發(fā)布時間：2018-05-08 21:12

  本文選題：通心絡(luò) + 腦微血管內(nèi)皮細胞��； 參考：《北京中醫(yī)藥大學(xué)》2014年博士論文

【摘要】：腦血管病是導(dǎo)致人類死亡的三大疾病之一,具有發(fā)病率高、致殘率高、死亡率高和復(fù)發(fā)率高的特點。其中缺血性腦血管病(Ischemic Cerebral Vascular Disease,ICVD)在世界范圍內(nèi)約占腦血管病的55%～80%,其發(fā)病率約是出血性腦血管疾病的3倍。因此,有效地防治缺血性腦血管病一直是醫(yī)學(xué)領(lǐng)域的重要研究課題之一�，F(xiàn)代醫(yī)學(xué)對于本病的治療主要為溶栓及神經(jīng)元保護兩個方面。但迄今仍無公認(rèn)的具有顯著臨床療效的藥物,這就迫使我們在缺血性腦血管病的治療中尋找新出路。隨著人們應(yīng)用中醫(yī)藥對缺血性腦病治療的不斷探索,近年來的研究重點立足于神經(jīng)血管單元的概念,通過保護血管內(nèi)皮細胞來減輕腦組織損傷。基于中醫(yī)絡(luò)病理論研發(fā)的通心絡(luò)在基礎(chǔ)與臨床研究中都具有保護缺血區(qū)微血管完整性等作用,為腦缺血治療開辟了一個新途徑。而現(xiàn)代研究已發(fā)現(xiàn)腦微血管內(nèi)皮細胞(Brain microvascular endothelial cells,BMEC)的旁分泌功能對神經(jīng)元的存活有重要調(diào)節(jié)作用,不同腦微血管內(nèi)皮細胞條件培養(yǎng)液中的差異蛋白可能是實現(xiàn)其調(diào)節(jié)作用的物質(zhì)基礎(chǔ),如胎盤生長因子(Placenta growthfactor,PLGF)對缺氧缺糖神經(jīng)元有明顯的保護作用。本研究以PLGF為切入點,分別從整體和離體研究探討腦缺血缺氧后在通心絡(luò)作用下PLGF在腦微血管內(nèi)皮細胞和腦組織分泌表達的規(guī)律及其保護神經(jīng)元的分子生物學(xué)機制,為通心絡(luò)治療缺血性腦病提供科學(xué)依據(jù)。研究目標(biāo)本研究在"營衛(wèi)交會、由絡(luò)以通"理論指導(dǎo)下,以PLGF為切入點,觀察通心絡(luò)對實驗性大鼠局灶性腦缺血損傷的保護作用及PLGF表達的影響;探討腦缺血狀態(tài)下腦微血管內(nèi)皮細胞條件液對神經(jīng)元活性及PLGF含量的影響,進而揭示PLGF保護神經(jīng)元的分子生物學(xué)機制及內(nèi)皮細胞介導(dǎo)的腦損傷保護環(huán)節(jié),圍繞神經(jīng)血管單元探討通心絡(luò)干預(yù)腦梗死的作用機制,揭示既往提出的"缺血區(qū)微血管保護—腦梗死治療新靶點"的科學(xué)內(nèi)涵,對于指導(dǎo)治療腦血管疾病的中醫(yī)藥現(xiàn)代化研究具有重要啟示。研究方法和內(nèi)容1.線栓法建立大鼠大腦中動脈阻塞的腦缺血模型。實驗動物隨機分為假手術(shù)組、模型組、通心絡(luò)組、丁苯酞組(陽性對照藥),存活期為1d、3d。各組動物在手術(shù)清醒后、術(shù)后1d、3d及取材前分別進行神經(jīng)功能評分,分?jǐn)?shù)越高損傷越重,前后兩者比值表示神經(jīng)功能恢復(fù)情況;采用HE染色觀察腦組織的病理變化;免疫熒光技術(shù)檢測MAP-2的表達以觀察神經(jīng)元的變化,確證通心絡(luò)的療效和對神經(jīng)元的保護作用。2.動物分組及存活期同上。免疫組化法檢測腦組織中PLGF表達,免疫熒光雙標(biāo)方法(PLGF+CD31、PLGF+MAP-2)分析PLGF在腦血管內(nèi)皮細胞和神經(jīng)元的表達規(guī)律;酶聯(lián)免疫吸附法(Elisa法)檢測大鼠腦組織勻漿及血清中PLGF的含量,探索腦缺血損傷大鼠腦組織中的PLGF表達部位及細胞類型。3.原代培養(yǎng)腦微血管內(nèi)皮細胞,采用CCK-8法觀察不同濃度通心絡(luò)含藥血清在作用不同時間后對腦微血管內(nèi)皮細胞活性的影響以確定其給藥的最佳濃度和時間。建立體外腦微血管內(nèi)皮細胞氧糖剝奪模型,采用CCK-8法觀察通心絡(luò)含藥血清對體外缺血損傷內(nèi)皮細胞活性的影響;利用硝酸銀還原法檢測其上清液中一氧化氮(nitric oxide,NO)、Elisa法檢測血管性血友病因子(von Willebrand factor,vWF)、PLGF的含量,分析通心絡(luò)對腦微血管內(nèi)皮細胞功能及分泌PLGF的影響。4.原代培養(yǎng)大鼠皮層神經(jīng)元,采用氧糖剝奪法建立體外神經(jīng)元缺血模型。收集通心絡(luò)含藥血清干預(yù)下腦微血管內(nèi)皮細胞不同條件液作用于培養(yǎng)的神經(jīng)元,CCK-8法檢測神經(jīng)元存活率、WST法檢測超氧化物歧化酶(Superoxide Dismutase,SOD)活性、脂質(zhì)氧化檢測法檢測丙二醛(malondialdehyde,MDA)含量變化,以探索通心絡(luò)可能通過調(diào)節(jié)內(nèi)皮細胞的旁分泌功能保護神經(jīng)元的機制。5.原代培養(yǎng)大鼠皮層神經(jīng)元并建立體外神經(jīng)元缺血模型。在MAPK信號通路阻斷劑和PI-3K信號通路阻斷劑作用下,CCK-8法檢測PLGF干預(yù)前后神經(jīng)元活性及存活率,Tunel法檢測神經(jīng)元凋亡,免疫蛋白印記法檢測不同信號通路上相關(guān)蛋白Bax、Bcl2和p-ERK1/2的表達變化,進一步從細胞信號轉(zhuǎn)導(dǎo)通路的角度探索PLGF在缺血狀態(tài)下抑制神經(jīng)元凋亡的分子生物學(xué)機制。結(jié)果1.整體實驗部分(1)動物造模醒后各組神經(jīng)功能評分無明顯差異;給藥后,各時間點通心絡(luò)組與模型組相比評分降低;將取材前與造模醒后評分的比值進行分析,相對于1d模型組,通心絡(luò)組的兩次評分比值顯著下降。相對于3d模型組,通心絡(luò)組和陽性對照藥組的兩次評分比值顯著下降。(2)HE染色結(jié)果顯示假手術(shù)組腦組織結(jié)構(gòu)完整,細胞排列整齊。模型組缺血側(cè)腦組織皮層組織結(jié)構(gòu)紊亂,神經(jīng)元細胞核固縮、濃染,部分神經(jīng)元消失。通心絡(luò)組各時間點的腦組織病理變化均較模型組有不同程度的減輕,存活神經(jīng)元數(shù)目有所增加,丁苯酞組皮層缺血區(qū)的損傷比通心絡(luò)組嚴(yán)重。(3)假手術(shù)組腦組織MAP-2熒光強度高。模型組患側(cè)皮層MAP-2的表達明顯減少,尤以梗死區(qū)為甚,可見較大范圍的弱熒光區(qū);與模型組比較,通心絡(luò)組大鼠患側(cè)皮層的MAP-2熒光表達強度增加,丁苯酞熒光強度稍有增加。(4)Elisa結(jié)果顯示PLGF在大鼠缺血腦損傷1d時血清中含量開始顯著升高,3d含量更高,大鼠腦組織勻漿中PLGF的表達呈現(xiàn)相同的表達規(guī)律;通心絡(luò)組PLGF在血清及勻漿中的表達均明顯高于假手術(shù)組和模型組,并隨時間延長而增高。(5)免疫組化結(jié)果顯示PLGF主要表達在缺血半暗帶和梗死中心的神經(jīng)元、血管內(nèi)皮細胞,腦缺血后血管內(nèi)皮細胞表達顯著增多。雙重免疫熒光標(biāo)記法顯示正常狀態(tài)下PLGF主要表達于神經(jīng)元,而缺血狀態(tài)下血管內(nèi)皮細胞也大量表達PLGF。2.離體實驗部分(1)通心絡(luò)含藥血清干預(yù)后,10%含藥血清作用24h后BMEC活性明顯提高即為通心絡(luò)含藥血清最佳濃度及作用時間。體外擬缺血模型組內(nèi)皮細胞上清液中NO、vWF含量顯著升高,而通心絡(luò)含藥血清可明顯降低NO、vWF的水平。