Therefore, in the next experiment, the comparison of the detection results of the ABR threshold should take into account the mode of administration The effect of sodium butyrate on its own. The purpose of the study on the toxicity of round-window administration: to understand the administration of histone to the niches (NaB) round-window niche of histone-deethanolase inhibitor (HDACi). The hair cells were non-toxic. Methods:5 eyes with albino guinea pigs were operated in a single ear, 0.3 mm ~ 3 gel sponge was placed in the round window niche, and NaB (100 mg/ kg) was adsorbed. ml), localized for 14 days, followed by an ABR test And scanning electron microscope to observe the butyric acid. The effect of sodium on hair cells was non-toxic. Results: The single-factor analysis of variance showed that the ABR threshold after administration and the ABR threshold after administration-predose were compared to the simple surgical group. No statistical significance (P0.05). The hair cells showed no missing hair cells. Scanning electron microscope (SEM) showed that the ciliated form was normal. The hearing of the mouse, as well as the damage of the hair cell, confirmed the circle The window film niche is used for the non-toxic damage of sodium butyrate, so that the round window membrane can be used for further verifying that it is No protective effect on the ototoxicity of gentamicin. The protective effect of sodium butyrate on the ototoxicity of gentamicin : To study the protective effect of sodium butyrate on the ototoxicity of gentamicin. in that albino guinea pig, free uptake during the course of the experiment The food and water were tested by the ABR. After the hearing was normal, the two-ear operation was performed. The solution of 10 ul of sodium butyrate (100 mg/ ml) was injected into the gelatin sponge placed in the round window niche of the right ear of the guinea pig through the surgical method, and 10 ul of the external lymph fluid was injected into the gelatin sponge placed in the round window niche of the left ear of the guinea pig to be used as the blank control. After 3 days, the gentamicin (200 mg/ < Chunk> kg. day 5 consecutive days). ABR detection, basement membrane spreading, scanning electron microscopy, hair cell loss rate count and immunofluorescence staining for 3 days of rest. The protective effect of sodium butyrate on the ototoxicity of gentamicin was observed. Results: The test of paired T-test showed that the ABR threshold of sodium butyrate group There was a significant difference between the value and the external lymph group (P0.01). The results showed that the hair cells of the second sodium butyrate group were less than that of the external lymph fluid group. The hair cells of the control group were missing, especially the second and the three outer hair cells. The remaining hair cells were cilia. Swelling, lodging, disorder, and loss of normal connections between hair cells. And the inner hair cells are also deleted, and the hair cells are arranged in a disorder, the cilia lodging and the swelling are arranged, and sodium butyrate is administered. The loss of hair cells in the outer hair cells was significantly reduced, and the cilia swelling was also reduced in the control group. The counting of the loss rate of the hair cells showed that the control group, the visible outer hair cell was about 10-40% from the top to the top, and the distance from the top to the top was about 55%. There was a significant reduction in the beginning, about 70% from the top and about 70% in the absence of an outer hair cell. The inner hair cell started to decrease from the top to about 60%. There is a significant reduction in 70%, and there is no internal hair cell at 80%. It can be seen that the hair cell is reduced from the top to about 55-65% from the top to about 70% from the top. There was a significant reduction in the onset of the primary hair cell from the top to about 80%. The inner hair cell was reduced from the top to about 70%. at 75%, There are basically no internal hair cells at 80%. Immunofluorescence detection:1. The normal guinea pig round window niche The expression of HDAC1 decreased after the effect of sodium butyrate (NaB). The increase of the fluorescence intensity of HDAC1 and the increase of the expression of HDAC1 were increased. High.3. The expression of HDAC1 is increased in the nucleus with abnormal morphology. .4. After 5 days after the effect of gentamycin, the presence of the nuclear solid, the dissolution and the elimination Loss. HDAC1 is a core expression, so the table the same changes were shown.5. The effect of gentamicin in the sodium butyrate group After 5 days, although there was also the dissolution of the nucleus, it disappeared, but the majority of the nuclei were in normal form. The fluorescence intensity of HDAC1 was lower than that of the control group. The expression of HDAC1 was lower than that in the control group. The preparation and sodium butyrate (NaB) have a protective effect on the ototoxicity of gentamicin.
