淫羊藿总黄酮的药理学研究

目的 研究淫羊藿总黄酮(total flavonoids of epimedium,TFE)对人脐静脉内皮细胞损伤的保护作用.方法 将人脐静脉内皮细胞体外培养、传代后随机分为六组:空白对照组、TFE模型组、溶血性磷脂酰胆碱(LPC)损伤组、LPC+TFE(50 mg/L)组、LPC+TFE(100 mg/L) 、LPC+TFE(200mg/L)组,比较淫羊藿总黄酮对内皮细胞损伤的作用.结果 TFE模型组与空白组比较,各组数据无明显差异;LPC损伤组中MDA值、LDH值和细胞内KOS水平较空白组显著增加,而NO含量降低;在加入LPC+TFE的三组中,随着TFE的浓度增加,细胞上清液中NO含量升高,MDA和LDH含量以及ROS水平降低.结论 淫羊藿总黄酮对内皮细胞损伤具有保护作用.     

淫羊藿苷对化疗后小鼠骨髓和细胞免疫抑制作用的影响

目的:观察淫羊藿苷(ICA)对环磷酰胺(Cy)所致小鼠骨髓和免疫抑制作用的影响,探讨ICA促进化疗后小鼠造血功能和免疫功能的作用及机制。方法:将小鼠随机分为6组:正常对照组,模型组,阳性对照组,ICA高、中、低剂量组。除正常对照组小鼠外,其余各组小鼠均腹腔注射Cy(200mg/kg)。第2天开始,对ICA高、中、低剂量的实验组小鼠灌胃不同剂量的ICA(150、80、40mg/kg.d),阳性对照组小鼠尾静脉注射参芪扶正注射液(1mL/d),模型对照组给予等量生理盐水,连续给予10d。经HE染色后观察小鼠胸腺组织结构的变化。用MTT比色法检测脾淋巴细胞的增殖率。用乳酸脱氢酶(LDH)法检测腹腔巨噬细胞对肿瘤细胞的杀伤能力。用ELISA试剂盒检测混合细胞培养上清中TNF-α和IL-12的含量。用全自动血液分析仪检测外周血红细胞、白细胞和血小板数量的变化;外周血和股骨骨髓涂片经瑞氏-吉姆萨染色后,光镜下计数单根股骨骨髓细胞(BMC)的数量。结果:ICA具有保护小鼠胸腺、骨髓免受Cy损伤的作用。不同剂量的ICA组小鼠的脾淋巴细胞增殖能力增加,巨噬细胞吞噬能力和分泌细胞因子的能力均增强,外周血红细胞、白细胞和血小板数量均明显上升。结论:ICA可逆转Cy化疗后小鼠骨髓造血和免疫功能的抑制状况。     

紫外线照射对小鼠红细胞免疫粘附功能的影响

本文采用补体致敏酵母菌血凝法，检测了不同剂量紫外线照射对小鼠红细胞免疫粘附功能的影响。结果表明，０．２５ＭＥＤ剂量的紫外线照射可以提高小鼠的红细胞免疫粘附功能，２ＭＥＤ剂量的紫外线照射可使小鼠的红细胞免疫粘附功能受到抑制。实验提示，不同剂量的紫外线照射可改变机体的红细胞免疫粘附功能。     

脱水淫羊藿素对小鼠巨噬细胞免疫功能的影响

目的:研究脱水淫羊藿素(AHI)在体外对脂多糖(LPS)诱导的小鼠巨噬细胞免疫功能的影响。方法:分离制备小鼠骨髓来源巨噬细胞;CCK-8法检测不同终浓度的AHI对巨噬细胞的毒性;采用Griess试剂盒检测AHI对巨噬细胞产生NO的影响;流式细胞术(FCM)检测AHI对巨噬细胞吞噬E.coli颗粒的影响;利用FCM结合双色免疫荧光染色技术检测AHI对巨噬细胞早期活化标志CD69的表达情况;使用流式液相蛋白定量检测技术(CBA)检测AHI对LPS刺激巨噬细胞分泌细胞因子的影响。结果:终浓度为2.5、5、10μmol/L的AHI对活化的小鼠巨噬细胞均具有明显的免疫抑制作用,特别是5μmol/L AHI能明显抑制经LPS刺激的巨噬细胞早期活化,释放NO,吞噬E.coli颗粒,以及分泌IL-6、MCP-1、TNF和IL-12p70四种细胞因子。结论:AHI对LPS诱导的小鼠巨噬细胞的活化具有明显的抑制作用,是一种潜在的免疫抑制剂。     

