本文選題：電針 + 腦缺血再灌注��； 參考：《南京中醫(yī)藥大學(xué)》2017年碩士論文

【摘要】：目的:通過觀察電針對腦缺血再灌注大鼠模型炎癥反應(yīng)相關(guān)因子TNF-α、IL-4的調(diào)節(jié)作用,探討電針改善腦缺血后大鼠炎癥反應(yīng)的作用機(jī)制,以期進(jìn)一步深化電針治療缺血性腦病作用機(jī)理的認(rèn)識,為臨床治療提供科學(xué)依據(jù)。方法:雄性清潔級SD大鼠,采用數(shù)字表法隨機(jī)抽取24只作為假手術(shù)組,其余剩下的動物采用改良Longa線栓法制備大鼠右側(cè)大腦中動脈腦缺血模型,根據(jù)Zea-longa5級4分法的評分標(biāo)準(zhǔn),初步判斷造模成功與否,將成功的模型隨機(jī)分為模型組和電針組,每組有效例數(shù)為24只。假手術(shù)組只分離血管不穿插線栓;模型組用線栓阻塞中動脈2h后拔出線栓實現(xiàn)再灌注,之后不做任何治療措施;電針組成功造模后選取"百會"和"大椎"穴進(jìn)行針刺,連接韓式HANS-200型電針儀治療30min,波形為疏密波,刺激頻率為2/15Hz。各組于缺血再灌注24h后,參照J(rèn)ulio氏18分制評分法行神經(jīng)行為學(xué)評分;采用TTC染色法觀察腦缺血組織形態(tài)學(xué)改變和腦梗死面積百分比;采用蘇木素-伊紅(HE)染色法觀察缺損側(cè)腦組織細(xì)胞病理變化;采用免疫組化染色法檢測缺損側(cè)皮層組織IL-4、TNF-α的蛋白表達(dá);采用 qRT-PCR 檢測 IL-4mRNA、TNF-αmRNA 表達(dá)的變化。結(jié)果:(1)神經(jīng)功能行為學(xué)評分:腦缺血再灌注損傷大鼠的神經(jīng)功能行為學(xué)評分明顯降低,與假手術(shù)組相比差異有統(tǒng)計學(xué)意義(P0.01);與模型組比較,電針干預(yù)治療能有效提高神經(jīng)行為學(xué)評分,差異有統(tǒng)計學(xué)意義(P0.05)。(2)腦組織形態(tài)學(xué)變化及梗死面積百分比:經(jīng)TTC染色,假手術(shù)組腦組織均勻染色呈現(xiàn)暗紅色,模型組大腦右側(cè)缺血腦組織呈現(xiàn)白色,電針組染色缺失區(qū)較模型組減少。模型組和電針組腦梗死面積百分比均高于假手術(shù)組(P0.01和P0.05),與模型組比較,電針組腦梗死面積明顯降低(P0.05)。HE染色,模型組見大量皺縮的細(xì)胞聚集,電針組的皺縮細(xì)胞較模型組減少。(3)缺損側(cè)大鼠大腦皮層IL-4、TNF-α蛋白表達(dá):IL-4蛋白的表達(dá),與假手術(shù)組相比,模型組和電針組蛋白表達(dá)量均明顯增多,差異均有統(tǒng)計學(xué)意義(P0.01);相比模型組,電針組蛋白表達(dá)量升高,差異有統(tǒng)計學(xué)意義(P0.05)。TNF-α蛋白表達(dá)量,與假手術(shù)組相比,模型組蛋白表達(dá)量明顯上升,差異具有統(tǒng)計學(xué)意義(P0.01),電針組蛋白表達(dá)量雖略有升高,但差異無統(tǒng)計學(xué)意義(P0.05);與模型組相比,電針組蛋白表達(dá)明顯偏低,差異具有統(tǒng)計學(xué)差異(P0.05)。IL-4/TNF-α蛋白量比值,與假手術(shù)組比較,模型組比值略有降低,但差異無統(tǒng)計學(xué)意義(P0.05),而電針組比值升高明顯(P0.01);電針組與模型組比較,差異亦有統(tǒng)計學(xué)意義(P0.01)。(4)缺損側(cè)大腦皮層IL-4mRNA、TNF-αmRNA的表達(dá)量:IL-4mRNA的表達(dá),與假手術(shù)組比較,模型組、電針組均顯著增多,差異有統(tǒng)計學(xué)意義(P0.05和P0.01);與模型組相比,電針組表達(dá)量明顯上升,差異具有統(tǒng)計學(xué)意義(P0.05)。TNF-αmRNA的表達(dá)量,相比假手術(shù)組,模型組表達(dá)量升高,差異有統(tǒng)計學(xué)意義(P0.01),電針組的差異則無明顯統(tǒng)計學(xué)意義(P0.05);與模型組相比,電針組降低,差異有統(tǒng)計學(xué)意義(P0.05)。IL-4mRNA/TNF-αmRNA的比值,與假手術(shù)組比較,模型組比值略低但無統(tǒng)計學(xué)意義(P0.05),而電針組比值升高顯著,有統(tǒng)計學(xué)意義(P0.01);電針組與模型組比較,差異亦有統(tǒng)計學(xué)意義(P0.01)。結(jié)論:電針對缺血再灌注腦損傷大鼠IL-4、TNF-α炎癥因子具有調(diào)控作用,提升抗炎細(xì)胞因子,下調(diào)促炎細(xì)胞因子,提升IL-4/TNF-α的比值,從而減輕缺血再灌注區(qū)的炎癥反應(yīng),降低梗死面積,改善神經(jīng)功能。
[Abstract]:Objective: To observe the effect of Electroacupuncture on the inflammatory response related factors TNF- alpha and IL-4 in the rat model of cerebral ischemia reperfusion, and to explore the mechanism of electroacupuncture to improve the inflammatory response in rats after cerebral ischemia, in order to further deepen the understanding of the mechanism of the treatment of ischemic encephalopathy by electroacupuncture and provide a scientific basis for clinical treatment. The SD rats were randomly selected as the sham operation group by the digital table method. The remaining animals were prepared by modified Longa thread embolus method to prepare the right cerebral artery cerebral ischemia model of the right middle cerebral artery in rats. According to the score standard of the Zea-longa5 grade 4 method, the model group and the electroacupuncture group were randomly divided into the model group and the electroacupuncture group. The effective examples of each group were randomly divided into each group. The number is 24. The sham operation group only separated the blood vessels and did not insert the thread plug; the model group used the thread plug to block the middle artery 2h after pulling out the thread to realize the reperfusion, and then did not do any treatment. The electroacupuncture group successfully selected "Baihui" and "Da Zhui" point for acupuncture, connected with the Korean HANS-200 electroacupuncture apparatus to treat 30min, the waveform was the density wave, the stimulation frequency was 2/15 Hz. after ischemia and reperfusion for 24h, the neurobehavioral score was scored by Julio's 18 score scoring system. TTC staining was used to observe the morphological changes of cerebral ischemia and the percentage of cerebral infarction area, and the pathological changes of the defect side brain tissue were observed with hematoxylin eosin (HE) staining, and the defect lateral cortex was detected by immunohistochemical staining. Tissue IL-4, TNF- alpha protein expression; changes in the expression of IL-4mRNA and TNF- alpha mRNA by qRT-PCR. Results: (1) neurological functional behavioral score: