【摘要】：目的探討大鼠早期壓瘡缺血再灌注損傷的凋亡機(jī)制及曲美他嗪預(yù)處理的效果,為臨床早期壓瘡的防治提供基礎(chǔ)理論依據(jù)和新思路。方法1將16只Wistar大鼠隨機(jī)分為2組,對照組(Sham組)和缺血/再灌注組(I/R組)。I/R組大鼠應(yīng)用特制壓力裝置制備大鼠早期壓瘡缺血再灌注損傷模型。實(shí)驗(yàn)終點(diǎn)取受壓局部肌肉組織,采用HE染色法觀察大鼠受壓肌肉組織的病理學(xué)變化;TUNEL法檢測大鼠受壓組織中肌細(xì)胞凋亡情況;免疫組織化學(xué)法檢測受壓肌肉組織中凋亡相關(guān)蛋白Bax與Bcl-2的表達(dá)情況;Western blot法檢測凋亡相關(guān)蛋白Bax、Bcl-2和CHOP在受壓組織中的表達(dá)水平。2將40只Wistar大鼠隨機(jī)分為5組,對照組(Sham組)、缺血/再灌注組(I/R組)、曲美他嗪低劑量組(TMZ低組)、曲美他嗪中劑量組(TMZ中組)、曲美他嗪高劑量組(TMZ高組)。應(yīng)用特制壓力裝置制備大鼠壓瘡缺血再灌注損傷模型。I/R組大鼠只造模不給藥,曲美他嗪預(yù)處理組大鼠造模前7天分別給予曲美他嗪低(5mg/kg)、中(10mg/kg)、高(15mg/kg)劑量灌胃預(yù)處理。實(shí)驗(yàn)終點(diǎn)取受壓局部肌肉組織,采用HE染色法觀察各組大鼠受壓肌肉組織的病理學(xué)變化;TUNEL法檢測各組大鼠受壓組織中肌細(xì)胞凋亡情況;免疫組織化學(xué)法檢測各組受壓肌肉組織中凋亡相關(guān)蛋白Bax與Bcl-2的表達(dá)情況;Western blot法檢測凋亡相關(guān)蛋白Bax、Bcl-2和CHOP在受壓組織中的表達(dá)水平。結(jié)果1光鏡觀察受壓組織發(fā)現(xiàn),Sham組肌纖維排列緊密,形態(tài)正常,無炎性細(xì)胞浸潤及病理改變,與Sham組相比I/R組肌纖維病理損傷嚴(yán)重;TUNEL法檢測受壓組織凋亡結(jié)果顯示,Sham組中幾乎很少見到凋亡細(xì)胞,I/R組凋亡細(xì)胞的表達(dá)顯著增多(P0.05);免疫組化檢測Bax和Bcl-2蛋白結(jié)果與Western blot結(jié)果趨勢一致,即Bax蛋白在Sham組中幾乎不表達(dá)或有極少表達(dá),與Sham組相比I/R組Bax蛋白表達(dá)顯著升高(P0.05),Bcl-2蛋白在Sham組中表達(dá)顯著,在I/R組中的表達(dá)與Sham組相比顯著降低(P0.05);Western blot法檢測受壓組織中CHOP的表達(dá)結(jié)果顯示,在Sham組中幾乎不表達(dá)或僅有極少表達(dá),與Sham組相比I/R組CHOP蛋白表達(dá)顯著升高(P0.05)。2光鏡觀察受壓組織發(fā)現(xiàn),Sham組肌纖維排列緊密,形態(tài)正常,與Sham組相比I/R組肌纖維病理損傷嚴(yán)重,在曲美他嗪預(yù)處理組中隨著給藥劑量的增加病理損傷逐漸減輕。TUNEL法檢測受壓組織凋亡結(jié)果顯示,Sham組中幾乎很少見到凋亡細(xì)胞,I/R組凋亡細(xì)胞的表達(dá)顯著增多(P0.05),在曲美他嗪預(yù)處理組中隨著給藥劑量的增加凋亡細(xì)胞表達(dá)逐漸減少。免疫組化檢測Bax和Bcl-2蛋白結(jié)果與Western blot結(jié)果趨勢一致,即Bax蛋白在Sham組中幾乎不表達(dá)或有極少表達(dá),與Sham組相比I/R組Bax蛋白表達(dá)顯著升高(P0.05),在曲美他嗪預(yù)處理組中隨著給藥劑量的增加表達(dá)逐漸減少,Bcl-2蛋白在Sham組中表達(dá)顯著,在I/R組中的表達(dá)與Sham組相比顯著降低(P0.05),在曲美他嗪預(yù)處理組中隨著給藥劑量的增加表達(dá)逐漸增多。Western blot法檢測受壓組織中CHOP的表達(dá)結(jié)果顯示,在Sham組中幾乎不表達(dá)或僅有極少表達(dá),與Sham組相比I/R組CHOP蛋白表達(dá)顯著升高(P0.05),在曲美他嗪預(yù)處理組中隨著給藥劑量的增加表達(dá)逐漸減少。結(jié)論1細(xì)胞凋亡參與了大鼠早期壓瘡的形成,受壓組織中Bcl-2蛋白表達(dá)減少,Bax和CHOP蛋白表達(dá)增多。2曲美他嗪預(yù)處理對大鼠早期壓瘡缺血再灌注損傷具有保護(hù)作用,可以減輕受壓肌肉組織的病理損傷程度,上調(diào)Bcl-2表達(dá)水平,下調(diào)Bax、CHOP表達(dá)水平抑制細(xì)胞凋亡,保護(hù)受壓肌肉組織。曲美他嗪高劑量(15mg/kg)對大鼠早期壓瘡缺血再灌注損傷的保護(hù)效果較低劑量好。
[Abstract]:Objective To investigate the mechanism of apoptosis in early stage pressure sore of rats and the pre-treatment effect of Qu Mei, and to provide theoretical basis and new thought for the prevention and cure of early stage pressure sore. Methods 16 Wistar rats were randomly divided into 2 groups, control group (sham group) and ischemia/ reperfusion group (I/ R group). A model of ischemia-reperfusion injury in rats with early pressure sore was prepared by using special pressure device in group I/ R rats. The results showed that the expression of Bax and Bcl-2 in compressed muscle tissue was detected by HE staining, and the expression of Bax and Bcl-2 was detected by immunohistochemical method. The expression levels of Bax, Bcl-2 and CHOP in compressed tissues were detected