Preliminary Identification of Red Pigment and Positive Correlation between the Contents of Red Pigment and Total Saponins of Panax notoginseng Fruits

[Objective] This study aimed to identify red pigment of Panax notoginseng fruits and explore the correlation between pigment content and total saponins of the fruits. [Method] The red pigment of Panax notoginseng fruits was preliminarily identi- fied with specific color reactions and UV-vis spectra, and the contents of the pigment and total saponins were determined via spectrophotometry. [Result] The red hues of the fruits were contributed by anthocyanins and/or the anthocyanidins. The contents of anthocyanins and total saponins of the fruits both decreased along with thinning of the red hues. The content difference of the anthocyanins in fruits with different red hues reached extremely significant level, but that of total saponins just reached significant level. [Conclusion] The red pigment of P. notoginseng fruits is anthocyanins which are of extremely significant positive correlation with total saponins in contents.

Effects of total saponins of Panax notoginseng on immature neuroblasts in the adult olfactory bulb following global cerebral ischemia/reperfusion

The main active components extracted from Panax notoginseng are total saponins. They have been shown to inhibit platelet aggregation, increase cerebral blood flow, improve neurological behavior, decrease infarct volume and promote proliferation and differentiation of neural stem cells in the hippocampus and lateral ventricles. However, there is a lack of studies on whether total saponins of Panax notoginsertg have potential benefits on immature neuroblasts in the olfactory bulb following ischemia and reperfusion. This study established a rat model of global cerebral ischemia and reperfusion using four-vessel occlusion. Rats were administered total sa- ponins of Panax notoginseng at 75 mg/kg intraperitoneally 30 minutes after ischemia then once a day, for either 7 or 14 days. Total saponins of Panax notoginseng enhanced the number of dou- blecortin （DCX）＋ neural progenitor ceils and increased co-localization of DCX with neuronal nuclei and phosphorylated cAMP response element-binding/DCX＋ neural progenitor cells in the olfactory bulb at 7 and 14 days post ischemia. These findings indicate that following global brain ischemia/reperfusion, total saponins of Panax notoginseng promote differentiation of DCX＋ cells expressing immature neuroblasts in the olfactory bulb and the underlying mechanism is related to the activation of the signaling pathway of cyclic adenosine monophosphate response element binding protein.

Advances in Researches of Extraction, Separation, and Purification Technologies for Total Saponins of Panax notoginseng

Total saponins of Panax notoginseng have the functions of promoting blood circulation and removing phlegm, thus they have high medicinal value. There are many different extraction methods in the extraction and separation of total saponins of P. notoginseng . The extraction methods of total saponins of P. notoginseng are mainly divided into traditional extraction methods, modern extraction methods and compound extraction methods.

Advances in Researches of Extraction Processes of Total Saponins of Pnanx Notoginseng

Traditional extraction methods of total saponins of Panax notoginseng include cold soaking method,water decoction method,alcohol reflux method,percolation method,macroporous resin adsorption method,and accelerated solvent extraction( ASE) method. Modern extraction methods include ultrasonic extraction,microwave assisted extraction,supercritical CO_2 extraction,microbial fermentation assisted extraction,neural network model optimized extraction method,and multi-stage countercurrent extraction method. This paper discussed principles of these methods and compared their advantages and disadvantages.

Responses of Photosynthetic Pigment, Phenol and Total Saponin Contents of Panax notoginseng to Supplemental UV-B under Field Conditions

