从补肾生血药对衰老小鼠IL—2、IL—2R的效应探讨肾生髓与免疫调节的关系

目的探讨肾生髓与免疫调节的关系.方法采用老年小鼠和D-半乳糖致衰老小鼠两种衰老肾虚动物模型,分别连续灌服补肾生血药两个月和1个月,应用MTT法检测脾淋巴细胞增殖,放射免疫技术测定IL-2的含量,流式细胞技术分析L-2R的表达情况.结果老年小鼠和D-半乳糖致衰老小鼠脾淋巴细胞增殖能力、IL-2含量、IL-2R表达明显低于青年小鼠和正常对照小鼠(P＜0.001),给药后则可明显得到提高(P＜0.05～0.01).结论肾生髓与免疫调节密切相关,补肾生血药能增强机体的免疫功能.     

枸杞多糖对衰老小鼠原癌基因c—myc表达的影响

目的：探讨枸杞多糖抗衰老的分子机理。方法：采用老年小鼠和D－半乳糖致衰老小鼠两种衰老动物模型,分别连续灌服枸杞多糖2个月和1个月,应用半定量逆转录－聚合酶链式反应技术（RT－PCR）检测小鼠骨髓原癌基因c-myc表达的变化。结果：老年小鼠和D－半乳糖致衰老小鼠骨髓原癌基因c-myc表达是枸杞多糖抗衰老分子机理之一（P＜0．001）,给药后则可明显得到抑制（P＜0．05）。结论：抑制原癌基因c-myc表达是枸杞多糖抗衰老分子机理之一。     

补肾生血解毒方对再生障碍性贫血小鼠白细胞生成的影响

目的：探讨补肾生血解毒方对理化因素致骨髓抑制造成再生障碍性贫血（AA）模型小鼠白细胞生成的影响。方法：BALB／C小鼠随机分为空白组、模型组和补肾生血解毒方小、中、大剂量组,除空白组以外其余各组小鼠以60Co一γ射线复合环磷酰胺致小鼠骨髓造血抑制,补肾生血解毒方小、中、大剂量组分别给予不同剂量中药灌胃,其余2组予等量生理盐水灌胃。血细胞计数仪检测小鼠外周血中白细胞（WBC）的改变,酶联免疫吸附测定（ELISA）检测外周血血清中粒细胞一巨噬细胞集落刺激因子（GM—CSF）含量的差异,real—timePCR检测骨髓细胞中GM—CSF受体仪、B基因表达的改变。结果：与模型组比较,补肾生血解毒方可以明显提高AA模型小鼠外周血白细胞数量（小剂量作用14d）,显著增加外周血血清中GM—CSF含量和骨髓细胞中GM—CSF受体基因的表达。结论：补肾生血解毒方治疗AA发挥疗效作用．可能与提高GM—CSF活性和诱导造血细胞分化为成熟的白细胞．并促进其增殖、发挥免疫调节作用有关。     

补肾生血药对衰老小鼠Bcl-2mRNA表达的影响

“肾生骨髓”是中医肾生理功能的重要组成部分。因此,肾生髓理论的现代研究在肾本质研究中占有重要的地位。我们实验室自20世纪70年代末以来一直致力于肾生髓理论的现代研究,研究的重要成果之一是补肾生血药的研制成功。该药由熟地黄、何首乌、山茱萸、枸杞子、阿胶等11味中药组成,具有滋阴补肾,益髓生血等功能。20多年来,我们通过对补肾生血药的药效机理的研究,来阐述中医经典理论肾生髓的现代内涵。基因是决定生命结构和功能的最根本物质,与     

补髓生血冲剂对小鼠造血功能的实验研究

为了探讨补髓生血冲剂对再生障碍性贫血患的药物作用机理，对失血性贫血模型小鼠及环磷酰胺（CY）导致骨髓抑制模型小鼠进行治疗观察。对小鼠外周血WBC，RBC，Hb，网织红细胞（RC）及骨髓有核细胞（BMC），脾脏重量和多能造血干细胞（CFU－S）等指标进行检测，并与目前临床上常用的省优部优产品再障生血片进行了对比，以探讨补髓生血冲剂对小鼠造血系统的影响。结果表明补髓生血冲剂具有明显促进骨髓造血和脾脏髓外造血的作用，并且对升高WBC、RC、BMC及CFU－S等方面明显优于再障生血片。     

补肾生血和补肾调经方药对老龄雌性金黄地鼠生殖器官组织形态的影响

补肾生血复方中药由生地、熟地、龟板胶、山萸肉等１１味中药组成，具有补肾生髓、造血补血、滋阴填精补髓作用［１、２］，研究表明肾生髓理论的分子基础具有提高ＤＮＡ含量、ＤＮＡ聚合酶活性、ＤＮＡ甲基化水平、血红蛋白ｍＲＮＡ转录水平的作用［３、４］。补肾调经复...     

