本文關(guān)鍵詞： 電針刺 內(nèi)毒素 急性肺損傷 內(nèi)質(zhì)網(wǎng)應(yīng)激 氧化應(yīng)激　出處：《天津醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：急性肺損傷(ALI)是臨床上常見的疾病,多數(shù)患者由于感染、外傷、休克等引起,發(fā)病后臨床表現(xiàn)為中性粒細胞浸潤,嚴重者可發(fā)展為急性呼吸窘迫綜合征(acute respiratory distress syndrome,ARDS),甚至造成多器官功能障礙綜合征(multiple organ dysfunctions syndrome,MODS)等嚴重并發(fā)癥。在臨床上有較高的發(fā)病率和死亡率,目前對此尚無理想的治療手段[1-3]。內(nèi)質(zhì)網(wǎng)是真核生物重要的細胞器,是細胞蛋白合成、加工、折疊、運送及鈣離子儲存的主要場所[4]。內(nèi)質(zhì)網(wǎng)具有維持細胞內(nèi)環(huán)境穩(wěn)態(tài)平衡的重要作用[5]。而內(nèi)質(zhì)網(wǎng)應(yīng)激(endoplasmic reticulum stress,ERS)作為細胞對外界不良刺激的自身保護反應(yīng),可引起凋亡相關(guān)基因如葡萄糖調(diào)節(jié)蛋白78(GRP78)、CCAAT/增強子結(jié)合蛋白同源蛋白(CHOP)、caspase-12等表達上調(diào),從而啟動細胞凋亡,內(nèi)質(zhì)網(wǎng)應(yīng)激(ERS)可能是肺部相關(guān)疾病關(guān)鍵的病理生理機制之一[6-9]。我們前期研究已經(jīng)證實了電針刺激對內(nèi)毒素所致的急性肺損傷(ALI)具有防治作用,但其對內(nèi)質(zhì)網(wǎng)應(yīng)激(ERS)的具體作用尚不明確[10-13]。目的本課題擬從內(nèi)質(zhì)網(wǎng)應(yīng)激(ERS)角度探討內(nèi)毒素急性肺損傷(ALI)時電針刺激在微環(huán)境層面的內(nèi)源性分子保護作用機制,為電針刺激在臟器保護的機制研究開辟新的思路,也為電針刺激防治內(nèi)毒素性急性肺損傷(ALI)提供新的理論依據(jù)。方法40只健康清潔級雄性SD大鼠,8周齡,隨機數(shù)字表法分為對照組(C組)、內(nèi)毒素急性肺損傷模型組(LA組)、電針刺激穴位+內(nèi)毒素急性肺損傷模型組(ELA組)、非穴位電針刺激+內(nèi)毒素急性肺損傷模型組(NLA組),每組10只。LA組、NLA組和ELA組大鼠將內(nèi)毒素脂多糖(lipopolysaccharide,LPS)5mg/kg溶于0.5m L生理鹽水經(jīng)尾靜脈注射,建立大鼠ALI模型;C組給予等容量生理鹽水。ELA組于內(nèi)毒素急性肺損傷(ALI)模型制備前1~4d及模型制備過程中電針刺激雙側(cè)足三里和內(nèi)關(guān)穴(疏密波,頻率2/15Hz,刺激電流1~2m A,波寬0.2~0.6ms,刺激強度以大鼠肢體出現(xiàn)輕微顫動為宜),1次/d,30min/次,刺激時間為上午9:30-10:30;NLA組以相同的參數(shù)電針刺激足三里和內(nèi)關(guān)穴旁開0.5cm非經(jīng)非穴位處。靜脈注射脂多糖(LPS)或生理鹽水后6h時,處死大鼠留取肺組織處理后保存。進行肺組織病理學(xué)觀察及損傷評分;肺組織濕重/干重比值(W/D比值)測定;TUNEL法檢測肺泡上皮細胞凋亡指數(shù)(AI);Western blot法檢測肺組織葡萄糖調(diào)節(jié)蛋白78(GRP78)、CHOP及caspase-12蛋白表達水平。結(jié)果與C組比較,LA組、NLA組及ELA組肺損傷評分、W/D比值及肺泡上皮細胞凋亡指數(shù)(AI)升高,肺組織GRP78、CHOP和caspase-12蛋白表達水平上調(diào)(P0.05);與LA組比較,ELA組肺損傷評分、濕重/干重比值(W/D比值)和肺泡上皮細胞凋亡指數(shù)(AI)降低,肺組織GRP78、CHOP及caspase-12蛋白水平下降(P0.05),NLA組以上指標比較差異無統(tǒng)計學(xué)意義(P0.05)。結(jié)論電針刺激雙側(cè)足三里和內(nèi)關(guān)穴可減輕大鼠內(nèi)毒素致急性肺損傷(ALI),其機制與抑制大鼠肺組織內(nèi)質(zhì)網(wǎng)應(yīng)激(ERS)有關(guān)。
[Abstract]:Acute lung injury (Ali) is a common clinical disease. Most of the patients suffer from infection, trauma, shock and so on. In severe cases, acute respiratory distress syndrome may develop into ARDS, and even cause multiple organ dysfunctions syndrome, such as mods. The endoplasmic reticulum (ER) is an important organelle of eukaryotes, which is the synthesis, processing and folding of cellular proteins. Major sites for transport and storage of calcium ions [4] .Endoplasmic reticulum (ER) plays an important role in maintaining homeostasis of intracellular environment [5]. Endoplasmic reticulum stress (ER) is the self-protective response of cells to adverse external stimuli. The expression of apoptosis-related genes such as Glucose-regulated protein 78, GRP78, CCAATA / enhancer binding protein homologue protein, CHOPP7, caspase-12, was up-regulated, thus inducing apoptosis. Endoplasmic reticulum stress (ERS) may be one of the key pathophysiological mechanisms of lung related diseases [6-9]. Our previous studies have confirmed that electroacupuncture has a preventive and therapeutic effect on endotoxin-induced acute lung injury (ALI). But the specific effect of ERS on endoplasmic reticulum stress (ERS) is unclear [10-13]. Objective to explore the mechanism of endogenous molecular protection of electroacupuncture stimulation in microenvironment from the perspective of endoplasmic reticulum stress (ERS). This study provided a new theoretical basis for the study of the mechanism of electroacupuncture stimulation in organ protection and for the prevention and treatment of endotoxin induced acute lung injury (ALII). Methods 40 healthy and clean male Sprague-Dawley rats were 8 weeks old. The method was randomly divided into three groups: control group (C group), endotoxin acute lung injury model group (LA group), electroacupuncture stimulation point endotoxin acute lung injury model group (ELA group) and non-acupoint electroacupuncture stimulation endotoxin acute lung injury model group (NLA group). 10 rats in each group were injected intravenously with lipopolysaccharide of lipopolysaccharide of lipopolysaccharide (LPS5 mg / kg) dissolved in 0.5 mL of normal saline in each group (n = 10) and ELA group (n = 10). Rats in ALI model group C were given isovolumic physiological saline. ELA group was treated with endotoxin-induced acute lung injury (Ali) 1 day before the establishment of the model and electroacupuncture stimulated bilateral Zusanli and Neiguan points (dense wave) during the preparation of the model. At the frequency of 2 / 15 Hz, the stimulation current is 12mA, the wave width is 0.2ms, and the stimulation intensity is as follows: slight tremor in the limbs of the rat is suitable for 30 mins per rout. The stimulation time was 9: 30-10: 30: 30: 30: NLA group with the same parameters to stimulate Zusanli and Neiguan acupoints with 0.5 cm open at non-acupoints. 6 hours after intravenous injection of lipopolysaccharide (LPS) or normal saline, The rats were sacrificed and preserved after the treatment of lung tissue. The histopathological observation and injury score of the lung were performed. Lung tissue wet weight / dry weight ratio (WR / D) was used to detect apoptosis index of alveolar epithelial cells by Tunel method and Western blot method to detect the expression of glucose-regulated protein 78GRP 78 and caspase-12 protein in lung tissue. Results compared with group C, the expression of glucose regulated protein 78 GRP 78 and caspase-12 protein were detected in LA group and ELA group. The ratio of W / D and the apoptosis index of alveolar epithelial cells were increased. Compared with LA group, lung injury score, wet / dry weight ratio (WR / D) and apoptosis index of alveolar epithelial cells (AI) were decreased. There was no significant difference in the above indexes between the two groups (P 0.05). Conclusion the stimulation of bilateral Zusanli and Neiguan points by electroacupuncture can reduce the acute lung injury induced by endotoxin in rats. Endoplasmic reticulum stress (ERS) is involved.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R563.8