[Objective] The aim of this study was to investigate the content changes and their correlations of the photosynthetic pigment,phenols,including total phenols,total flavonoids and anthocyanins,and total saponins of the one-year-old P.notoginseng plants under supplemental UV-B stress in fields.[Method] The one-year-old plants were irradiated by UV-B in field for 1 min per day,and the plants under the UV-B lamp were regarded as a circle center,achieving an annular leaf-sampling.The photosynthetic pigment,phenols and total saponins of the leaves were determined spectrophotometrically.[Result] With the increase of sampling radius,the supplemental UV-B intensity decreased significantly,the contents of chlorophyll （Chl） a,Chl b,Chl （a＋b）,carotenoid （Car） and total photosynthetic pigment （Chl＋Car） of the leaves increased extremely significantly,the Chl a/b and total phenol content （TPC） decreased extremely significantly,but the Chl （a＋b）/Car changes were not significant.The contents of total flavonoids,anthocyanins and saponins all increased due to the increasing of UV-B,displaying dose effects.The UV-B intensity was positively correlated with the Chl a/b,and negatively with the Chl a,Chl b,Chl （a＋ b）,Car and （Chl＋Car） contents; and the two of TPC,total flavonoid content （TFC）,total anthocyanin content （TAC） and total saponin content （TSC） were positively correlated,all reaching extremely significant level.The UV-B intensity was positively and significantly correlated with the total flavonoid content （TFC）,negatively and significantly with the Chl （a＋b）/Car,and positively and insignificantly with the TPC,TAC and TSC.[Conclusion] For one-year-old plants of P.notoginseng,UV-B can decrease the contents of the Chl a,Chl b,Chl （a＋b）,Car and （Chl＋Car） and increase the Chl a/b and TPC,and,furthermore,induce the increases of the TFC,TAC and TSC in a dose-dependent manner.However,UV-B can hardly change the Chl （a＋b）/Car.The supplemental UV-B of well-suited dose might be one of the effective measures to improve the TSC of P.notoginseng.

Effects of Panax notoginseng saponins on apoptosis induced by hydrogen peroxide in cultured rabbit bone marrow stromal cells via altering the oxidative stress level and down-regulating caspase-3

Objective： To investigate the effects of Panax notoginseng saponins （PNS） on hydrogen peroxide （H2O2）-induced apoptosis in cultured rabbit bone marrow stromal cells （BMSCs）. Methods： The effects of different concentrations of PNS on proliferation and early osteoblast differentiation of BMSCs were determined by the MTT assay and an alkaline phosphatase （ALP） assay. An optimal effective concentration of PNS was determined and used in subsequent experiments. The cultured BMSCs were divided into three groups： untreated control, H2O2 treated, and PNS pretreatment of H2O2 treated. The oxidative stress level was assessed by superoxide dismutase （SOD） and malondialdehyde （MDA） assays. Flow cytometry was used to determine BMSC apoptosis by staining with annexinV-FITC/propidium iodide （PI）. The activity of caspase-3 enzyme was measured by spectrofluorometry. Results： PNS （0.1g/L） significantly increased both BMSC proliferation rate and ALP activity, while it decreased the indicators of oxidative stress, caspase-3 activity, and the apoptosis rate of BMSCs induced by H2O2.. Conclusion： PNS, acting as a biological antioxidant, had a protective effect on H2O2-induced apoptosis in cultured rabbit BMSCs by decreasing oxidative stress and down-regulating caspase-3.

基线等比增减设计法优选PNS/TGCO组分对实验性气虚血瘀型冠心病的影响研究

目的:初步探索三七总皂苷(PNS)和山茱萸总苷(TGCO)配伍治疗气虚血瘀型冠心病的作用及特点。方法:采用基线等比增减设计法拟定PNS/TGCO组合配比,观察不同比例组合的有效组分对垂体后叶素所致大鼠急性心肌缺血的影响以及对正常小鼠耐缺氧时间的影响。结果:处方1、2、3及血塞通有一定改善大鼠心肌缺血程度的作用,处方4、5、6与对照组比较无显著性差异;处方3、4、5、6有一定抗小鼠缺氧的作用,处方1、2及血塞通组与对照组比较无显著性差异。结论:处方3既有一定改善大鼠心肌缺血的作用又有一定抗小鼠缺氧的作用,为PNS/TGCO治疗气虚血瘀型冠心病有效组分的最佳配比。

三七皂苷PNS-5的溶血性及生物免疫活性研究

目的:研究三七皂苷PNS-5的溶血性及生物免疫活性。方法:常规方法提取PNS,大孔树脂柱层析分离纯化得PNS-5。溶血试验测定红细胞溶血率,脾淋巴细胞增殖反应测定PNS-5的生物免疫活性。结果:PNS-5溶血性较小,其HD50大于8mg/ml;PNS-5(0.1μg/ml)显著促进了ConA和LPS诱导的小鼠脾淋巴细胞增殖反应。结论:PNS-5安全性好,能促进细胞和体液免疫。