(2)正常內(nèi)皮細胞條件培養(yǎng)液對正常神經(jīng)元和擬缺血損傷神經(jīng)元的活性無明顯影響,但通心絡(luò)含藥血清干預(yù)的正常內(nèi)皮細胞條件培養(yǎng)液對損傷的神經(jīng)元有明顯的保護作用;擬缺血損傷的內(nèi)皮細胞條件液可使正常和損傷神經(jīng)元的活性和SOD活力降低、MDA的產(chǎn)生增加,但通心絡(luò)含藥血清干預(yù)的擬缺血內(nèi)皮細胞條件培養(yǎng)液可明顯抑制這種降低,并使MDA的產(chǎn)生減少。擬缺血模型組的條件液中PLGF表達顯著升高,經(jīng)過通心絡(luò)含藥血清干預(yù)后PLGF較正常組有明顯升高。(3)相對于正常對照組,擬缺血損傷神經(jīng)元(OGD組神經(jīng)元)的存活率顯著下降,凋亡率顯著升高,相關(guān)凋亡蛋白Bcl-2的表達明顯減低而Bax顯著升高;與模型組相比,PLGF干預(yù)后神經(jīng)元存活率明顯提高,凋亡率顯著降低,Bcl-2明顯升高而Bax顯著降低;與OGD+PLGF組比,MAPK信號通路阻斷劑PD98059組神經(jīng)元存活率明顯降低,凋亡率顯著升高,Bcl-2明顯減低而Bax顯著升高,而PI-3K信號通路阻斷劑LY294002組細胞存活率等均無明顯差異。另一方面,OGD組可明顯降低p-ERK1/2表達,PLGF干預(yù)后可提高擬缺血神經(jīng)元p-ERK1/2的表達。結(jié)論1.通心絡(luò)可以促進大鼠腦缺血后神經(jīng)功能恢復(fù)、保護神經(jīng)元及增加PLGF的表達和分泌量。正常狀態(tài)下腦組織微血管內(nèi)皮細胞僅釋放少量PLGF,相反,受到缺血刺激而被激活的血管內(nèi)皮細胞可釋放大量的PLGF。故本研究證實了通心絡(luò)對實驗性大鼠局灶性腦缺血模型具有確切可靠的保護作用,并提出這種保護作用可能與PLGF相關(guān)。2.本研究首次觀察到通心絡(luò)干預(yù)下的腦微血管內(nèi)皮細胞的旁分泌功能對擬缺血神經(jīng)元的存活及氧化應(yīng)激反應(yīng)有重要的改善作用,并發(fā)現(xiàn)通心絡(luò)干預(yù)的腦微血管內(nèi)皮細胞分泌的PLGF在受到缺血缺氧刺激后會反應(yīng)性升高。3.PLGF可以提高擬缺血神經(jīng)元的存活率,本研究首次發(fā)現(xiàn)它可降低擬缺血神經(jīng)元凋亡率及調(diào)節(jié)凋亡相關(guān)蛋白的表達,并發(fā)現(xiàn)這種作用與MAPK信號通路相關(guān)�？傊�,通心絡(luò)治療腦缺血的新靶點可能是以神經(jīng)血管單元為核心改善微血管功能以介導(dǎo)腦神經(jīng)保護的作用,而該作用可能與PLGF相關(guān),PLGF可能通過MAPK信號通路發(fā)揮保護神經(jīng)元的作用。
[Abstract]:Cerebrovascular disease is one of the three major diseases that cause human death. It has the characteristics of high incidence, high disability rate, high mortality and high recurrence rate. Among them, ischemic cerebrovascular disease (Ischemic Cerebral Vascular Disease, ICVD) accounts for about 55% to 80% of cerebrovascular disease worldwide, and its incidence is about 3 times of hemorrhagic cerebrovascular disease. Effective prevention and control of ischemic cerebrovascular disease is one of the important research topics in the medical field. The treatment of modern medicine for this disease is mainly two aspects of thrombolytic and neuron protection. However, there are still no recognized drugs with significant clinical efficacy so far, which compels us to find a new way out in the treatment of ischemic cerebrovascular disease. In recent years, people use traditional Chinese medicine to explore the treatment of ischemic encephalopathy. In recent years, the focus of the research is based on the concept of neurovascular unit and the protection of vascular endothelial cells to reduce the injury of brain tissue. It has opened up a new approach for the treatment of cerebral ischemia. The paracrine function of Brain microvascular endothelial cells (BMEC) has been found to play an important role in regulating the survival of neurons in the modern study. The differential protein in the conditioned medium of different cerebral microvascular endothelial cells may be a substance to realize its regulation. Placenta growthfactor (PLGF), such as placental growth factor (PLGF), has an obvious protective effect on anoxic glucose and glucose deficiency neurons. In this study, the secretory expression of PLGF in the cerebral microvascular endothelial cells and brain tissue and the protection of neurons under the action of Tongxinluo on cerebral ischemia and hypoxia were studied with PLGF as the breakthrough point. The molecular biological mechanism provides a scientific basis for the treatment of ischemic encephalopathy by Tongxinluo. Under the guidance of the theory of "Ying Wei rendezvous, by collaterals" theory, the protective effect of Tongxinluo on focal cerebral ischemia injury in experimental