三七皂甙抗血小板聚集作用的临床观察

目的 比较三七皂甙与阿司匹林对血小板聚集率的影响,分析二者抵抗概率,探讨二者抗血小板聚集实验室指标差异.方法 三七皂甙组与阿司匹林组各60名脑梗塞患者,分别用二磷酸腺苷(ADP),花生四烯酸(从)诱导做血小板聚集试验,检测最大血小板聚集率(MAR).各组患者均于入院当天,治疗第14天检测血小板聚集率.结果 三七皂甙组与阿司匹林组血小板聚集功能在同一诱导剂作用下,未表现出统计学差异.都存在抵抗、半抵抗现象.结论 三七皂甙对血小板聚集率的影响与阿司匹林相比无统计学意义,临床有可能作为抗血小板聚集药物应用,但也存在抵抗现象.三七皂甙抗血小板聚集的长期临床效果,与其他抗血小板聚集药物联合应用能否解决抵抗问题,还需进一步,大规模临床实验研究.     

三七总皂苷后处理减轻兔失血性休克复苏期心肌损伤

目的 探讨三七总皂苷（PNS）后处理对兔失血性休克复苏期心肌损伤的影响和机制。

方法 选择雄性新西兰兔30只,11周龄,体质量2.5～3.5 kg。随机分为五组：假手术组（S组）、失血性休克复苏组（H组）、PNS后处理组（P组）、18-β甘草酸（18-AGA）+PNS后处理组（AP组）、18-AGA组（A组）,每组6只。S组行气管插管,右侧颈总动脉和右侧股静脉置管,不做放血处理。H组、P组、AP组和A组行气管插管后,制备失血性休克复苏模型。P组于造模成功后45 min给予PNS 100 mg/kg,AP组于造模前腹腔注射18-AGA 75 mg/kg,于造模成功后45 min给予PNS 100 mg/kg,A组造模前腹腔注射18-AGA 75 mg/kg。于复苏1 h时取兔心肌组织,采用Western blot法检测线粒体Cx43含量,采用TUNEL法检测心肌细胞凋亡率,通过透射电镜观察心肌线粒体超微结构,进行线粒体超微结构评分。

结果 与S组比较,H组、AP组、A组心肌线粒体Cx43相对含量明显降低（P＜0.05）,H组、P组、AP组、A组心肌细胞凋亡率明显升高,线粒体超微结构评分明显升高（P＜0.05）。与H组比较,P组心肌线粒体Cx43相对含量明显升高,心肌细胞凋亡率明显降低（P＜0.05）,AP组、A组心肌线粒体Cx43相对含量明显降低（P＜0.05）,P组和AP组粒体超微结构评分明显降低（P＜0.05）。与P组比较,AP组、A组心肌线粒体Cx43相对含量明显降低,心肌细胞凋亡率明显升高,线粒体超微结构评分明显升高（P＜0.05）。S组心肌线粒体膜完整光滑,H组线粒体嵴断裂不清,线粒体完整性丧失,P组线粒体稍肿胀但结构相对完整,AP组线粒体肿胀明显,A组线粒体嵴断裂,线粒体完整性丧失。

结论 三七总皂苷后处理可通过上调线粒体Cx43表达减轻兔失血性休克复苏期心肌损伤。     

三七总皂甙对鼠脊髓损伤早期病理变化影响的观察

Allen氏法250g·cm致伤60只Wistar大鼠T13～L1脊髓节段。伤后30min、2h、4h、第二及第三个24h分别腹腔注射三七总皂甙（PNS）100mg／kg及50mg／kg体重；另设二甲亚砜对照组及空白对照组。伤后30min、2h、6h、24h、1周及6周各取伤区脊髓组织送光、电镜检查。结果显示PNS治疗组脊髓灰质出血坏死较轻，周围白质大部存留，髓鞘残留，大量毛细血管和少突胶质细胞增生。提示PNS可减轻脊髓损伤后的灰质坏死，调节微循环和减轻继发性病理损害，为白质存活创造条件。     