UPLC测定PNS及血塞通注射液5个主要成分含量

目的建立三七总皂苷及血塞通注射液超高效液相色谱（UPLC）测定的方法。方法采用Waters Guardcolumn色谱柱（50mm×2．1mm，1．7μm）；流动相：【水-乙腈】进行梯度洗脱；体积流量0．5mL／min；检测波长203nm；柱温25℃；进样量2μL。结果三七总皂苷及血塞通注射液中的5个成分在0．002～0．817μg与峰面积呈良好的线性关系（r=0．9999），平均回收率为101．5％，RSD值为1．31％。结论建立的UPLC法操作简便、结果可靠、重复性好、专属性强，可用于三七总皂苷及血塞通注射液中5个主要成分的含量测定。

Effects of Organic and Medium and Trace Element Fertilizers on Yield and Quality of Panax notoginseng

[Objectives]This study aimed to investigate the effects of organic and medium and trace element fertilizers(Ca,Zn,B)on yield and quality of Panax notoginseng to provide theoretical support for rational fertilization in cultivation of P.notoginseng.[Methods]Five fertilization treatments,control(CK),organic fertilizer(OM),zinc fertilizer(ZF),boron fertilizer(BF)and lime(LF),were designed.A two-consecutive-year field plot trail was conducted.The biological traits,yield and saponin content of P.notoginseng were determined.[Results]The application of organic fertilizer had no significant effect on the biological traits of P.notoginseng.Trace element fertilizers significantly increased the scape length of P.notoginseng.Among the treatments,ZF significantly increased the single flower weight but reduced the inflorescence diameter,while the effects of BF were opposite to those of ZF;LF significantly increased the stem thickness and reduced the plant height.All treatments significantly increased the seedling rate of three-year-old P.notoginseng,and the increase in the LF group(20.49%)was the largest,followed by those in the ZF(16.80%)and OM(16.40%)groups,and the increase in the BF group(13.08%)was the smallest.Although OM,ZF and BF treatments caused the root weight of individual plants to decrease,the final yield of each treatment was higher than that of the control group,and the increases in the BF and LF groups exceeded 17%(P<0.05).The total saponin outputs of all the treatments except OM were significantly higher than that of the control group.[Conclusions]Under the conditions of this test,the supplementation of organic and medium and trace element fertilizers on the basis of conventional fertilization will help to increase the yield of P.notoginseng.However,the reduction of the total saponin output of P.notoginseng caused by organic fertilizer cannot be ignored.

Combination of Total Astragalus Extract and Total Panax Notoginseng Saponins Strengthened the Protective Effects on Brain Damage through Improving Energy Metabolism and Inhibiting Apoptosis after Cerebral Ischemia-Reperfusion in Mice

Objective： To explore the effects and molecular mechanisms of the combination between total Astragalus extract （TAE） and total Panax notoginseng saponins （TPNS） against cerebral ischemia- reperfusion injury. Methods： C57BL/6 mice were randomly divided into sham-operated group, model group, TAE （110 mg/kg） group, TPNS （115 mg/kg） group, TAE-TPNS combination group and Edaravone （4 mg/kg） group, treated for 4 days, then, cerebral ischemia-repeffusion injury was established by bilateral common carotid artery （CCA） ligation for 20 min followed by reperfusion for 1 and 24 h. Results： TPNS could increase adenosine triphosphate （ATP） level, TAE and TAE-TPNS combination increased ATP, adenosine diphosphate （ADP） contents and Na＋-K＋-ATPase activity, and the effects of TAE-TPNS combination were stronger than those of TAE or TPNS alone after reperfusion for 1 h. After reperfusion for 24 h, TAE, TPNS and TAE-TPNS combination significantly increased neurocyte survival rate and decreased the apoptosis rate as well as down-regulated the expression of phosphorylated c-June N-terminal kinasel/2 （p-JNK1/2）, cytochrome C （Cyt C）, cysteine aspartic acid-specific protease （Caspase）-9 and Caspase-3. Furthermore, the effects in TAE-TPNS combination were better than those in TAE or TPNS alone. Conclusion： The combination of TAE 110 mg/kg and TPNS 115 mg/kg could strengthen protective effects on cerebral ischemia injury, the mechanism underlying might be related to improving jointly the early energy metabolism, and relieving the delayed apoptosis via inhibiting the mitochondrial apoptosis pathway of JNK signal transduction.