the neurological functional behavior score of rats with cerebral ischemia-reperfusion injury was significantly reduced, compared with the sham operation group (P0.01); compared with the model group, the electroacupuncture intervention could be used. The difference of neurobehavioral score was statistically significant (P0.05). (2) the morphological changes of brain tissue and the percentage of infarct area: after TTC staining, the uniform staining of the brain tissue in the sham operation group was dark red, the right cerebral ischemic brain tissue in the model group was white, the dyed area in the electroacupuncture group decreased compared with the model group. Model group and electroacupuncture group had cerebral infarction. The area percentage was higher than that of the sham operation group (P0.01 and P0.05). Compared with the model group, the area of cerebral infarction in the electroacupuncture group was significantly reduced (P0.05).HE staining. The model group saw a large number of crumpled cells aggregated, and the contraction cells in the electroacupuncture group decreased compared with the model group. (3) the IL-4, TNF- alpha protein expression of the cerebral cortex of the defected side rats: the expression of IL-4 protein and the sham operation group. The expression of protein in the model group and electroacupuncture group increased significantly, and the difference was statistically significant (P0.01). Compared with the model group, the expression of protein in the electroacupuncture group increased, and the difference was statistically significant (P0.05).TNF- alpha protein expression. Compared with the sham operation group, the amount of the model group was significantly increased, the difference was statistically significant (P0.01), the electroacupuncture group eggs were statistically significant (P0.01). Although the white expression was slightly higher, the difference was not statistically significant (P0.05). Compared with the model group, the protein expression in the electroacupuncture group was significantly lower, and the difference was statistically significant (P0.05).IL-4/TNF- alpha protein ratio. Compared with the sham operation group, the ratio of the model group decreased slightly, but the difference was not statistically significant (P0.05), but the ratio of the electroacupuncture group increased significantly (P0.01 The difference between the electroacupuncture group and the model group was statistically significant (P0.01). (4) the expression of IL-4mRNA in the cerebral cortex of the defect side and the expression of TNF- alpha mRNA: the expression of IL-4mRNA was significantly increased in the model group and electroacupuncture group compared with the sham operation group, and the difference was statistically significant (P0.05 and P0.01). Compared with the model group, the expression of the electroacupuncture group was obviously increased and the difference had the difference. The expression of.TNF- alpha mRNA in statistical significance (P0.05), compared with the sham operation group, the expression of the model group increased, the difference was statistically significant (P0.01), the difference in the electroacupuncture group was not significant (P0.05). Compared with the model group, the electroacupuncture group decreased, the difference was statistically significant (P0.05) the ratio of.IL-4mRNA/TNF- alpha mRNA, compared with the sham operation group, the model was compared with the sham group. The ratio of the group was slightly lower but not statistically significant (P0.05), but the ratio of the electroacupuncture group increased significantly (P0.01), and the difference between the electroacupuncture group and the model group was also statistically significant (P0.01). Conclusion: Electroacupuncture has a regulatory effect on IL-4, TNF- alpha and inflammatory cytokines in rats with cerebral ischemia-reperfusion injury, promoting anti-inflammatory cytokines and down-regulation of proinflammatory cells. It enhances the ratio of IL-4/TNF- alpha, thereby alleviating inflammatory reaction, reducing infarct size and improving neurological function in ischemia-reperfusion area.
【學(xué)位授予單位】：南京中醫(yī)藥大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R245

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