rats and the effect of PLGF on cerebral ischemia were observed under the guidance of the theory of "Wei Wei rendezvous," PLGF as the breakthrough point. The effect of cell conditioned solution on the activity of neurons and the content of PLGF, and then to reveal the molecular biological mechanism of PLGF protective neurons and the protective link of brain damage mediated by endothelial cells, and to explore the mechanism of the intervention of Tongxinluo on cerebral infarction by the neurovascular unit, and reveal the new target of "microvascular protection in ischemic area - a new target for cerebral infarction" The scientific connotation of "point" has important enlightenment for the study of the modernization of Chinese medicine for the treatment of cerebrovascular diseases. The model of cerebral ischemia in rats with middle cerebral artery occlusion was established by the method and content of 1. lines. The experimental animals were randomly divided into sham operation group, model group, Tongxinluo group and butylphthalide group (positive control medicine), the survival period was 1D, 3d. groups After the operation was sober, the nerve function was scored after the operation of 1D, 3D and material after the operation. The higher the score, the heavier the injury. The ratio of the back and back showed the recovery of nerve function. The pathological changes of the brain tissue were observed by HE staining. The expression of MAP-2 was detected by immunofluorescence technique to observe the changes of neurons, and the effect of Tongxinluo and the God was confirmed to confirm the effect of the Tongxinluo and the God. The.2. group and the survival time were equal. The expression of PLGF in the brain tissue was detected by immunohistochemistry, and the expression of PLGF in the cerebral vascular endothelial cells and neurons was analyzed by PLGF+CD31 (PLGF+MAP-2). The content of PLGF in the homogenate and serum of the rat brain was detected by enzyme linked immunosorbent assay (Elisa). The expression of PLGF in brain tissue of rats with cerebral ischemia injury and cell type.3. primary culture of cerebral microvascular endothelial cells. The effect of different concentration of Tongxinluo serum on the activity of cerebral microvascular endothelial cells after different time was observed by CCK-8 method to determine the best concentration and time of the drug administration by CCK-8 method. The cell oxygen glucose deprivation model was used to observe the effect of Tongxinluo serum on the activity of endothelial cells of ischemic injury in vitro, the nitric oxide (nitric oxide, NO) in the supernatant was detected by the silver nitrate reduction method, and the content of von Willebrand factor, vWF, PLGF was detected by Elisa method, and the effect of Tongxinluo on cerebral Microblood was analyzed. The function of endothelial cells and the effect of PLGF secretion in the primary cultured cortical neurons of.4. rats, the neuron ischemia model in vitro was established by oxygen glucose deprivation. The neuron survival rate was detected by CCK-8 method and the neuron survival rate was detected by CCK-8 method. The WST method was used to detect the superoxide dismutase. The activity of Superoxide Dismutase (SOD) and the changes in the content of malondialdehyde (MDA) were detected by the method of lipid oxidation. In order to explore the mechanism of Tongxinluo to protect the neurons by regulating the paracrine function of the