脊髓损伤早期三七总皂甙抗氧自由基作用的实验研究

Ｗｉｓｔａｒ大鼠４８只随机分三组，Ａｌｌｅｎ＇ｓ脊髓损伤（ＳＣＩ）模型２５０ｇ·ｃｍ致伤Ｔ_（１３）～Ｌ_１脊髓节段，腹腔注射三七总皂甙（ＰＮＳ），伤后３０ｍｉｎ时１００ｍｇ／ｋｇ，２ｈ及４ｈ各５０ｍｇ／ｋｇ；以二甲亚砜（ＤＭＳＯ）为抗氧自由基阳性对照药物，并设立空白对照。伤后１ｈ及４ｈ取伤区脊髓组织测定丙二醛（ＭＤＡ）及超氧化物歧化酶（ＳＯＤ），发现ＰＮＳ显著减少ＭＤＡ的生成，保护ＳＯＤ活力降低。组织形态学观察到灰质区出血坏死，髓鞘分离及线粒体水肿较轻。提示ＳＣＩ早期ＰＮＳ具有明显的抗氧自由基反应和减轻继发性损害的作用。     

三七总甙对成骨细胞增殖、分化及0PG表达影响的研究

目的 观察不同剂量的三七总甙对体外培养大鼠成骨细胞增殖、分化及OPG表达的影响，探索体外作用的机制和最佳剂量。方法 第2代培养的成骨细胞分别加入终浓度为0μg／ml、10μg／ml、50μg／ml、100μg／ml的三七总甙，观察细胞生长、钙结节形成情况，碱性磷酸酶(ALP)染色、分泌量检测，骨钙素(OCN)检测，MTT法观察成骨细胞增殖，免疫组化SP法结合阳性细胞计数法检测成骨细胞OPG表达。结果 成骨细胞在7d可铺满瓶壁，30d可形成钙结节，三七总甙可促进成骨细胞的增殖ALP、OCN的分泌，以50μg／ml时作用明显。三七总甙可促进成骨细胞OPG的表达，在50μg／ml时作用明显。结论 三七总甙可促进大鼠成骨细胞的增殖、分化，促进成骨细胞OPG的表达，以50μg／ml时作用明显。     

三七总皂甙对脊髓损伤早期保护作用的实验研究

采用７２只成年健康杂种猫，随机分为正常、对照和治疗３组，Ａｌｌｅｎ法４００ｇｃｆ制作猫急性脊髓损伤模型，观察了三七总皂甙对脊髓损伤后早期６ｈ的治疗保护作用。结果表明，治疗组与对照组比较，脊髓损伤区血流量明显增加，脂质过氧化产物丙二醛含量、水含量及离子含量的异常改变得到改善，节段性脊髓诱发电位回复较好，光、电镜检查发现治疗组脊髓损伤后伤区组织损害程度明显减轻，提示三七总皂甙对脊髓损伤早期的继发性损害具有一定的保护作用。     

三七总皂甙对大鼠肝脏低温保存再灌注期间肝细胞凋亡的影响

目的　研究三七总皂甙 (PNGS)对大鼠肝脏低温保存再灌注期间肝细胞凋亡的影响及其机制。方法　采用大鼠离体肝脏再灌注模型 (IPRL) ,用Fura 2法测定低温保存 2h后肝细胞内钙离子浓度 ;经乳酸林格氏液 (LR)低温保存 2 4h的肝脏再灌注 30min后进行肝脏功能检测、氧自由基代谢产物、肝细胞凋亡、Bcl 2蛋白表达及形态学观察。LR和DMEM液中加入不同浓度PNGS。结果　大鼠肝细胞内钙离子浓度、MDA、SOD、肝细胞凋亡及Bcl 2蛋白表达阳性率等项指标各实验组明显好于对照组 (P <0 0 1) ,PNGS对大鼠肝细胞凋亡的保护作用显示出剂量依赖性 ,在 2 0 0～ 6 0 0mg范围内随剂量增加保护作用随之增强 (P <0 0 1) ,在 80 0～ 10 0 0mg范围内虽有增强 ,但无明显差别 (P >0 0 5 )。结论　PNGS减轻了大鼠肝脏在低温保存再灌注期间肝细胞凋亡 ,可能与通过抑制钙超载、抗氧自由基损伤、提高Bcl 2蛋白表达作用有关。     