淫羊藿甙对小鼠腹腔巨噬细胞体外增殖、吞噬和产生NO的影响

无菌分离小鼠腹腔巨噬细胞,制备单细胞悬液,加入不同终浓度的淫羊藿甙孵育4 h后,加入细菌脂多糖LPS(终浓度20 mg/L)48 h后以MTT法检测其增殖;加药孵育4 h后,分别加入直径1μm和2μm的微球(终浓度1×1010/L),5 h后利用流式细胞术(FCM)检测吞噬情况;加入LPS(终浓度20 mg/L)24 h后Griess试剂盒检测巨噬细胞NO的量。结果显示ICA在终浓度1.5、3.0μmol/L时能明显抑制LPS刺激的巨噬细胞增殖(P<0.01)和NO的产生,并促进其吞噬微球的功能。提示ICA可能抑制LPS信号转导从而抑制巨噬细胞增殖,并且通过抑制其活化,下调iNOS有关的炎症因子,使NO减少;对于未活化的巨噬细胞,ICA可能通过上调其吞噬功能,增强固有免疫系统来抵御病原体侵入。     

淫羊藿对血液透析HCV感染者细胞免疫功能的作用

为探讨丙型肝炎病毒 (HCV)感染对维持性血液透析 (血透 )患者细胞免疫功能的影响和淫羊藿对其免疫调节作用 ,应用ELISA和RT PCR检测 6 2例血透患者抗HCV和HCVRNA ,ELISA法检测患者血清sIL 2R、IL 6和TNF α水平 ,同时观察应用中药淫羊藿治疗同一血透患者 3个月前后上述指标的变化。结果表明 ,6 2例血透患者中抗HCVIgM阳性 2 7例 (4 3 6 % ) ,抗HCVIgG阳性 2 9例 (4 6 8% ) ,HCVRNA阳性 34例 (5 4 8% ) ,至少一项阳性 37例 (5 9 7% ) ;与血透患者HCV标志阴性组相比 ,阳性组血清sIL 2R、IL 6和TNF α水平显著增高 ;淫羊藿治疗后不论HCV阳性与否此三项指标均显著降低。结果提示 ,血透患者均存在明显的细胞免疫功能紊乱 ,感染HCV后免疫功能更低下 ,淫羊藿有显著调节血透患者细胞免疫功能的作用。     

氧自由基对严重烧伤后红细胞免疫粘附功能的影响

目的探讨氧自由基对烧伤后红细胞免疫粘附功能的影响。方法于伤后1d、3d、5d、7d、14d、21d、28d和35d,观察RBC-C3b受体花环形成率(RC3bRR)和RBC-IC花环形成率(RICR),以及SOD、Lpo-MDA、CAT、GSH-px和GSH水平的变化,并进行相关性分析。结果①伤后RC3bRR显著下降,RICR显著升高(P＜0.05～0.001),但4wk后逐渐恢复至正常。②伤后SOD、CAT、GSH-px和GSH的水平显著下降,Lpo-MDA的水平则显著上升(P＜0.05～0.001),至少伤后3wk才恢复到正常水平。③RC3bRR与SOD、CAT、GSH-px和GSH的水平呈显著的正相关(r值依次分别为0.601,0.799,0.715和0.597),与Lpo-MDA的水平呈显著的负相关(r=-0.564);RICR与SOD、CAT、GSH-px和GSH的水平也呈显著的负相关(r值依次分别为-0.359、-0.577、-0.420和-0.427),与Lpo-MDA的水平则呈显著的正相关(r=0.433)。结论红细胞膜极易产生脂质过氧化作用,烧伤后自由基损伤的程度,可直接影响红细胞膜的功能,特别是红细胞免疫粘附功能。在烧伤治疗过程中,提示减少机体自由基的产生,提高患者红细胞免疫粘附功能是十分必要的。     