中国神方补肾生血药抗衰老分子机理的研究

本文就补肾生血药作用的分子基础作了深入研究。结果表明，补肾生血药能明显提高老龄大鼠ＤＮＡ合成代谢水平，提高骨髓有核细胞内ＤＮＡ、核酸含量，明显提高ＤＮＡ聚合酶活力，该药还能提高鼠肝ＤＮＡ甲基化酶活力及ＤＮＡ甲基化水平     

百合多糖抗氧化作用研究

百合多糖200、400mg/kg ig,可使D－半乳糖致衰老小鼠血中SOD、CAT及GSH－Px活力升高,血浆、脑匀浆和肝匀浆中LPO水平明显下降。     

益髓生血颗粒对不同发育阶段小鼠α-血红蛋白稳定蛋白表达的影响

目的：探讨益髓生血颗粒对不同发育阶段小鼠α-血红蛋白稳定蛋白(alpha-hemoglobin stabilizing protein,AHSP)表达的影响。方法：分别提取正常成年小鼠骨髓、孕鼠(孕21 d)和胎鼠(孕14 d)胎肝胎脾有核细胞总RNA,采用real-time RT-PCR方法检测中药益髓生血颗粒干预后对不同发育阶段小鼠的α-珠蛋白稳定蛋白基因表达的影响。结果：中药干预后,胎鼠(孕14 d)、孕鼠(孕21 d)的AHSP基因表达无明显改变,而成年鼠骨髓AHSP基因表达明显上调。结论：益髓生血颗粒治疗β-地中海贫血的可能分子机制之一,是通过促进AHSP基因的表达,减少游离 α-珠蛋白的沉积,遏止其对红细胞的毒害作用,减少溶血发生而达到治疗效果。     

益髓生血颗粒对辐射损伤小鼠骨髓细胞周期的影响

目的:观察补肾生血药益髓生血颗粒对骨髓细胞周期的影响,探讨其可能的造血调控机制。方法:将78只雄性昆明种小鼠随机分为正常组(A组)、模型组(B组)、G-CSF组(C组)、益髓生血颗粒高剂量组(D组)、益髓生血颗粒中剂量组(E组)、益髓生血颗粒低剂量组(F组),每组13只。采用60Co-γ射线全身一次性照射造成小鼠骨髓急性损伤模型。造模前,先预防给药2 d,D-F组分别按12,6,3 g.kg-1.d-1灌胃益髓生血颗粒,A～C组给予等量蒸馏水。造模第2天,C组按30μg.kg-1.d-1皮下注射G-CSF注射液,其余各组给药方法同预防给药,连续14 d。收集各组小鼠骨髓有核细胞,采用流式细胞仪检测骨髓细胞周期的分布情况。结果:与正常组相比,模型组G0/G1期细胞比例为(67.6±7.1)%,明显降低;S期细胞比例为(25.3±5.7)%,G2/M期细胞比例为(7.1±1.8)%,均明显增加(均P<0.001);细胞增殖指数为(32.4±7.1)%,明显升高(P<0.001)。与模型组相比,益髓生血颗粒高剂量组G0/G1期细胞比例为(62.2±3.6)%,明显降低(P<0.05);细胞增殖指数为(37.8±3.6)%,明显升高(P<0.05);益髓生血颗粒各剂量组处于S期的细胞比例分别为(29.1±2.7)%,(29.2±4.8)%,(29.5±3.9)%,均明显升高(均P<0.05)。结论:补肾生血药益髓生血颗粒可通过促进骨髓细胞由G0/G1期进入S期而明显促进细胞增殖。     

基于“肾生髓、髓生血”理论治疗地中海贫血

地中海贫血是因先天性基因缺陷致使珠蛋白合成障碍引起的一种遗传性血液病,也叫珠蛋白合成障碍性贫血。它是世界上发病率高、危害最大的单基因遗传病,至今尚无有效的治疗方法。中国中医科学院广安门医院根据肾藏精生髓、髓生血理论,在广西高发区,用补肾益髓法治疗地中海贫血600余例,取得了肯定疗效。论文探讨了中药治疗地中海贫血的证治规律,总结了近30年的临床实践,提出了中医治疗地中海贫血的理论基础和中医核心病机假说,形成了系统的理论与有效治法。依托多项国家基金,从整体效应、基因突变与疗效关系、骨髓有效造血、红细胞结构与功能、调控珠蛋白mRNA表达等不同层面,探讨了中药治疗地中海贫血疗效特点和作用靶点的分子机制,使中药治疗地中海贫血整体水平达到新的高度。研究结果将从一个侧面揭示"肾藏精"及"与肾相关"疾病的内在关系。从"肾"论治地中海贫血的临床与基础研究成果,为阐释"肾藏精"脏象理论的科学内涵提供了有说服力的证据。     

