帕歌斯对EAE大鼠脊髓中NFP及CCL5表达的影响

发布时间：2018-01-26 23:41

本文关键词： 多发性硬化实验性自身免疫性脊髓炎帕歌斯神经丝蛋白趋化因子CCL5　出处：《南华大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：观察药物帕歌斯（Pagosid,PGS）的干预对实验性自身免疫性脑脊髓炎（experimental autoimmune encephalomyelitis,EAE）大鼠脊髓中神经丝蛋白（neurofilament protein，NFP）及CCL5（CC Chemokine Ligand5）表达的影响，探讨其对EAE是否有神经保护作用及可能的作用机制，为MS的临床治疗提供新思路和实验依据。方法： 78只成年雌性Wistar大鼠随机分为5组：正常组（N组）、免疫诱导组（EAE组）、免疫诱导组+小剂量PGS组（LPGS组）、免疫诱导组+大剂量PGS组（HPGS组）、免疫诱导组+激素组（PAT组）。使用自制完全免疫抗原（GPSCH-CAF）诱导EAE模型。N组不作任何处理，，EAE组造模后不予给药，LPGS组、HPGS组分别予以PGS20mg/kg· d、100mg/kg· d灌胃，PAT组予以醋酸泼尼松8mg/kg·d灌胃。比较各组大鼠的发病率并进行神经功能评分，并分别于第7天、第14天、第21天处死后取脊髓行Bielschowsky银染色、NFP及CCL5免疫组化染色。结果： 1.各组实验大鼠的发病率：EAE组发病率为88.89%；PGS干预组及PAT干预组发病率较EAE组降低（P0.05）；HPGS组发病率较LPGS组低（P0.05）。 2.神经功能评分：在免疫诱导后第7天、第14天、第21天分别予以评分，在第14、21天，LPGS组、HPGS组及PAT组神经功能评分均低于EAE组；第14天HPGS组、PAT组神经功能评分低于LPGS组（P0.05）；三个时间点来比较，7天组、21天组与14天组比较，差异有统计学意义（P0.05），而7天组与21天组比较没有统计学意义（P0.05）。 3.组织病理学：Bielschowsky银染色示N组轴索大多走形方向一致，结构紧密，连续性好，粗细较均匀，其余各组都不同程度地出现轴索改变，如结构紊乱，连续性破坏，空泡样缺失、断裂。 4.免疫组织化学染色： 4.1大鼠脊髓中NFP的表达：染色结果示五组均有NFP阳性表达，其中N组表达最多；EAE组NFP阳性表达较同期N组明显减少，而HPGS组、LPGS组、PAT组药物干预后，NFP阳性表达神经元数较同期EAE组增多，且差异有统计学意义（P0.05）；三个时间点比较，7天组、21天组与14天组比较，差异有统计学意义（P0.05），而7天组与21天组比较没有统计学意义（P0.05）。 4.2大鼠脊髓中CCL5的表达：染色结果示除N组外，其余四组均有CCL5阳性表达，其中EAE组CCL5阳性表达较同期其他组明显增多；药物干预组CCL5阳性表达量有所减少，且HPGS组、PAT组CCL5表达量较同期LPGS组为低，差异有统计学意义（P0.05）；三个时间点比较，7天组、21天组与14天组比较，差异有统计学意义（P0.05），而7天组与21天组比较没有统计学意义（P0.05）。结论： 1.帕歌斯的干预可减低EAE大鼠的发病率，改善其轴索损伤程度；其干预效应与剂量有关。 2.帕歌斯的干预对EAE大鼠的神经保护作用机制可能与上调NFP、下调CCL5有关。
[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.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R744.51

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