Difference in the clustering of complement receptor type 1 (CR1) on polymorphonuclear leukocytes and erythrocytes: effect on immune adherence

Complement receptor type 1 (CR1) mediates the adherence of complement-reacted immune complexes (IC) to various blood cells. On the erythrocyte, CR1 are clustered, a distribution which favors efficient multivalent binding of C3b-coated IC. IC can also bind to CR1 expressed on polymorphonuclear (PMN) leukocytes. To evaluate the respective importance of these two cell types in immune adherence reactions, functional analysis of IC binding, as well as morphological studies of CR1 distribution at their surface were undertaken. At equal cell concentrations, resting PMN leukocytes bound the same percentage of IC as erythrocytes, despite expressing four times more CR1 at their surface. At equal CR1 concentrations, IC binding to resting or formyl-methionyl-leucyl-phenylalanine (fMLP)-stimulated PMN leukocytes was always lower than to erythrocytes. The morphological counterpart of these differences was studied by label-fracture immunoelectron microscopy. On erythrocytes, almost 50% of the CR1 were distributed in clusters of greater than or equal to 3 units, while less than 15% were grouped in such clusters on the surface of PMN leukocytes. Activation of PMN leukocytes by fMLP increased the surface density of CR1, but the proportion of clustered CR1 remained unchanged. These observations suggest that the low responsiveness of PMN leukocytes towards C3b-coated IC may be due to the unaggregated state of CR1. In the circulation, erythrocytes might function as a "buffer" for PMN leukocytes, which would otherwise engage too swiftly in reactions with IC.     

The effect of a proline-rich polypeptide (PRP) on the humoral immune response. I. Distinct effect of PRP on the T cell properties of mouse glass-nonadherent (NAT) and glass-adherent (GAT) thymocytes in thymectomized mice

It has been shown, using thymectomized, irradiated and reconstituted mice and a proline-rich polypeptide (PRP) that glass-nonadherent thymocytes contain a precursor of helper cell, and glass-adherent thymocytes a precursor of suppressor cells in the humoral immune response to SRBC. Precursor cells differentiate into helper or suppressor cells upon in vitro contact with PRP. Moreover, it was demonstrated that NAT and GAT fractions are related to PNA+ and PNA- thymocytes respectively.     

The effect of MS14 on innate and cellular immune responses in BALB/c mice

MS14 is an Iranian natural preparation of herbal-marine source with no obvious toxicity in oral administration, which possesses anti-inflammatory and immunomodulatory effects. In this study, the effect of oral administration of MS14 on nitric oxide (NO) production of peritoneal macrophages and lymphocyte Th1 cytokines and delayed-type hypersensitivity (DTH) test in BALB/c mice were investigated. Peritoneal macrophages were cultured and NO production was measured by Griess method. Viability of macrophages was assayed by MTT (3-(4,5-dimethy-2-lthiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) test. Interleukin-2 (IL-2) and INFγ levels in supernatant of spleen lymphocytes culture were assayed by enzyme-linked immunosorbent assay kit. For DTH test the mice were immunized with sheep red blood cell and DTH was measured 24?h after the last immunization of mice. NO production of macrophages has been diminished significantly in MS14 treated group (about 40%) at the presence or absence of stimulators. Macrophage viability had no significant alteration after MS14 administration. However, interferon-γ production of lymphocytes was significantly decreased in MS14 group both at the absence or presence of concanavalin A (ConA; about 50%); IL-2 production declined about 20% at the presence of ConA. In comparison with the control group, MS14 had no statistically significant effect on DTH test. The results have pointed that MS14 may have immunomodulatory potentials at least through its decreasing effect on NO production of macrophages and level of Th1 cytokine pattern.     