Total Saponins of Panax notoginseng Activate Akt/mTOR Pathway and Exhibit Neuroprotection in vitro and in vivo against Ischemic Damage

Objective:To reveal the neuroprotective effect and the underlying mechanisms of a mixture of the main components of Panax notoginseng saponins(TSPN)on cerebral ischemia-reperfusion injury and oxygenglucose deprivation/reoxygenation(OGD/R)of cultured cortical neurons.Methods:The neuroprotective effect of TSPN was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT)assay,flow cytometry and live/dead cell assays.The morphology of dendrites was detected by immunofluorescence.Middle cerebral artery occlusion(MCAO)was developed in rats as a model of cerebral ischemia-reperfusion.The neuroprotective effect of TSPN was evaluated by neurological scoring,tail suspension test,2,3,5-triphenyltetrazolium chloride(TTC)and Nissl stainings.Western blot analysis,immunohistochemistry and immunofluorescence were used to measure the changes in the Akt/mammalian target of rapamycin(mTOR)signaling pathway.Results:MTT showed that TSPN(50,25 and 12.5μg/m L)protected cortical neurons after OGD/R treatment(P<0.01 or P<0.05).Flow cytometry and live/dead cell assays indicated that 25μg/m L TSPN decreased neuronal apoptosis(P<0.05),and immunofluorescence showed that 25μg/m L TSPN restored the dendritic morphology of damaged neurons(P<0.05).Moreover,12.5μg/m L TSPN downregulated the expression of Beclin-1,Cleaved-caspase 3 and LC3 B-Ⅱ/LC3 B-Ⅰ,and upregulated the levels of phosphorylated(p)-Akt and p-mTOR(P<0.01 or P<0.05).In the MCAO model,50μg/m L TSPN improved defective neurological behavior and reduced infarct volume(P<0.05).Moreover,the expression of Beclin-1 and LC3 B in cerebral ischemic penumbra was downregulated after 50μg/m L TSPN treatment,whereas the p-mTOR level was upregulated(P<0.05 or P<0.01).Conclusions:TSPN promoted neuronal survival and protected dendrite integrity after OGD/R and had a potential therapeutic effect by alleviating neurological deficits and reversing neuronal loss.TSPN promoted p-mTOR and inhibited Beclin-1 to alleviate ischemic damage,which may be the mechanism that underlies the neuroprotective activity of TSPN.

三七总皂苷对脑缺血再灌注后神经功能恢复的影响

目的探究三七总皂苷对脑缺血再灌注后神经功能恢复的影响。方法培养30只SPF级雄性SD大鼠,构建脑缺血再灌注模型。将大鼠分为假手术组、模型组和三七总皂苷组,每组10只大鼠。检测各组大鼠脑梗面积、脑含水量。Longa评分评价大鼠神经功能缺损情况。水迷宫实验检测大鼠神经功能。检测各组大鼠血清中氧化应激指标超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)、谷光甘肽(glutathione,GSH)含量和炎症因子指标肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-10(interleukin-10,IL-10)含量。Tunel染色检测各组大鼠海马神经元细胞凋亡情况。结果和假手术组大鼠相比,模型组大鼠脑梗面积、脑含水量、神经功能缺损评分、学习潜伏期和记忆潜伏期显著升高,SOD、GSH、IL-10含量显著降低,MDA、TNF-α、IL-1β含量显著升高,海马神经元凋亡比例显著升高(P<0.05)。和模型组大鼠相比,三七总皂苷组大鼠脑梗面积、脑含水量、神经功能缺损评分、学习潜伏期和记忆潜伏期显著降低,SOD、GSH、IL-10含量显著升高,MDA、TNF-α、IL-1β含量显著降低,海马神经元凋亡比例显著降低(P<0.05)。结论三七总皂苷可能通过抑制大鼠炎症反应和氧化应激,减少海马神经元凋亡,促进脑缺血再灌注后神经功能恢复。