双黄升白颗粒剂升高白细胞作用机制的研究

目的:观察双黄升白颗粒(黄芪、黄精、女贞子、天花粉、骨碎补等)对白细胞减少症模型小鼠骨髓细胞增殖及粒单系集落刺激因子(GM-CSF)生成的影响,并探讨其作用机制。方法:Se-137放射源照射造成小鼠白细胞减少症动物模型,采用MTT比色法测定小鼠骨髓细胞增殖程度:采用双抗体夹心Elisa法测定小鼠血清中GM-CSF含量;采用SP免疫细胞化学方法测定培养的小鼠脾细胞胞浆中GM-CSF蛋白表达。结果:双黄升白颗粒能够促进模型小鼠骨髓细胞增殖,并能升高模型小鼠血清GM-CSF含量,促进小鼠脾细胞GM-CSF蛋白表达。结论:双黄升白颗粒能够促进白细胞减少症模型小鼠骨髓细胞增殖,升高其血清GM-CSF含量,并促进小鼠脾细胞GM-CSF蛋白表达,这可能是其具有升高白细胞作用的内在机制之一。     

“髓”为中心的治疗靶点

干细胞及其组织微环境是"髓"本质的生物学基础,围绕"髓"形成单个或多个"靶点"的集合就是"髓"为中心的治疗靶点。针对"髓"的单个或多个靶点的不同组合可提出若干个不同治疗方案,包括基于"补肾生髓成肝"的肝硬化、肝衰竭、肝癌防治方案及"补肾生髓"防治艾滋病免疫重建不全的方案,极大地提高了中医药防治肝藏病及其相关病证的能力和水平。"髓"是"肝肾同源"的中心环节,极大地深化和提高了"肝肾同源"的理论认识,明确了深入研究中医药调控再生修复的作用及其机制的重点研究方向。     

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??OBJECTIVE To study the blood enriching effect and mechanism of Jiuweiejiao ointment on mouse model of blood deficiency syndrome.METHODS Blood deficiency syndrome model was established with cyclophosphamide(CTX) and acetylphenylhydrazine(APH). Total of 60 ICR mice were divided into normal group, model group, Siwu granule group, Jiuweiejiao ointment high-dose, medium-dose and low-dose group (15.6, 7.8, 3.9 g??kg^-1). Each group mouse were treated with the respective drug through intragastrical administration once a day for consecutive 14 d. The levels of red blood cells (RBC), leukocytes (WBC), hemoglobin (HGB) level in blood of mice were detected by blood analyzer; the histopathological changes of bone marrow were determined by HE staining; the levels of erythropoietin (EPO), interleukin-3 (IL-3), tumor necrosis factor-?? (TNF-??), granulocyte colony stimulating factor (G-CSF) and granulocyte-macrophage colony stimulating factor (GM-CSF) in serum were measured by enzyme linked immunosorbent assay (ELISA) method; the expression level of EPO protein in kidney tissue, G-CSF and GM-CSF protein in bone marrow tissue were detected by Western-blot method.RESULTS Compared with model group, Jiuweiejiao ointment could significantly increase the levels of RBC, WBC and HGB in blood of mice, markedly improve bone marrow hyperplasia, reduce the non-hematopoietic tissue and adipocytes, obviously repaire the hematopoietic tissue structure; significantly decrease the level of TNF-?? and increase the level of EPO, IL-3, G-CSF and GM-CSF in serum. In addition, Jiuweiejiao ointment could also obviously increase the expression of EPO protein in kidney tissue and G-CSF, GM-CSF protein in bone marrow tissue compared with that in model group.CONCLUSION Jiuweiejiao ointment has obvious blood enriching effects, which may be related with promoting the secretion of hematopoietic growth factors and reducing the inhibition of hematopoietic function by negative hematopoietic factor.     