【參考文獻】

相關(guān)期刊論文 前8條

1 李英娜;劉志建;佟德惠;;七氟醚預(yù)處理對肢體缺血再灌注誘發(fā)大鼠肺損傷的影響及其與內(nèi)質(zhì)網(wǎng)應(yīng)激的關(guān)系[J];中華麻醉學(xué)雜志;2016年05期

2 王穎;王丹;余劍波;宮麗榮;張圓;董樹安;穆蕊;史佳;劉大全;;線粒體融合-分裂在大鼠內(nèi)毒素性急性肺損傷中的作用[J];中華麻醉學(xué)雜志;2015年05期

3 徐晶晶;王志萍;;內(nèi)質(zhì)網(wǎng)應(yīng)激在大鼠肺缺血再灌注損傷中的作用[J];中華麻醉學(xué)雜志;2015年04期

4 宮麗榮;余劍波;徐妍;曹新順;史佳;張桂誠;;活化蛋白-1在電針上調(diào)內(nèi)毒素休克兔肺組織HO-1表達中的作用[J];中華麻醉學(xué)雜志;2014年03期

5 姚樹桐;秦樹存;;內(nèi)質(zhì)網(wǎng)應(yīng)激在動脈粥樣硬化發(fā)生、發(fā)展和防治中的作用[J];中國病理生理雜志;2014年02期

6 張圓;余劍波;宮麗榮;王曼;曹新順;閆雨苗;董樹安;唐林;;NF-κB在電針上調(diào)內(nèi)毒素休克誘發(fā)急性肺損傷兔血紅素氧合酶-1表達中的作用[J];中華麻醉學(xué)雜志;2013年08期

7 伍熙;柳景華;;內(nèi)質(zhì)網(wǎng)應(yīng)激與動脈粥樣硬化治療[J];中華老年心腦血管病雜志;2013年07期

8 蔣莉婭;黃繼人;趙弘卿;張衛(wèi)東;戴建良;;“肺與大腸相表里”在針刺治療重癥急性胰腺炎大鼠早期急性肺損傷中應(yīng)用的實驗研究[J];中國中醫(yī)基礎(chǔ)醫(yī)學(xué)雜志;2011年11期

，

本文編號：1555212