三七叶抗抑郁活性提取物PnGL中皂苷类成分分析

目的建立三七叶抗抑郁活性提取物PnGL中皂苷类成分含量测定的方法。方法采用香荚醛-高氯酸比色法,分别以人参二醇及人参皂苷Rb3为对照品,测定总皂苷的含量。采用HPLC法,Ultimate AQ-C18色谱柱,甲醇-水(69∶31)为流动相,检测波长203 nm,柱温为25℃,同时测定PnGL中主要成分人参皂苷Rb1、Rb2、Rc、Rb3的含量。结果以人参二醇及人参皂苷Rb3为对照品所测定的总皂苷含量,分别为97.25%、95.76%,差异不大。HPLC检测基线平稳,人参皂苷Rb1、Rb2、Rc、Rb3分离良好,分离度1.5以上,平均加样回收率分别为99.24%(RSD=1.63%)、96.41%(RSD=1.21%)、96.81%(RSD=0.86%)、100.16%(RSD=1.84%)。结论含量测定方法准确、重复性好,可为PnGL质量控制提供依据。

三七总皂苷复合纳米囊泡对糖尿病足溃疡大鼠皮肤创面愈合及血管生成的影响

目的:探究三七总皂苷(PNS)复合纳米囊泡对糖尿病足溃疡(DFU)大鼠皮肤创面愈合及血管生成的影响。方法:取雄性SD大鼠构建DFU模型,将造模成功的27只大鼠随机分为DFU组、PNS组和PNS复合纳米囊泡(PNS-CNV)组,每组9只。另取12只血糖正常的皮肤创面大鼠作为对照组(Control组)。比较各组创面愈合率。采用酶联免疫吸附试验(ELISA)法检测大鼠血清中炎症因子白介素(IL)-6、IL-8和肿瘤坏死因子-α(TNF-α)水平,苏木精—伊红(HE)染色法检测创面组织病理学变化,免疫组化染色检测创面组织微血管密度(MVD),western blotting法检测创面组织中血管生成因子(VEGF)、神经源性基因同源蛋白(Notch1)蛋白表达。结果:造模后DFU组、PNS和PNS-CNV组大鼠空腹血糖(FBG)均高于Control组(P<0.05);药物干预7 d、14 d时,与DFU组相比,PNS组和PNS-CNV组大鼠FBG明显降低,且PNS-CNV组低于PNS组(均P<0.05)。与Control组相比,DFU组大鼠创面愈合率、创面组织CD34阳性表达、MVD及VEGF蛋白表达量均明显降低,血清IL-6、IL-8、TNF-α含量及创面组织Notch1蛋白表达量升高(均P<0.05);与DFU组相比,PNS和PNS-CNV组大鼠创面愈合率、创面组织CD34阳性表达、MVD及VEGF蛋白表达量升高,血清IL-6、IL-8、TNF-α含量及创面组织中Notch1蛋白表达量降低(均P<0.05);与PNS组相比,PNS-CNV组大鼠创面愈合率、创面组织CD34阳性表达、MVD及VEGF蛋白表达量升高,血清IL-6、IL-8、TNF-α含量及创面组织Notch1蛋白表达量降低(均P<0.05)。结论:PNS-CNV可上调DFU大鼠创面组织中VEGF表达,抑制炎症反应,促进新生血管生成,从而加速创面的愈合,且该作用优于PNS。

三七总皂苷通过p38 MAPK通路抑制内毒素诱导的小胶质细胞活化

目的:探讨三七总皂苷(PNS)通过p38丝裂原活化蛋白激酶(p38 MAPK)通路对脂多糖(LPS)诱导活化的BV2小胶质细胞中肿瘤坏死因子-α(TNF-α)表达的影响。方法:将BV2小胶质细胞分为空白对照组(Control)、LPS激活组和LPS+三七总皂苷干预组(LPS+PNS)。CCK-8法检测BV2小胶质细胞的活力,确定最适合的药物干预浓度。利用Western Blot和免疫荧光检测BV2小胶质细胞中p38 MAPK和TNF-α的表达及p38 MAPK磷酸化水平(p-p38 MAPK)变化。结果:与空白对照组相比,PNS对BV2小胶质细胞的细胞活力无显著差异,最终选定100 mg/L作为药物干预浓度。Western Blot、免疫荧光结果提示,LPS激活后,BV2小胶质细胞中TNF-α的表达显著升高,p38 MAPK磷酸化水平增加(P<0.05);PNS干预后,与LPS激活组相比,TNF-α表达显著下降,p38 MAPK磷酸化水平降低(P<0.05)。使用p38 MAPK通路抑制剂(SB203580)作用后,PNS联合SB203580组(LPS+PNS+SB203580)中,TNF-α表达及p38 MAPK磷酸化水平变化与LPS+PNS组相比无显著差异(P>0.05)。此外,p38 MAPK在各组的变化无显著性差异(P>0.05)。结论:PNS可能通过p38 MAPK通路抑制活化的BV2小胶质细胞分泌的炎性因子TNF-α的表达。

