本文選題：嗅鞘細胞 + 人參皂甙Rg1��； 參考：《蘇州大學》2015年碩士論文

【摘要】：第一部分:人參皂甙Rg1對嗅鞘細胞遷移能力的影響目的:探討人參皂甙Rg1對嗅鞘細胞(OECs)遷移能力及遷移相關因子基質(zhì)金屬蛋白酶-2(MMP-2)、基質(zhì)金屬蛋白酶-9(MMP-9)及神經(jīng)細胞黏附分子1(NCAM1)表達的影響。方法:從新生3~5d的SD大鼠嗅球取材獲得嗅鞘細胞體外培養(yǎng)、純化后,進行人參皂甙Rg1干預(40μg/ml濃度下干預72 h)并設置對照(加等量DMEM/F12培養(yǎng)基),通過劃痕實驗和Transwell小室實驗檢測人參皂甙Rg1對嗅鞘細胞遷移能力的影響。反轉錄-聚合酶鏈反應(RT-PCR)分別測定兩組細胞遷移相關因子MMP-2、MMP-9和NCAM1的m RNA表達。通過Western-blot研究Rg1對嗅鞘細胞PI3K/Akt通路的影響。結果:Rg1組劃痕愈合面積明顯高于對照組(P0.05);Rg1組嗅鞘細胞穿過隔離膜的細胞數(shù)量較對照組明顯增多(P0.05);RT-PCR結果表明:人參皂甙Rg1顯著上調(diào)嗅鞘細胞MMP-2、MMP-9和NCAM1的m RNA表達(P0.05)。Rg1能夠促進嗅鞘細胞PI3K/Akt通路的活化。結論:人參皂甙Rg1通過上調(diào)嗅鞘細胞遷移相關因子MMP-2、MMP-9和NCAM1的表達,提高細胞的遷移能力,這一作用可能通過PI3K/Akt通路完成。第二部分:人參皂甙Rg1促進嗅鞘細胞修復脊髓損傷的實驗研究目的:研究人參皂甙Rg1對嗅鞘細胞(olfactory ensheathing cells,OECs)移植修復脊髓損傷能力的影響。方法:新生SD大鼠嗅球組織體外培養(yǎng)、純化嗅鞘細胞,人參皂甙Rg1干預并設置對照,Allen打擊建立大鼠脊髓損傷模型,隨機分成三組,對照組(control)、嗅鞘細胞移植組(OECs組)和Rg1干預嗅鞘細胞移植組(Rg1+OECs組),于移植后第1、3、7、14、28天分別進行肢體活動BBB評分,并于第28d取損傷部位脊髓組織制作石蠟切片進行免疫組化檢測;RT-PCR分別測定OECs組和Rg1+OECs組脊髓組織中CNTF和VEGF的m RNA表達。結果:Rg1+OECs組后肢運動恢復情況優(yōu)于OECs組和對照組,組織切片示Rg1+OECs組神經(jīng)元數(shù)量明顯高于OECs組和對照組(P0.01),GFAP表達量顯著低于OECs組和對照組(P0.01),HE染色示Rg1+OECs組脊髓組織結構清晰,囊性病變及細胞水腫壞死數(shù)量明顯少于其它各組;RT-PCR結果顯示人參皂甙Rg1顯著上調(diào)嗅鞘細胞在移植宿主體內(nèi)CNTF、VEGF的m RNA表達(P0.05)。結論:人參皂甙Rg1能夠通過上調(diào)嗅鞘細胞在宿主體內(nèi)CNTF、VEGF的表達,提高嗅鞘細胞移植修復脊髓損傷的能力。
[Abstract]:Part one: effects of ginsenoside Rg1 on the migration of olfactory ensheathing cells objective: to investigate the migration ability of ginsenoside Rg1 on olfactory ensheathing cells (OECs) and the migration related factors matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) and nerve cells. The effect of NCAM1) on the expression of NCAM1. Methods: olfactory ensheathing cells were cultured from olfactory bulb of SD rats for 3 days. Ginsenoside Rg1 was used to intervene at 40 渭 g/ml for 72 h. The effect of ginsenoside Rg1 on the migration of olfactory ensheathing cells was detected by scratch test and Transwell chamber experiment. Reverse transcription-polymerase chain reaction (RT PCR) was used to detect the expression of MMP-2, MMP-9 and NCAM1, respectively. The effect of Rg1 on the PI3K/Akt pathway of olfactory ensheathing cells was studied by Western-blot. Results compared with control group, the number of olfactory ensheathing cells passing through the isolation membrane was significantly higher in group 1 than that in group Rg1. The RT-PCR results showed that ginsenoside Rg1 could significantly up-regulate the expression of m RNA of MMP-2, MMP-9 and NCAM1 in olfactory ensheathing cells. It can promote the activation of PI3K/Akt pathway in olfactory ensheathing cells. Conclusion: ginsenoside Rg1 can increase the migration ability of olfactory ensheathing cells by up-regulating the expression of MMP-2, MMP-9 and NCAM1, which may be accomplished by PI3K/Akt pathway. Part two: experimental study of ginsenoside Rg1 promoting olfactory ensheathing cells to repair spinal cord injury objective: to study the effect of ginsenoside Rg1 on the ability of olfactory ensheathing cells to repair spinal cord injury. Methods: the olfactory bulb tissue of newborn SD rats was cultured in vitro, the olfactory ensheathing cells were purified, ginsenoside Rg1 was used to intervene and the control group Rg1 was used to set up the spinal cord injury model of rats, and they were randomly divided into three groups. Control group, olfactory ensheathing cell transplantation group (OECs group) and Rg1 intervention olfactory ensheathing cell transplantation group (Rg1 OECs group) were evaluated for limb activity BBB scores on day 1,1428 after transplantation. On the 28th day, paraffin sections were taken from the injured spinal cord to detect the m RNA expression of CNTF and VEGF in the spinal cord tissues of OECs group and Rg1 OECs group, respectively. Results the recovery of hind limb movement in the OECs group was better than that in the OECs group and the control group. The number of neurons in the Rg1 OECs group was significantly higher than that in the OECs group and the control group. The expression of GFAP in the Rg1 OECs group was significantly lower than that in the OECs group and the control group. The results of HE staining showed that the structure of the spinal cord in the Rg1 OECs group was clear. The results of RT-PCR showed that ginsenoside Rg1 upregulated the expression of m RNA in olfactory ensheathing cells. Conclusion: ginsenoside Rg1 can improve the ability of olfactory ensheathing cell transplantation to repair spinal cord injury by upregulating the expression of CNTF-VEGF in olfactory ensheathing cells in host.
【學位授予單位】：蘇州大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R651.2

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