本文關(guān)鍵詞： 多發(fā)性硬化 實驗性自身免疫性脊髓炎 帕歌斯 神經(jīng)絲蛋白 趨化因子CCL5　出處：《南華大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的： 觀察藥物帕歌斯（Pagosid,PGS）的干預(yù)對實驗性自身免疫性腦脊髓炎（experimental autoimmune encephalomyelitis,EAE）大鼠脊髓中神經(jīng)絲蛋白（neurofilament protein，NFP）及CCL5（CC Chemokine Ligand5）表達(dá)的影響，探討其對EAE是否有神經(jīng)保護(hù)作用及可能的作用機(jī)制，為MS的臨床治療提供新思路和實驗依據(jù)。 方法： 78只成年雌性Wistar大鼠隨機(jī)分為5組：正常組（N組）、免疫誘導(dǎo)組（EAE組）、免疫誘導(dǎo)組+小劑量PGS組（LPGS組）、免疫誘導(dǎo)組+大劑量PGS組（HPGS組）、免疫誘導(dǎo)組+激素組（PAT組）。使用自制完全免疫抗原（GPSCH-CAF）誘導(dǎo)EAE模型。N組不作任何處理，，EAE組造模后不予給藥，LPGS組、HPGS組分別予以PGS20mg/kg· d、100mg/kg· d灌胃，PAT組予以醋酸潑尼松8mg/kg·d灌胃。比較各組大鼠的發(fā)病率并進(jìn)行神經(jīng)功能評分，并分別于第7天、第14天、第21天處死后取脊髓行Bielschowsky銀染色、NFP及CCL5免疫組化染色。 結(jié)果： 1.各組實驗大鼠的發(fā)病率：EAE組發(fā)病率為88.89%；PGS干預(yù)組及PAT干預(yù)組發(fā)病率較EAE組降低（P0.05）；HPGS組發(fā)病率較LPGS組低（P0.05）。 2.神經(jīng)功能評分：在免疫誘導(dǎo)后第7天、第14天、第21天分別予以評分，在第14、21天，LPGS組、HPGS組及PAT組神經(jīng)功能評分均低于EAE組；第14天HPGS組、PAT組神經(jīng)功能評分低于LPGS組（P0.05）；三個時間點來比較，7天組、21天組與14天組比較，差異有統(tǒng)計學(xué)意義（P0.05），而7天組與21天組比較沒有統(tǒng)計學(xué)意義（P0.05）。 3.組織病理學(xué)：Bielschowsky銀染色示N組軸索大多走形方向一致，結(jié)構(gòu)緊密，連續(xù)性好，粗細(xì)較均勻，其余各組都不同程度地出現(xiàn)軸索改變，如結(jié)構(gòu)紊亂，連續(xù)性破壞，空泡樣缺失、斷裂。 4.免疫組織化學(xué)染色： 4.1大鼠脊髓中NFP的表達(dá)：染色結(jié)果示五組均有NFP陽性表達(dá)，其中N組表達(dá)最多；EAE組NFP陽性表達(dá)較同期N組明顯減少，而HPGS組、LPGS組、PAT組藥物干預(yù)后，NFP陽性表達(dá)神經(jīng)元數(shù)較同期EAE組增多，且差異有統(tǒng)計學(xué)意義（P0.05）；三個時間點比較，7天組、21天組與14天組比較，差異有統(tǒng)計學(xué)意義（P0.05），而7天組與21天組比較沒有統(tǒng)計學(xué)意義（P0.05）。 4.2大鼠脊髓中CCL5的表達(dá)：染色結(jié)果示除N組外，其余四組均有CCL5陽性表達(dá)，其中EAE組CCL5陽性表達(dá)較同期其他組明顯增多；藥物干預(yù)組CCL5陽性表達(dá)量有所減少，且HPGS組、PAT組CCL5表達(dá)量較同期LPGS組為低，差異有統(tǒng)計學(xué)意義（P0.05）；三個時間點比較，7天組、21天組與14天組比較，差異有統(tǒng)計學(xué)意義（P0.05），而7天組與21天組比較沒有統(tǒng)計學(xué)意義（P0.05）。 結(jié)論： 1.帕歌斯的干預(yù)可減低EAE大鼠的發(fā)病率，改善其軸索損傷程度；其干預(yù)效應(yīng)與劑量有關(guān)。 2.帕歌斯的干預(yù)對EAE大鼠的神經(jīng)保護(hù)作用機(jī)制可能與上調(diào)NFP、下調(diào)CCL5有關(guān)。
[Abstract]:Objective: Pagosid was observed. The experimental autoimmune encephalomyelitis of experimental autoimmune encephalomyelitis was induced by PGs. Protein of neurofilaments in spinal cord of rats. To explore the neuroprotective effect of NFP and CCL5(CC Chemokine Ligand5 on EAE and its possible mechanism. To provide a new idea and experimental basis for the clinical treatment of MS. Methods: Seventy-eight adult female Wistar rats were randomly divided into 5 groups: normal group (n group), immune induction group (Wistar group) and small dose PGS group (LPGS group). High dose PGS group. The EAE model was induced by using self-made complete immune antigen (GPSCH-CAF). The EAE model was induced without any treatment. The LPGS group was given 100 mg / kg / d PGS20mg/kg 路dlg 路kg 路d. The PAT group was given prednisone acetate 8 mg / kg 路d by gastric perfusion. The incidence and neurological function of rats in each group were compared and evaluated on the 7th and 14th day respectively. The spinal cord was sacrificed on the 21st day for Bielschowsky silver staining and CCL5 immunohistochemical staining. Results: 1. The incidence rate of experimental rats in each group was 88.89. The incidence rate of PGS intervention group and PAT intervention group was lower than that of EAE group. The incidence in HPGS group was lower than that in LPGS group (P 0.05). 