三七总皂苷超声波经皮药物导入联合综合疗法治疗冈上肌肌腱炎的临床效果

目的:观察三七总皂苷超声波经皮药物导入联合综合疗法治疗冈上肌肌腱炎的临床效果。方法:选取2022年1月-2023年9月北京积水潭医院贵州医院收治的冈上肌肌腱炎患者60例作为研究对象,采用随机数字表法分为试验组和对照组,各30例。两组均给予综合疗法,试验组接受三七总皂苷葡萄糖溶液超声波经皮药物导入治疗,对照组接受体外冲击波治疗。比较两组疼痛情况、肩关节功能。结果:治疗前,两组疼痛视觉模拟评分法评分比较,差异无统计学意义(P>0.05);治疗后,两组疼痛视觉模拟评分法评分低于治疗前,且试验组低于对照组,差异有统计学意义(P<0.001)。治疗前,两组Constant-Murley肩关节功能评分比较,差异无统计学意义(P>0.05);治疗后,两组Constant-Murley肩关节功能评分高于治疗前,且试验组高于对照组,差异有统计学意义(P<0.05)。结论:三七总皂苷超声波经皮药物导入联合综合疗法治疗冈上肌肌腱炎的临床效果较好,能够减轻疼痛,改善患者肩关节功能。

Effects of Panax Notoginseng Saponins on Homing of C-kit^+ Bone Mesenchymal Stem Cells to the Infarction Heart in Rats

Objective:To investigate the effects of panax notoginseng saponins(PNS) on homing of C-kit+ bone mesenchymal stem cells(BMSCs) to the infarction heart.Methods:The acute myocardial infraction(AMI) model was established in 140 Wistar rats,105 model rats survived after operation,and the model rats were randomly divided into five groups,21 rats in each group:Western medicine group mobilized by subcutaneous injection of human granuloctye colony stimulating factor(G-CSF) 50 μg·kg-1·d-1;sham operation group and a model group treated by subcutaneous injection of normal saline 50 μg·kg-1·d-1;Chinese medicine group mobilized by intraperitoneal injection of Xuesaitong(血塞通)(ingredients of PNS) 150 mg·kg-1·d-1;integrative medicine group mobilized by subcutaneous injection of G-CSF 50 μg·kg-1·d-1 and intraperitoneal injection of Xuesaitong 150 mg·kg-1·d-1.Except for the sham-operated group,each group was divided into three sub-groups by three time points of 1 d,7 d and 14 d.G-CSF was injected once a day for 7 d.Xuesaitong was injected once a day until the rats were killed.The flow cytometry was used for detection of C-kit + cells in the peripheral blood in different time points,and immunohistochemical method was used for detection of the changes of C-kit + cell and Ki-67+ cell numbers in the marginal zone of AMI.Results:Twenty-four hours after the operation,C-kit + cells had a slight increase in the model group compared with the sham operation group(P>0.05).The peripheral blood C-kit+ cells in the integrative group increased significantly compared with the other groups on 7 d and 14 d(all P<0.05).Meanwhile the expression of C-kit + cells and Ki-67+ cells in the marginal zone of AMI in the integrative group increased significantly compared with the Chinese medicine group,the western medicine group and the model group on 1 d,7 d and 14 d(all P<0.05),and the cells in the integrative group decreased significantly on 14 d compared with that on 7 d(P<0.05).Conclusion:PNS can cooperate with G-CSF to mobilize C-kit+ BMSCs from the marrow into the peripheral blood and promote them "homing" to the infarction heart.