endothelial cells, the primary cultured rat cortical neurons were cultured and the neuron ischemic model in vitro was established. The blocking of the MAPK signal pathway was blocked. Under the action of agent and PI-3K signaling pathway blocker, CCK-8 assay was used to detect the activity and survival rate of neurons before and after PLGF intervention, Tunel method was used to detect neuronal apoptosis. The expression of Bax, Bcl2 and p-ERK1/2 in different signal pathways was detected by immunoglobulin imprint, and PLGF was further explored in the ischemic state from the angle of cell signal transduction pathway. The molecular biological mechanism of inhibiting neuron apoptosis. Results 1. the whole experiment part (1) there was no significant difference in the score of the nerve function of each group after the animal model awakening. After the administration, the score of the Tongxinluo group was lower than the model group, and the ratio of the score before and after the model was analyzed. Compared with the 1D model group, the two times of the Tongxinluo group were evaluated. Compared with the 3D model group, the score ratio of the two times of the Tongxinluo group and the positive control group decreased significantly. (2) the results of HE staining showed that the brain tissue structure of the sham operation group was complete and the cells arranged neatly. The cortical tissue structure of the ischemic brain tissue in the model group was disorganized, the nuclei of the nerve cell nuclei were condensed, concentrated and some neurons disappeared. The pathological changes of brain tissue at each time point in the collaterals group were all less than the model group, the number of surviving neurons was increased, the damage of cortical ischemic area in the butylphthalide group was more serious than that of the Tongxinluo group. (3) the MAP-2 fluorescence intensity of the brain tissue in the sham operation group was high. The expression of MAP-2 in the lateral cortex of the model group was significantly reduced, especially in the infarct area. Compared with the model group, the intensity of MAP-2 fluorescence expression in the lateral cortex of Tongxinluo group increased and the fluorescence intensity of butylphthalide increased slightly. (4) Elisa results showed that the serum content of PLGF in the ischemic brain injury of rats increased significantly and the content of 3D was higher, and the expression of PLGF in the homogenate of the rat brain presented the same table. The expression of PLGF in the serum and homogenate of Tongxinluo group was significantly higher than that in sham operation group and model group, and increased with time. (5) the results of immunohistochemistry showed that PLGF mainly expressed in the neurons of the ischemic penumbra and infarct center, vascular endothelial cells, the expression of vascular endothelial cells after cerebral ischemia increased significantly. Double immunofluorescence The labeling method showed that under normal state, PLGF was mainly expressed in the neurons, and the vascular endothelial cells in the ischemic state also expressed a large number of PLGF.2. in vitro experiment part (1) the prognosis of the blood serum of Tongxinluo. The BMEC activity was significantly improved after the action of 24h in the drug serum, which was the best concentration and time of action of Tongxinluo serum. The content of NO and vWF in the supernatant of endothelial cells increased significantly, while