三七总皂苷对大鼠肝脏低温保存作用的实验研究

目的研究三七总皂苷(PNGS)对大鼠肝脏低温保存的影响及其作用机理。方法采用大鼠离体肝脏再灌注模型(IPRL)，用Fura-2法测定DMEM液中加入不同浓度的PNGS置低温保存2h后肝细胞内钙离子浓度；将含有不同浓度PNGS的乳酸林格氏液(LR)低温保存24h的肝脏再灌注30min后检测肝脏功能、氧自由基代谢产物、能量物质和胆汁流量并进行形态学观察。结果大鼠肝细胞内钙离子浓度、MDA、AST、ALT及LDH浓度明显低于对照组．而SOD、ATP、TAN、EC含量及胆汁引流量则高于对照组(P＜O．01)．PNGS对大鼠肝脏的保护作用显示出剂量依赖性．在200～600mg范围内随剂量增加保护作用随之增强(P＜O．01)，在800～1000mg范围内虽有增强，但差异无显著性意义(P＞O．05)。结论PNGS可通过抑制钙超载、抗氧自由基损伤、促进能量物质代谢等多种机理减轻大鼠肝脏在低温保存时的损伤。     

Effects of atorvastatin on development of peritoneal fibrosis in rats on peritoneal dialysis

Rational: Peritoneal sclerosis is one of the important complications of long-term peritoneal dialysis (PD). In this study, efficacy of atorvastatin on peritoneal histology and functions in non-uremic rats on PD was tested. Objectives: Twenty-two non-uremic Wistar albino rats were randomized into three groups: Sham (intraperitoneal saline), peritoneal dialysis (PD, intraperitoneal 3.86% dextrose containing PD solution), and treatment (TX, intraperitoneal 3.86% dextrose containing PD solution plus atorvastatin added into drinking water). At the end of a 4-week period, 1 h peritoneal equilibration test was performed. Serum lipids and certain cytokines, mediators, markers, and antioxidant enzyme activities in serum and dialysate were studied. Peritoneal thickness was measured and peritoneal inflammation, fibrosis, and vascular proliferation were scored in histological sections. Main findings: In histological examinations, inflammation, fibrosis, and vascular proliferation were significantly more frequent in PD group than Sham group and it seemed to decrease significantly when atorvastatin was used in conjunction with PD. Additionally, peritoneum was significantly thicker in PD group when compared to that of Sham and TX groups. Serum parameters did not significantly differ between groups. On the other hand, dialysate glutathione reductase (GR) activity and TGF-β were significantly lower in TX group than that of the PD group, whereas dialysate IL-6 level was higher in TX group. Principal conclusions: In our study, atorvastatin use appeared to diminish structural changes in peritoneum. Decreased expression of TGF-β in dialysate may be one of the possible underlying mechanisms.     

三七总皂苷通过抑制RAGE/MAPK信号通路减轻糖尿病性骨质疏松大鼠的炎症损伤

目的 探究三七总皂苷(PNS)调控晚期糖基化终产物受体(RAGE)/丝裂原活化蛋白激酶(MAPK)信号通路对糖尿病性骨质疏松症(DOP)大鼠炎症损伤的影响。方法 将SD大鼠分成对照组、模型组、二甲双胍(MET)组(100 mg/kg)、PNS低剂量组(PNS-L组,10 mg/kg PNS)、PNS高剂量组(PNS-H组,20 mg/kg PNS)、PNS-H+空载体质粒组(20 mg/kg PNS+15μL空载体质粒)、PNS-H+RAGE过表达组(20 mg/kg PNS+15μL RAGE过表达质粒);除对照组外,其余各组大鼠通过高糖高脂饲料喂养及腹腔注射链脲佐菌素进行DOP模型构建;造模成功后各组进行相应给药处理。采用双能X线骨密度检测仪检测大鼠股骨骨密度(BMD);血糖测试仪及酶联免疫吸附法检测空腹血糖(FBG)、血清空腹胰岛素(FINS)、抗酒石酸酸性磷酸酶(TRACP)、碱性磷酸酶(ALP)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)水平;三点弯曲实验和苏木精-伊红染色分别检测股骨生物力学及病理形态变化;蛋白印迹法检测RAGE/MAPK通路蛋白表达。结果 ...     

