本文選題：人參皂甙Rd + 小膠質(zhì)細(xì)胞��； 參考：《第四軍醫(yī)大學(xué)》2014年博士論文

【摘要】：腦卒中是一種致病機制復(fù)雜，嚴(yán)重影響人類生存和生活質(zhì)量的災(zāi)難性疾病。盡管目前在治療方面取得部分相關(guān)進展，但是研究者對于如何保護卒中相關(guān)的腦損傷知之甚少。這就導(dǎo)致具有明確神經(jīng)保護作用，能夠安全運用于臨床的藥物少之又少。因此，開發(fā)具有明確保護作用的藥物對腦卒中治療具有重要的意義。人參和三七是亞洲地區(qū)應(yīng)用廣泛的傳統(tǒng)中草物，悠久的用藥史證實用藥安全有效。近來研究逐步揭示人參皂甙(ginsenosides, GS)及其有效單體成分GSRb、GSRg、GSRd等具有不同程度的腦損傷保護作用。其中，我們課題組前期的實驗工作證實，人參皂甙Rd(ginsenosides Rd, GSRd)能夠減輕谷氨酸興奮性毒性損傷，抑制神經(jīng)元胞內(nèi)鈣超載，減少腦缺血后的氧化應(yīng)激損傷，改善線粒體功能方面，具有肯定的神經(jīng)保護作用。二期和三期的臨床試驗同樣表明GSRd治療急性缺血性腦卒中患者安全有效。更為重要的是，我們發(fā)現(xiàn)在腦缺血急性期后應(yīng)用GSRd，仍然能發(fā)揮有效的腦損傷保護作用；考慮到炎癥反應(yīng)是腦缺血后期重要的病理過程，我們推測GSRd抑制炎癥損傷可能是其發(fā)揮效應(yīng)的重要機制之一。 因此，本課題旨在研究GSRd對實驗性腦缺血后的炎性反應(yīng)的影響及其可能的作用機制；同時結(jié)合GSRd和皮質(zhì)類固醇的化學(xué)結(jié)構(gòu)十分相似的特點，將GSRd和激素類藥物的作用及不良反應(yīng)進行比較，為GSRd臨床應(yīng)用于腦卒中治療提供理論依據(jù)。 實驗一人參皂甙Rd抑制實驗性腦缺血炎性反應(yīng)的研究 目的：探討GSRd對實驗性腦缺血后的炎性反應(yīng)的影響。 方法：分別采用大鼠大腦中動脈阻塞再灌注模型、LPS炎性損傷模型和氧糖剝奪模型模擬腦缺血再灌注炎性損傷，以米諾環(huán)素、地塞米松（dexamethasone，DEX）、米非司酮（RU486）作為對照。使用TTC染色測定腦梗死體積，MTT檢測GSRd對炎性損傷神經(jīng)元存活率的影響。使用實時定量PCR技術(shù)和液相芯片技術(shù)檢測GSRd（10mg/Kg）分別對缺血后炎性細(xì)胞因子mRNA和蛋白表達(dá)的影響。 結(jié)果：（1）10mg/Kg GSRd預(yù)先給藥能夠有效減少大鼠缺血后的腦梗死體積，提高受損神經(jīng)元的存活數(shù)量。DEX和RU486對缺血后腦梗死體積不產(chǎn)生影響。（2）GSRd、DEX和米諾環(huán)素預(yù)處理能夠減少多種炎性細(xì)胞因子mRNA和蛋白的表達(dá)，從而減輕缺血后的炎性損傷，但三者的抗炎作用沒有顯著性差異。（3）RU486可以阻斷或部分阻斷GSRd和DEX對細(xì)胞因子的抑制作用。 結(jié)論：GSRd預(yù)處理能夠保護腦缺血模型下的神經(jīng)元，這種作用可能與其減輕損傷后的炎性反應(yīng)、抑制細(xì)胞因子表達(dá)密切相關(guān)。 實驗二人參皂甙Rd通過作用糖皮質(zhì)激素受體發(fā)揮抗炎效應(yīng)的研究 目的：探討GSRd作用糖皮質(zhì)激素受體發(fā)揮抗炎效應(yīng)的作用機制。 方法： OGD損傷后，使用GSRd（10μM）進行干預(yù)后，使用qRT-PCR檢測不同干預(yù)處理組NFκB p65mRNA的表達(dá)。使用Western Blot檢測糖皮質(zhì)激素受體（glucocorticoid receptor，GR）通路關(guān)鍵蛋白（GR、p-IκBα/IκBα、p-p38/p38、NFκB p65）的表達(dá)水平。 結(jié)果：（1）NFκB是一種調(diào)控炎性細(xì)胞因子表達(dá)的重要轉(zhuǎn)錄因子。GSRd預(yù)處理能夠抑制IκBα磷酸化和NFκB P65入核，從而抑制NFκB活性。（2）p38是維持炎性細(xì)胞因子穩(wěn)定表達(dá)的重要因子。GSRd預(yù)處理能夠抑制p38MAPK磷酸化，減低p-p38/p38比值，降低p38活性。（3）GR激活可同時調(diào)控其下游的NFκB和p38分子。GSRd預(yù)處理能夠促進GR轉(zhuǎn)位入核，GR抑制劑RU486可以部分阻斷GSRd對缺血后GR通路蛋白表達(dá)的調(diào)節(jié)作用。 結(jié)論：GSRd可抑制炎癥調(diào)控因子NFκB和p38的活性，，這種作用可能是與其激活GR密切相關(guān)。 實驗三人參皂甙Rd與糖皮質(zhì)激素抗炎效應(yīng)及不良反應(yīng)的對比研究 目的：糖皮質(zhì)激素的特點是抗炎作用強大和用藥不良反應(yīng)廣泛。研究對比GSRd與DEX使用后的抗炎效應(yīng)及不良反應(yīng)。 方法：分別探討GSRd和DEX對代謝、免疫和骨形成方面的影響。健康C57/B6小鼠分別注射4周10mg/Kg GSRd和2.5mg/Kg DEX，對比不同干預(yù)對小鼠血糖水平，胸腺、脾臟和體重質(zhì)量的改變；采用膠原誘導(dǎo)關(guān)節(jié)炎模型對比GSRd和DEX對關(guān)節(jié)炎癥的作用；觀察不同濃度GSRd（10nM-10M）和DEX（10nM-10M）對體外培養(yǎng)成骨細(xì)胞系MC3T3-E1的增殖和分化影響。 結(jié)果：（1）和正常組相比，GSRd組小鼠的血糖、胸腺、脾臟、體重?zé)o明顯變化；相反，DEX組小鼠的血糖升高、胸腺、脾臟和體重質(zhì)量明顯下降（p0.05）。（2）膠原誘導(dǎo)關(guān)節(jié)炎模型小鼠實驗中，GSRd和DEX均可減輕關(guān)節(jié)炎性細(xì)胞浸潤，兩者之間無顯著差異。（3）在成骨細(xì)胞培養(yǎng)實驗中，GSRd和DEX均有促成骨細(xì)胞增殖和成分化的作用，但GSRd的有效劑量范圍較廣（10nM-1M）。 結(jié)論：與DEX相比，GSRd的抗炎作用同樣強大，不會影響血糖、體重和免疫器官，不會造成骨質(zhì)疏松，用藥安全，是一種極具潛力的神經(jīng)保護藥物。
[Abstract]:The study shows that ginsenoside Rd ( GS ) and its effective monomer components GSRb , GSRg and GSRd have different degrees of brain damage protection .
Considering that inflammatory reaction is an important pathological process in the late stage of cerebral ischemia , we hypothesized that GSRd can inhibit inflammatory injury as one of the important mechanisms for its effect .