The effect of MS14 on innate and cellular immune responses in BALB/c mice

MS14 is an Iranian natural preparation of herbal—marine source with no obvious toxicity in oral administration, which possesses anti-inflammatory and immunomodulatory effects. In this study, the effect of oral administration of MS14 on nitric oxide (NO) production of peritoneal macrophages and lymphocyte Th1 cytokines and delayed-type hypersensitivity (DTH) test in BALB/c mice were investigated. Peritoneal macrophages were cultured and NO production was measured by Griess method. Viability of macrophages was assayed by MTT (3-(4,5-dimethy-2-lthiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) test. Interleukin-2 (IL-2) and INFγ levels in supernatant of spleen lymphocytes culture were assayed by enzyme-linked immunosorbent assay kit. For DTH test the mice were immunized with sheep red blood cell and DTH was measured 24?h after the last immunization of mice. NO production of macrophages has been diminished significantly in MS14 treated group (about 40%) at the presence or absence of stimulators. Macrophage viability had no significant alteration after MS14 administration. However, interferon-γ production of lymphocytes was significantly decreased in MS14 group both at the absence or presence of concanavalin A (ConA; about 50%); IL-2 production declined about 20% at the presence of ConA. In comparison with the control group, MS14 had no statistically significant effect on DTH test. The results have pointed that MS14 may have immunomodulatory potentials at least through its decreasing effect on NO production of macrophages and level of Th1 cytokine pattern.     

天抗(TK)对脂多糖诱导小鼠炎症模型的抗炎作用及其机制

目的研究天抗(TK)对脂多糖(LPS)诱导的小鼠炎症模型的抗炎作用及机制研究。方法将42只昆明小鼠随机分为正常对照(NC)组、模型对照(LPS)组、地塞米松(DXM)组、天抗低(TK-L)、中(TK-M)和高(TK-H)剂量组(0.2,0.8和3.2 g/kg)。各组分别灌胃给药7 d后,腹腔注射30 mg/kg的LPS诱导小鼠急性炎性模型,6 h后处死小鼠,检测小鼠脾脏指数,ELISA测定小鼠血清中IL-1β、IL-6和TNF-α的表达水平;生化法检测小鼠血清中SOD和MDA的表达;qRT-PCR检测小鼠脾脏TLR4、MyD88、TRAF6、p65、IL-1β、IL-6和TNF-αmRNA的表达水平;Western blot检测小鼠脾脏TLR4、MyD88、TRAF6、p-p65和p65蛋白表达水平。结果与LPS组相比,TK组小鼠的脾脏指数明显降低,血清和脾脏组织中IL-1β、IL-6、TNF-α和MDA水平显著下降,SOD水平明显升高,小鼠脾脏组织的TLR4、MyD88、TRAF6和p-p65等蛋白及mRNA表达水平均明显降低。结论天抗对LPS诱导的小鼠急性炎症模型具有抗炎作用,其作用机制可能是通过TLR4/MyD88/NF-κB(p-65)信号通路抑制炎症因子的释放。     

The effect of samorin (isometamedium chloride) on Trypanosoma evansi infection in mice.

Samorin (isometamedium chloride) was effective against T. evansi in mice given in single i.p. doses of 1 to 10 mg/kg. The administration of 40 mg/kg of samorin or above caused rapid death of mice and there were severe haemorrhages, degeneration and congestion in the liver, heart and kidney. The lesions in the liver and kidney were accompanied with reduced activities of ATP-ase, 5-nucleotidase, succinic tetrazolium reductase and glucose-6-phosphatase.     

In situ study on the pathogenesis and immune reaction of equine herpesvirus type 1 (EHV-1) infections in mice.

The mouse model was used to study the pathogenesis of equine herpesvirus type 1 (EHV-1) after primary and secondary intranasal infections. Within a few hours after infection, EHV-1 was found in nasal and olfactorial epithelium and sub-epithelial cells of the respiratory mucosa, but antigen-specific immune cells were never detected. Next to the lung, EHV-1 was transmitted early and directly to the brain, both via the olfactory route and the trigeminal nerve, but traces of degenerative or inflammatory processes were not detected there. In the lung, the immune cells residing or invading the parenchyma did not contain viral DNA or proteins. The primary immune response in the lungs was an alveolar and interstitial inflammation, dominated by the sequential appearance of neutrophils and macrophages, while the number of T and B lymphocytes remained unaltered. Within 24 hr after re-infection, lymphocytes accumulated around the blood vessels, outnumbering monocytes more than twofold, without neutrophils appearing. The lymphocytes comprised of little more B than T cells and the T cells were predominantly CD8+ cells. Those and B cells infiltrated the parenchyma. These results show the route of virus distribution and demonstrate the lack of antigen-specific immune cells in the lungs of mice after primary intranasal infection with EHV-1.