Tongxinluo serum could significantly reduce the level of NO and vWF. (2) the normal endothelial cell conditioned medium had no obvious effect on the activity of normal neurons and the ischemic injured neurons, but the normal endothelial cell conditioned medium of Tongxinluo serum containing drug serum was used for the injured neurons. The endothelial cell conditioned medium for ischemic injury can reduce the activity of normal and damaged neurons and the activity of SOD, and increase the production of MDA. However, this reduction can be inhibited and the production of MDA can be reduced by the intervention of Tongxinluo serum containing serum, and the production of MDA in the quasi ischemic model group is PLG. The expression of F was significantly higher than that in the normal group. (3) compared with the normal control group, the survival rate of the ischemic injured neurons (group OGD neurons) decreased significantly, the apoptosis rate increased significantly, the expression of apoptosis protein Bcl-2 decreased and Bax significantly increased. Compared with the model group, the prognosis of PLGF was compared with the model group. The survival rate of neurons was significantly increased, the rate of apoptosis decreased significantly, the Bcl-2 increased significantly, and the Bax decreased significantly. Compared with the OGD+PLGF group, the survival rate of the neurons in the PD98059 group of the MAPK signaling pathway blocker PD98059 was significantly reduced, the apoptosis rate was significantly increased, the Bcl-2 decreased significantly, and the Bax was significantly increased, but the survival rate of the LY294002 group of PI-3K signaling blockers was not clear. On the other hand, the expression of p-ERK1/2 was obviously reduced in group OGD, and the expression of p-ERK1/2 in ischemic neurons could be improved by PLGF. Conclusion 1. Tongxinluo can promote the recovery of nerve function after cerebral ischemia in rats, protect the neurons and increase the expression and secretion of PLGF. The microvascular endothelial cells in the brain tissue can release only a small amount of PLGF in normal state. On the contrary, vascular endothelial cells stimulated by ischemia can release a large number of PLGF.. This study confirms that Tongxinluo has a definite and reliable protective effect on experimental rat focal cerebral ischemia model, and it is suggested that this protective effect may be the first time to observe the cerebral microvascular endothelium under the intervention of Tongxinluo in the PLGF related.2. study. The paracrine function of the cells has an important improvement in the survival and oxidative stress response of ischemic neurons, and it is found that the reactivity of PLGF secreted by the microvascular endothelial cells in the cerebral microvascular endothelial cells intervened by Tongxinluo can increase the survival rate of.3.PLGF after ischemia and hypoxia. This study can be found for the first time that it can reduce the pseudo ischemic neuron. The apoptosis rate of ischemic neurons and the expression of apoptosis related proteins are associated with the MAPK signaling pathway. In conclusion, the new target of Tongxinluo in the treatment of cerebral ischemia may be the role of neurovascular unit as the core to improve the function of microvascular to mediate the protection of brain nerve, which may be related to PLGF, and PLGF may pass through MAPK Signaling pathways play a role in protecting neurons.