Therefore , the aim of this study was to study the effect of GSRd on inflammatory response after experimental cerebral ischemia and its possible mechanism .
Combined with the characteristics of the chemical structure of GSRd and corticosteroid , the effects of GSRd and hormone drugs and adverse reactions were compared , which provided the theoretical basis for the clinical application of GSRd in the treatment of stroke .

Experimental study on the inhibition of ginsenoside Rd on the inflammatory response of experimental cerebral ischemia

Objective : To investigate the effect of GSRd on inflammatory response after experimental cerebral ischemia .

Methods : Rat brain middle cerebral artery occlusion reperfusion model , LPS inflammatory injury model and oxygen sugar deprivation model were used to simulate cerebral ischemia / reperfusion inflammatory injury . The effects of GSRd ( 10 mg / Kg ) on the survival rate of inflammatory cytokines mRNA and protein were measured by TTC staining .

Results : ( 1 ) The pre - administration of 10 mg / kg GSRd can reduce the volume of cerebral infarction after ischemia and improve the survival of injured neurons . ( 2 ) The pretreatment of DEX and RU486 can reduce the expression of various inflammatory cytokines mRNA and protein .

Conclusion : GSRd pretreatment can protect neurons in cerebral ischemia model , which may reduce inflammatory response after injury and inhibit the expression of cytokines .

Experimental study on the anti - inflammatory effect of two - two - two - ginseng saponin Rd by action of glucocorticoid receptor

Objective : To investigate the effect of GSRd on glucocorticoid receptor on anti - inflammatory effect .

Methods : After intervention with GSRd ( 10 渭M ) , the expression of NF - 魏B p65mRNA was detected by qRT - PCR . Western Blot was used to detect the expression of NF - 魏B p65 mRNA in glucocorticoid receptor ( GR ) pathway .

Results : ( 1 ) NF - 魏B is an important transcription factor regulating the expression of inflammatory cytokines . GSRd pretreatment can inhibit the phosphorylation of NF - 魏B and NF - 魏B P65 , thereby inhibiting NF - 魏B activity . ( 2 ) p38 is an important factor to maintain the stable expression of inflammatory cytokines .

Conclusion : GSRd can inhibit the activity of NF - 魏B and p38 , which may be related to the activation of GR .

Comparative study on anti - inflammatory effects and adverse effects of ginsenoside Rd and glucocorticoid

Objective : To study the anti - inflammatory effects and adverse effects of GSRd and DEX .

Methods : The effects of GSRd and DEX on metabolism , immunity and bone formation were investigated .
Collagen - induced arthritis model was used to compare the effects of GSRd and DEX on joint inflammation .
The effects of different concentrations of GSRd ( 10 nM - 10M ) and DEX ( 10 nM - 10M ) on proliferation and differentiation of MC3T3 - E1 were investigated .

Results : ( 1 ) Compared with the normal group , the blood sugar , thymus , spleen and body weight of GSRd group mice were not significantly changed .
( 3 ) GSRd and DEX all had the effects of promoting osteoblast proliferation and differentiation during osteoblast culture experiments , but the effective dose range of GSRd was wider ( 10 nM - 1M ) .

Conclusion : Compared with DEX , GSRd has the same anti - inflammatory effect , which does not affect blood sugar , body weight and immune organs . It does not cause osteoporosis and drug safety .
【學(xué)位授予單位】：第四軍醫(yī)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2014
【分類號】：R743.3

【參考文獻】

相關(guān)期刊論文 前1條

1 關(guān)永源,關(guān)超然,賀華,孫家鈞,Edwin E DANIEL;三七皂甙對血管平滑肌上受體操縱Ca~(2+)通道的特異性作用(英文)[J];Acta Pharmacologica Sinica;1994年05期

本文編號：1914046