三七总皂苷对大鼠肝移植物细胞凋亡及Bcl-2和Caspase-3 表达的影响

目的探讨三七总皂苷(PNS)预处理对大鼠供肝缺血一再灌注损伤的保护作用及其对供肝细胞凋亡和Bcl-2及Caspase-3 mRNA表达的影响。方法雄性SD大鼠分别用作供、受体．采用Kamada’s袖套法建立原位肝移植模型，根据供肝切取前1h是否静脉注射PNS(50mg／kg)将大鼠随机分为PNS预处理组(PNS组)和生理盐水对照组(NS组)；另设假手术作对照组(SO组)。分别于供肝再灌注后2h、6h及24h处死各组动物，检测血清ALT及AST，HE切片作病理组织学检查，TUNEL法检测肝细胞凋亡，RT-PCR法检测Bcl-2及Caspas-3m R-NA的表达。结果供肝再灌注后2h、6h及24h各时点，PNS组大鼠血清ALT和AST水平及肝细胞凋亡指数(A1)均明显低于NS组(P＜O．05．P＜O．01)；6h及24h，PNS组大鼠肝组织Bcl-2mRNA的表达水平明显高于NS组(P＜O．05)；2h及6h，PNS组大鼠肝组织Caspase-3mRNA的表达水平明显低于NS组(P＜O．05)。结论PNS预处理大鼠供肝，可以有效地减轻移植肝的缺血／再灌注损伤和细胞凋亡，影响细胞凋亡调控基因Bcl-2和Caspase-3的表达。这可能为PNS抗细胞凋亡的机理之一。     

三七总苷注射液辅助治疗早期糖尿病肾病患者的疗效观察

目的观察三七总苷注射液辅助治疗早期糖尿病肾病患者的疗效。方法选择早期糖尿病肾病患者66例,随机分为治疗组和对照组,均给予饮食控制、降血脂治疗及控制血糖;治疗组则增加三七总苷注射液,15d为1疗程,共2个疗程,每个疗程间隔2d。同时观察比较治疗前后患者血、尿β2-微球蛋白(β2-MG),尿微量蛋白(Alb),尿蛋白(PRO)定量定性等指标的变化。结果治疗组总有效率(84.8%)优于对照组(63.6%)(P<0.05);此外,治疗组患者血β2-MG及尿β2-MG、Alb、PRO定量定性治疗前后均有明显下降且优于对照组;治疗过程中尚未发现副作用及严重的不良反应。结论三七总苷注射液可有效改善糖尿病肾病患者的肾功能。     

Effect of renin–angiotensin system inhibitors on prevention of peritoneal fibrosis in peritoneal dialysis patients

Aim: Long‐term peritoneal dialysis (PD) may lead to peritoneal fibrosis and ultrafiltration failure. It had been demonstrated that the renin–angiotensin system (RAS) plays a key role in the regulation of peritoneal function in rats on PD. We investigated the effects of angiotensin‐converting enzyme inhibitor (ACEI) or angiotensin receptor blocker (ARB) on long‐term PD patients. Methods: We analyzed data from 66 patients treated with PD therapy at our centre for at least 12 months retrospectively, during which time at least two peritoneal equilibration tests (PET) were performed. Thirty‐eight patients were treated with ACE/angiotensin II (AII) inhibitors (ACE/ARB group); the other 28 received none of the above drugs during the entire follow up (control group). The expression of fibronectin, transforming growth factor‐β1 (TGF‐β1), Aquaporin1 (AQP1) and vascular endothelial growth factor (VEGF) in the overnight effluent were examined by enzyme‐linked immunosorbent assay. Results: The demographic data of the two groups showed no difference during the study. No difference between the groups was found with respect to residual renal function (RRF) at the start for both groups by the end of follow up, decreased in the vast majority of patients from both groups (P = 0.014). After 12 months, a significant difference in ultrafiltration was found between the two groups: in the control group it had decreased, while it had not changed in the ACE/ARB group (P < 0.05). In comparison with the baseline level, expression of fibronectin, TGF‐β1 and VEGF in dialysate effluent were significantly increased except for AQP1 in the control group (P < 0.05), but not in the ACE/ARB group (P > 0.05). Conclusion: The findings suggest that ACE/AII inhibitors appeared to have a slower rate of decline in ultrafiltration and RRF, effectively protect against peritoneal fibrosis in long‐term peritoneal dialysis. Long‐term follow up seems to be required to draw more conclusions.     