2. Neurological function score: on the 7th day, 14th day, 21st day after immune induction, LPGS group was divided into two groups: LPGS group on day 14 and LPGS group on day 14. The scores of nerve function in HPGS group and PAT group were lower than those in EAE group. On the 14th day, the score of nerve function in HPGS group was lower than that in LPGS group (P 0.05). There was significant difference between 21 day group and 14 day group at three time points, but there was no significant difference between 7 day group and 21 day group. 3. Histopathology: Bielschowsky silver staining showed that most of the axons in the N group had the same shape, compact structure, good continuity and uniform thickness. In other groups, axonal changes occurred to varying degrees, such as structural disorder, continuity damage, void like loss and fracture. 4. Immunohistochemical staining: 4.1 expression of NFP in spinal cord of rats: the positive expression of NFP was found in all the five groups, among which N group showed the most positive expression. The positive expression of NFP in EAE group was significantly lower than that in N group, while the number of positive neurons in HPGS group was higher than that in EAE group after drug intervention. The difference was statistically significant (P 0.05). There was significant difference between the 21 day group and the 14 day group at three time points, but there was no significant difference between the 7 day group and the 21 day group. 4.2 the expression of CCL5 in spinal cord of rats: the staining results showed that there was positive expression of CCL5 in the other four groups except group N, and the positive expression of CCL5 in group EAE was significantly higher than that in other groups in the same period. The positive expression of CCL5 in the drug intervention group was decreased, and the CCL5 expression in the HPGS group was lower than that in the LPGS group (P 0.05). There was significant difference between the 21 day group and the 14 day group at three time points, but there was no significant difference between the 7 day group and the 21 day group. Conclusion: 1. The intervention of Pagos can reduce the incidence of EAE rats and improve the degree of axonal injury, and its effect is related to the dose. 2. The neuroprotective mechanism of Pagos intervention on EAE rats may be related to upregulation of NFP and down-regulation of CCL5.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R744.51

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