by Western blot. Forty Wistar rats were randomly divided into 5 groups, the control group (sham group), ischemia/ reperfusion group (I/ R group), the low-dose group (TMZ low group), and the dose group (group TMZ in TMZ). He was a high dose group (TMZ high). A model of ischemia-reperfusion injury in rats with pressure sore was prepared by using special pressure device. Rats in the I/ R group were not dosed, and the pre-treatment group was given a dose of low (5mg/ kg), medium (10mg/ kg) and high (15mg/ kg), respectively. At the end of the experiment, the local muscle tissue was compressed, and the pathological changes of muscle tissue were observed by HE staining, and the apoptosis was detected by TUNEL method. The expression of Bax, Bcl-2 and Bax, Bcl-2 and CHOP were detected by Western blot. Results According to light microscope, it was found that the muscle fibers of Shaam group were closely packed, the morphology was normal, there was no inflammatory cell infiltration and pathological change. Compared with the sham group, the pathological changes of myofibers in the I/ R group were serious. The expression of Bax and Bcl-2 protein in I/ R group was significantly increased (P0.05). The expression of Bcl-2 protein in group I/ R was significantly lower than that of sham group (P0.05). The expression of CHOP in compressed tissues was detected by Western blot. The expression of CHOP protein in group I/ R group was significantly higher than that of sham group (P0.05). The expression of apoptotic cells in the group I/ R was significantly increased (P0.05). The expression of Bax and Bcl-2 protein was similar to that of Western blot, that is, the expression of Bax protein was hardly expressed or expressed in the sham group, and the expression of Bax protein in the group I/ R was significantly higher than that of the sham group (P0.05). The expression of Bcl-2 protein in the sham group was significant, and the expression of Bcl-2 in the I/ R group was significantly lower than that of the sham group (P0.05). The expression of CHOP in compressed tissues was detected by Western blot. The expression of CHOP protein in group I/ R was significantly higher than that of sham group (P0.05). Conclusion 1 Apoptosis is involved in the formation of early pressure sore in rats, the expression of Bcl-2 protein in compressed tissues decreases, the expression of Bax and CHOP protein is increased. The expression level of Bcl-2 was up-regulated, the expression level of Bax and CHOP was down-regulated to inhibit cell apoptosis and to protect the compressed muscle tissue. The effect of high dose (15mg/ kg) on ischemia-reperfusion injury of early stage pressure sore in rats was lower than that of low dose.
【學(xué)位授予單位】：華北理工大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R47

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