【學(xué)位授予單位】：北京中醫(yī)藥大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2014
【分類號】：R743.3

【相似文獻】

相關(guān)期刊論文 前9條

1 王江玲;黃引平;陳香娟;趙亮;張文輝;李瑩;孟媛;;子宮動脈血流多普勒彩超檢測及血清PLGF與妊娠期高血壓疾病相關(guān)性研究[J];中國婦幼保健;2007年05期

2 蘇平平;李芳;;αPLGF與抗血管新生的研究進展[J];現(xiàn)代婦產(chǎn)科進展;2009年01期

3 岳洋;蘆勝群;劉利軍;;妊娠期高血壓疾病與PLGF和MMP-9的關(guān)系[J];中國現(xiàn)代醫(yī)生;2008年34期

4 孫呈國;王棟;劉俊;;PlGF和sVEGFR-1對慢性硬腦膜下血腫的調(diào)控作用[J];江蘇醫(yī)藥;2013年22期

5 王淳;李波;方文剛;陳譽華;;PLGF通過激活Rho/ROCK信號通路介導(dǎo)人腦微血管內(nèi)皮細胞間緊密連接開放[J];基礎(chǔ)醫(yī)學(xué)與臨床;2013年08期

6 李虹;白雪峰;;子宮動脈血流動力學(xué)變化與血清PLGF水平對妊娠結(jié)局的臨床分析[J];中國醫(yī)療前沿;2007年18期

7 陳健;楊正府;何敖林;劉瑞文;胡雨佳;范然;周沁;;胎盤生長因子PLGF在胃癌中的表達及其意義的初步研究[J];實用臨床醫(yī)藥雜志;2008年11期

8 羌建峰;曹雙林;陳威;姚登福;傅琳玲;符梅;;銀杏石榴煎治療尋常性銀屑病及其對VEGF,PlGF表達的影響[J];中國皮膚性病學(xué)雜志;2008年08期

9 ;[J];;年期

相關(guān)博士學(xué)位論文 前1條

1 蓋聰;基于PLGF探討通心絡(luò)對腦缺血后微血管內(nèi)皮介導(dǎo)神經(jīng)元保護的分子機制[D];北京中醫(yī)藥大學(xué);2014年

相關(guān)碩士學(xué)位論文 前2條

1 張萍;2型糖尿病患者血清PLGF水平與頸動脈粥樣硬化的相關(guān)性研究[D];蘇州大學(xué);2015年

2 王劍利;丹參注射液對妊娠期高血壓模型鼠胎盤sFIT-1及PLGF表達的影響研究[D];山西中醫(yī)學(xué)院;2015年

，

本文編號：1863055