【學(xué)位授予單位】：第四軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R764
本文編號(hào)：2492624
[Abstract]:Objective: To study the effect of local administration of gel sponge round window niche on the hearing of the normal guinea pig: study the effect of the administration of local gel sponge on the threshold of the auditory brainstem response (ABR) of the normal guinea pig. Method:5 healthy red eyes albino guinea pigs, single ear Intraoperative. The micro-drill opened the sound bulb and exposed the round window, and the 0.3 mm ~ 3 gel sponge was used to draw the 10 ul of the external lymph fluid to the round window. niches. The ABR test was performed immediately after the operation Measurement. See the shadow of the normal guinea pig's ABR threshold by means of the procedure and the local placement of the gel sponge in the round window niche. Response: paired T-test: There was a statistical difference in the response threshold of the ABR before and after the operation (P0. 01). Conclusion: The experimental results confirm that the round window niche is used to place the gel sponge, because the normal conduction mode is changed, thus the AB of the normal guinea pig is affected. R reaction threshold. Therefore, in the next experiment, the comparison of the detection results of the ABR threshold should take into account the mode of administration The effect of sodium butyrate on its own. The purpose of the study on the toxicity of round-window administration: to understand the administration of histone to the niches (NaB) round-window niche of histone-deethanolase inhibitor (HDACi). The hair cells were non-toxic. Methods:5 eyes with albino guinea pigs were operated in a single ear, 0.3 mm ~ 3 gel sponge was placed in the round window niche, and NaB (100 mg/ kg) was adsorbed. ml), localized for 14 days, followed by an ABR test And scanning electron microscope to observe the butyric acid. The effect of sodium on hair cells was non-toxic. Results: The single-factor analysis of variance showed that the ABR threshold after administration and the ABR threshold after administration-predose were compared to the simple surgical group. No statistical significance (P0.05). The hair cells showed no missing hair cells. Scanning electron microscope (SEM) showed that the ciliated form was normal. The hearing of the mouse, as well as the damage of the hair cell, confirmed the circle The window film niche is used for the non-toxic damage of sodium butyrate, so that the round window membrane can be used for further verifying that it is No protective effect on the ototoxicity of gentamicin. The protective effect of sodium butyrate on the ototoxicity of gentamicin : To study the protective effect of sodium butyrate on the ototoxicity of gentamicin. in that albino guinea pig, free uptake during the course of the experiment The food and water were tested by the ABR. After the hearing was normal, the two-ear operation was performed. The solution of 10 ul of sodium butyrate (100 mg/ ml) was injected into the gelatin sponge placed in the round window niche of the right ear of the guinea pig through the surgical method, and 10 ul of the external lymph fluid was injected into the gelatin sponge placed in the round window niche of the left ear of the guinea pig to be used as the blank control. After 3 days, the gentamicin (200 mg/ < Chunk> kg. day 5 consecutive days). ABR detection, basement membrane spreading, scanning electron microscopy, hair cell loss rate count and immunofluorescence staining for 3 days of rest. The protective effect of sodium butyrate on the ototoxicity of gentamicin was observed. Results: The test of paired T-test showed that the ABR threshold of sodium butyrate group There was a significant difference between the value and the external lymph group (P0.01). The results showed that the hair cells of the second sodium butyrate group were less than that of the external lymph fluid group. The hair cells of the control group were missing, especially the second and the three outer hair cells. The remaining hair cells were cilia. Swelling, lodging, disorder, and loss of normal connections between hair cells. And the inner hair cells are also deleted, and the hair cells are arranged in a disorder, the cilia lodging and the swelling are arranged, and sodium butyrate is administered. The loss of hair cells in the outer hair cells was significantly reduced, and the cilia swelling was also reduced in the control group. The counting of the loss rate of the hair cells showed that the control group, the visible outer hair cell was about 10-40% from the top to the top, and the distance from the top to the top was about 55%. There was a significant reduction in the beginning, about 70% from the top and about 70% in the absence of an outer hair cell. The inner hair cell started to decrease from the top to about 60%. There is a significant reduction in 70%, and there is no internal hair cell at 80%. It can be seen that the hair cell is reduced from the top to about 55-65% from the top to about 70% from the top. There was a significant reduction in the onset of the primary hair cell from the top to about 80%. The inner hair cell was reduced from the top to about 70%. at 75%, There are basically no internal hair cells at 80%. Immunofluorescence detection:1. The normal guinea pig round window niche The expression of HDAC1 decreased after the effect of sodium butyrate (NaB). The increase of the fluorescence intensity of HDAC1 and the increase of the expression of HDAC1 were increased. High.3. The expression of HDAC1 is increased in the nucleus with abnormal morphology. .4. After 5 days after the effect of gentamycin, the presence of the nuclear solid, the dissolution and the elimination Loss. HDAC1 is a core expression, so the table the same changes were shown.5. The effect of gentamicin in the sodium butyrate group After 5 days, although there was also the dissolution of the nucleus, it disappeared, but the majority of the nuclei were in normal form. The fluorescence intensity of HDAC1 was lower than that of the control group. The expression of HDAC1 was lower than that in the control group. The preparation and sodium butyrate (NaB) have a protective effect on the ototoxicity of gentamicin.
【學(xué)位授予單位】：第四軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R764
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