Inhibition of Rho-kinase alleviates peritoneal fibrosis and angiogenesis in a rat model of peritoneal dialysis

Abstract

Background/Aims: The present study investigated whether Rho-kinase inhibition had a therapeutic role on the pathogenesis of peritoneal fibrosis and angiogenesis. Methods: A rat model of peritoneal dialysis was induced by a daily intraperitoneal infusion of 4.25% Dianeal. Those rats were treated with Rho-kinase inhibitor, fasudil. Immunofluorescence, Western blot and RT-PCR were used to detect the expression of TGF-β1, Collagen I, αSMA and VEGF in each group. Microvessel density (MVD) was measured by immunohistochemistry. Rho-kinase activity was determined by western immunoblotting. Results: Rho-kinase was activated in the peritoneum of the PD group, which was inhibited by fasudil. Compared with PD group, the mRNA and protein expressions of TGF-β1, αSMA and Collagen I were significantly downregulated in fasudil treatment groups in a dose-dependent manner, and the expression of VEGF and peritoneal MVD was also significantly downregulated in fasudil treatment groups in a dose-dependent manner. Conclusion: The Rho-kinase was activated in the peritoneum of the peritoneal dialysis rats, and the inhibition of Rho-kinase by fasudil can remarkably decrease peritoneal fibrosis and angiogenesis.     

Longitudinal changes in peritoneal kinetics: the effects of peritoneal dialysis and peritonitis

BACKGROUND.: Peritoneal infection and poor ultrafiltration continue to bethe major causes of treatment failure in CAPD. The combinedeffects of peritonitis and the continuous exposure to dialysisfluid remain the most likely candidates affecting the peritoneumin the long term. The purpose of this study was to observe theeffects of peritonitis and dialysis on longitudinal peritonealfunction. METHODS.: The peritoneal equilibration test (PET) was utilized to quantifylongitudinal changes in low-molecular-weight solute transfer(D/P_creat) and ultrafiltration (UF) in 233 patients treatedwith CAPD. Of these, 166 represented an unselected cohort (Group1) studied prospectively from commencing treatment for up to54 months, and 67 were selected patients (Group 2) with PETdata available at commencement of the study, having been ondialysis for a minimum of 18 months. PETs were performed either6-monthly or following peritonitis episodes. RESULTS.: Data on the short-term effect of peritonitis kinetics were pooledfor groups 1 and 2. Single, isolated episodes (n = 86) had nosignificant effect on D/P_creat or UF, whereas recurrences orclusters of infection (n = 70) caused increases in D/Pcreatand reductions in UF, the significance of which increased withthe number of episodes. There were significant correlationsbetween both changes in D/P_creat and UF with the cumulativedialysate leukocyte count, regardless of infecting organism,suggesting that intensity of peritoneal inflammation is alsoimportant. Those organisms associated with greater change inperitoneal kinetics, e.g. S. aureus, Pseudomonas, also had thehighest neutrophil counts. The longitudinal changes in peritoneal kinetics were analysedfor patients in group 1 only. There was a highly significantincrease in D/P_creat after 6 months treatment; this increasedfurther with time on treatment, reaching further significanceat 42 and 48 months. There was an associated reduction in UF.In view of the short-term effects of peritonitis on kineticsgroup 1 was further subdivided into patients who were eitherperitonitis free or only experienced isolated infections, group1a, and those that had multiple infection episodes, group 1b.Treatment drop-out, due to death or technical failure occurredat double the rate in group 1b, who also had significantly higherD/P_creat and lower UF at 1, 6, 12, 18 and 24 months of treatment.Group 1a subsequently caught up, however, indicating that peritonitisis not the only factor influencing long-term changes in peritonealkinetics. CONCLUSIONS.: These data suggest that solute transfer increases and UF declineswith time on peritoneal dialysis. This process is exacerbatedand accelerated by peritonitis, and appears to be proportionalto the degree of associated inflammation and number of infectionsin close proximity.     

Role of transforming growth factor beta in peritoneal fibrosis

SUMMARY: Technique survival of peritoneal dialysis is seriously limited by the development of peritoneal fibrosis. the mesothelial cell layer lining the peritoneum is important in the pathogenesis of peritoneal fibrosis. Mesothelial cells are able to produce transforming growth factor beta (TGF-β), and respond to stimulation by this cytokine. In this review, we will detail the evidence available so far for the role of the complex interaction between TGF-β and mesothelial cells in the development of peritoneal fibrosis.