三七总皂苷对短暂性前脑缺血大鼠海马区神经元的修复作用实验研究

目的:探讨三七总皂苷对短暂性前脑缺血大鼠海马区神经元的修复作用。方法:选择30只SD大鼠,随机分为三七总皂苷组、模型组、假手术组,每组10只。模型组、三七总皂苷组使用四血管闭塞建立短暂性前脑缺血大鼠动物模型。假手术组手术方式同三七总皂苷组,不做卡环夹闭、电灼永久性闭塞,仅将右侧颈总动脉、颈内动脉、颈外动脉暴露,之后逐层缝合。三七总皂苷组造模后灌胃给予50 mg/kg三七总皂苷,每天2次,每次间隔12 h,模型组、假手术组灌胃给予等量的0.5%羟甲基纤维素钠。对比三组干预后7、14、28 d的海马神经细胞凋亡、新生神经元数量,对比三组大鼠的学习记忆能力,对比三组干预后7、14、28 d时测量大鼠脑梗死体积及含水量,对比三组DCX/NeuN染色阳性的细胞数量。结果:模型组干预后7、14、28 d的海马神经细胞凋亡明显较三七总皂苷组、假手术组高(均P<0.05);三七总皂苷组干预后7、14、28 d的海马神经细胞凋亡明显较假手术组高(均P<0.05);三七总皂苷组干预后7、14、28 d的海马新生神经元明显较模型组、假手术组高(均P<0.05);干预后7、14、28 d的模型组新生神经元明显较假手术组高(均P<0.05);三七总皂苷组中,随着干预时间延长,海马神经细胞凋亡明显降低,新生神经元明显升高(均P<0.05)。模型组大鼠的潜伏期、错误次数、第1记忆错误次数、第1天学习错误次数、第5天记忆错误次数、第5天学习错误次数明显较三七总皂苷组、假手术组高(均P<0.05),三七总皂苷组大鼠的潜伏期、错误次数、第1天记忆错误次数、第1天学习错误次数、第5天记忆错误次数、第5天学习错误次数明显较假手术组高(均P<0.05)。三七总皂苷组中,随着干预时间延长,第1天记忆错误次数、第1天学习错误次数、第5天记忆错误次数、第5天学习错误次数明显降低(均P<0.05)。模型组的脑梗死体积、含水量明显较三七总皂苷组、假手术组高(均P<0.05)。三七总皂苷组的脑梗死体积、含水量明显较假手术组高(均P<0.05)。三七总皂苷组中,随着干预时间延长,脑梗死体积、含水量明显降低(均P<0.05)。三七总皂苷组的DCX/NeuN染色阳性细胞数量明显较模型组、假手术组高(均P<0.05),模型组明显较假手术组高(P<0.05)。结论:三七总皂苷可促进短暂性前脑缺血大鼠海马区神经元的修复。

三七总皂苷调控SIRT1/FOXO3a/p27通路抑制大鼠肺动脉平滑肌细胞增殖

目的探讨三七总皂苷(PNS)通过SIRT1/FOXO3a/p27通路抑制大鼠肺动脉平滑肌细胞(PASMCs)增殖的作用。方法首先将PASMCs随机分为正常(Control)组和三七总皂苷(PNS)组、野百合碱(MCT)组和野百合碱+三七总皂苷(MCT+PNS)组,CCK8检测PNS对正常细胞的安全浓度,进一步检测PNS抑制增殖的最适浓度;为研究作用机制将PASMCs分为Control组、溶剂(DMSO组)组、MCT组、MCT+PNS组、MCT+PNS+SIRT1抑制剂(MCT+PNS+EX-527)组。造模结束,Edu检测细胞增殖;免疫荧光检测SIRT1、FOXO3a表达;qPCR检测细胞SIRT1、FOXO3a、p27、PCNA的表达。结果CCK8显示:0~400 mg·L^(-1)的PNS对正常细胞无毒(P>0.05),100 mg·L^(-1)的PNS可显著抑制MCT诱导的细胞增殖(P<0.01);Edu显示:MCT组较Control组增殖增多;MCT+PNS组较MCT组增殖减少;MCT+PNS+EX-527组较MCT+PNS组增殖增多。免疫荧光和qPCR显示:MCT组较Control组,SIRT1、FOXO3a、p27表达降低,PCNA表达增高;较MCT组,MCT+PNS组SIRT1、FOXO3a、p27表达增多,PCNA表达降低;较MCT+PNS组,MCT+PNS+EX-527组SIRT1、FOXO3a、p27表达降低,PCNA表达增高。结论PNS可通过SIRT1/FOXO3a/p27通路抑制大鼠PASMCs增殖。