中医“肾髓系统”刍议

肾藏精,精生髓,髓充脑、养骨、生血,"肾、精、髓"物质与功能共同构成了中医"肾髓系统",其中"肾为本,精为要,髓为枢",肾中精气盛衰决定髓的盈亏,是脑为髓海、肾主骨、精化血的生理和病理基础;同时,中医现代研究发现干细胞与肾精关系密切,体现了"肾髓系统"的物质和功能特点。     

Studies on pharmacological junctions of hairy root of Astragalus membranaceus]

OBJECTIVE: To investigate the effect of hairy root of Astragalus membranaceus(HRA). METHOD: HRA were given 10 g/kg per day for 50 days to aged mice treated with D-galactose, and the effect on memory and antioxidant functions were estimated. After administration of HRA 10 g/kg for four days, anti-ischemia-reperfusion kidney model of rat was prepared. The kidney function, activity of superoxide dismutase (SOD) and content of malondialdehyde(MDA) in kidney were examined. Mice with immunosuppression induced by cyclophosphamide were given orally HRA 10 g/kg for 12 days. The activity of natural killer (NK) cells was measured. RESULT: HRA improved the memory, raised SOD activity in brain and liver, decreased the MDA content in the liver of aged mice, reduced the MDA content in ischemia-reperfusion kidney, decreased the creatinine level in blood of rats, and promoted the activity of NK cells in immunosuppressed mice. CONCLUSION: Similar to the natural A. membranaceus, HRA has senility-preventing, antioxidizing and immunomodulating functions.     

从整体论到系统生物学进行肾虚和衰老的研究

中医学对世界医学的贡献之一就是整体观,我们从整体观出发研究了肾阳虚证与下丘脑－垂体－靶腺轴功能紊乱的联系;采用基因芯片技术研究老年大鼠（肾虚证）及中药干预后下丘脑－垂体－肾上腺－胸腺（HPAT）轴的分子网络调控规律;通过基因芯片也揭示存在于老年大鼠下丘脑、垂体、肾上腺、淋巴细胞、骨骼、肝脏、肾脏等与“肾”相关组织的神经－内分泌－免疫以及神经－内分泌－骨代谢两大基因网络路线,从而推进“证”的整体性研究。近年按照系统生物学的动态、干预、多层面整合的要求,在全基因组转录水平、代谢组和单个衰老相关信号通路3个不同层面揭示衰老在不同层面的共同规律;揭示出基因的时空关联随着衰老进程逐渐弱化;并且中药淫羊藿总黄酮能有效改善衰老在上述两方面的系统改变。因此从整体观到系统生物学的转变可能促进中医学进入现代生命科学领域。     

Hematopoietic effect of fractions from the enzyme-digested colla corii asini on mice with 5-fluorouracil induced anemia

Effects of fractions A and B from enzyme-digested traditional Chinese medicine colla corii asini on mice with 5-fluorouracil-induced anemia were investigated. The purpose of this study was to further understand the hematopoietic activities and mechanisms of colla corii asini. The fractions A and B were administered to anemic mice for 12 days. After confirming the anti-anemic effect of fractions A and B, we examined the effects of fractions A and B on immature granulocyte and erythroid cell activity and plasma cytokine level. Fraction A administration at 2 g/kg and 1 g/kg and fraction B administration at 1.6 g/kg and 0.8 g/kg activated granulocyte and erythrocyte progenitor cells in bone marrow and erythrocyte progenitors in spleen, led to the recovery of white blood cell and red blood cell counts, and increased the percentage of peripheral reticulocytes in red cells. The GM-CSF and EPO production determined by examining GM-CSF mRNA and EPO mRNA in the kidney and liver of the anemic mice were also enhanced. This treatment significantly increased serum GM-CSF and EPO level and lowered serum transforming growth factor (TGF-beta) level. These results suggested that fractions A and B promoted hematopoiesis by activating immature granulocyte and erythroid cells, partly by stimulating GM-CSF and EPO secretion and suppressing TGF-beta release. Identification of a specific peptide or protein is still required for the development of a novel medicine for anemia caused by malignancy or chemotherapy.     

李佑生主任医师从“胆”论治肾性骨营养不良

肾性骨营养不良是慢性肾脏病引起的钙、磷代谢紊乱的临床综合征,以骨痛、骨质疏松、骨折等病理性改变为主要表现。李佑生教授受《黄帝内经》“胆主骨所生病”理论启发,将“胆主骨”与“肾主骨”理论相结合,根据肾性骨营养不良的临床特点,以及“他脏治本脏病”的理念,从胆主骨理论入手治疗肾性骨营养不良。肾性骨营养不良的病机为肾虚髓空,胆腑郁滞,加之脾气亏虚,肝宗筋束骨不利,瘀血、浊毒壅滞经脉,以肾虚、胆郁为本,三焦闭阻、瘀血浊毒内蕴为标。据此立补肾、调胆两大治则,并以补肾温髓调胆为纲,健脾补肾、补益肝肾、温髓壮胆化瘀、疏胆升阳祛浊为法,从整体观入手,辨证施治,达到阴阳协调,五脏六腑和安。以期在临床为他脏治本脏病、本脏病同治提供理论依据。