Research progress of Lycium barbarum polysaccha-ride in preventing and treating Alzheimer's disease / 中国药理学与毒理学杂志

Alzheimer's disease(AD)is a neurode-generative disease with insidious onset and progressive development.In recent years,the prevalence of AD has shown a linear upward trend.At present,its pathogene-sis is not clear.Lycium barbarum polysaccharide(LBP)is one of the main effective components extracted from the dried ripe fruit of Lycium barbarum L.,a solanaceae plant.It has many pharmacological effects such as anti-aging,anti-oxidation,anti-fibrosis,anti-inflammation,neu-roprotection and immunomodulation.LBP has been widely studied in the field of prevention and treatment of AD because of its good anti-aging and neuroprotective effects.Its prevention and treatment mechanism mainly includes the following points:① Regulating the apoptosis of nerve cells.Studies have shown that the signal pathway com-posed of phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt)participates in a series of processes such as the growth,proliferation and apoptosis of neurons and plays an important regulatory role.LBP can reduce the number of cell apoptosis,increase the expression levels of autophagy protein Beclin1 and microtubule-associated protein 1 light chain 3Ⅱ(LC3Ⅱ),and decrease the expres-sion levels of p-Akt and phosphorylated mammalian target protein of rapamycin(p-mTOR),which indicates that Lycium barbarum polysaccharide can prevent and treat AD by inhibiting PI3K/Akt/mTOR pathway and improv-ing the autophagy level of cells.②Inhibition of amyloid β-protein(Aβ)production.Aβ is the main component of senile plaque,which is regarded as the main biomarker of AD.It is found that the neurotoxicity of Aβ plays a role by increas-ing the influx of Ca2+ mediated by N-methyl-D-aspartate receptor in the process of signal transduction in the brain,and then generating reactive oxygen species(ROS)and apoptosis signals.LBP can promote autophagy of HT22 cells by inhibiting PI3K/Akt pathway,which has a protec-tive effect on Aβ-induced toxicity.③ Inhibit the produc-tion of inflammatory cytokines.In the pathogenesis of AD,microglia are activated when they feel pathological accumulation of Aβ,and then cell surface immune and adhesion molecules such as cluster of differentiation 45(CD45),CD40,CD36 and integrins are activated,thereby recruiting Src family kinases and activating MAPK path-way,leading to over-expression of proinflammatory fac-tors.A large number of cytokines and chemokines are produced,which may lead to synapse damage and loss.For example tumor necrosis factor-α(TNF-α)can induce neuronal apoptosis and injury.The production of interleu-kin,and other cytokines and chemokines may also lead to microglia activation,astrocyte proliferation,and further secretion of proinflammatory factors and amyloid deposi-tion,thus making the neuroinflammatory cascade perma-nent.LBP can down-regulate the expression of TNF-α and IL-1β genes,reduce the level of intracellular ROS,and improve the learning and memory ability of AD patients.In this paper,the mechanism of Lycium barbarum polysaccharide in preventing and treating AD is reviewed,in order to provide basis for drug development and clini-cal application.

Inhibition of photoreceptor apoptosis in mice with retinitis pigmentosa through NF-κB/NLRP3 pathway suppression with Lycium barbarum polysaccharide / 数字中医药（英文）

Objective@# To explore whether Lycium barbarum polysaccharide (LBP) can reduce the apoptosis of retinal photoreceptor cells in retinitis pigmentosa (RP) mice by inhibiting nuclear factor-kappa B (NF-κB)/NOD-like receptor thermal protein domain-associated protein 3 (NLRP3) signaling pathway. @*Methods@# (i) In vitro experiments, mouse retinal ganglion cells (661W cells) were divided into normal, model, LBP low-dose (LBP-L, 40 mg/L), LBP middle-dose (LBP-M, 80 mg/L), LBP high-dose (LBP-H, 160 mg/L), and positive drug control (NLRP3 inhibitor, 160 mg/L) groups. And the 661W cells were exposed to varying concentrations of H2O2 ranging from 50 to 400 μmol/L to determine the optimal concentration for inducing apoptosis (200 μmol/L). Then the cell viability was assessed using Cell Counting Kit-8 (CCK-8), while the apoptosis rate was detected by flow cytometry; the expression of NLRP3 was detected by immunofluorescence; and the expression of apoptosis markers was detected by enzyme-linked immunosorbent assay (ELISA) and Western blot (WB). (ii) In vivo assays were carried out with the use of C57/BL6 and Rd10 mice. The animal experimental groups were divided into normal, model, LBP-L, LBP-M, LBP-H, and NLRP3 inhibitor groups, in which the normal group was C57/BL6 mice and the other groups were Rd10 mice. Ten mice were included in each group, and the corresponding drugs were administered intragastrically for a duration of four weeks. NF-κB/NLRP3 pathway and the expression of apoptosis markers were observed by electroretinogram, histopathological examination, and WB to assess the effects of LBP on retinal photoreceptor cell apoptosis.@*Results@#(i) In vitro experiments, compared with the normal group, the apoptosis rate of 661W cells in model group was significantly increased (P < 0.01), and the expression levels of key proteins of NF-κB/NLRP pathway, such as NLRP3, NF-κB, p-NF-κB, and pro-apoptotic protein caspase-3, were up-regulated (P < 0.01). The rate of Bax/Bcl-2 was increased (P < 0.01), and the concentrations of interleukin (IL)-1β and tumor necrosis factor (TNF)-α were significantly increased (P < 0.01). Compared with the model group, high dose of LBP decreased the apoptosis rate of 661W cells (P < 0.01), and down-regulated the expression levelsof the key proteins of NF-κB/NLRP3 pathway, including NF-κB, NLRP3, p-NF-κB, and caspase-3 (P < 0.01). The rate of Bax/Bcl-2 was decreased (P < 0.01), and the concentrations of IL-1β and TNF-α were decreased (P < 0.01). (ii) In vivo experiments, high dose of LBP significantly increased morphological changes in the outer nuclear layer (ONL) thickness of Rd10 mice, as well as functional changes in the amplitudes of the a-wave and b-wave (P < 0.01), which also down-regulated the expression levels of NF-κB (P < 0.05), NLRP3, p-NF-κB, and caspase-3 (P < 0.01), reduced the Bax/Bcl-2 rate (P < 0.01), and decreased the concentrations of IL-1β (P < 0.01) and TNF-α (P < 0.05). @*Conclusion@#LBP could improve both retinal morphology and function, providing protection to photoreceptors from apoptosis through the inhibition of the NF-κB/NLRP3 pathway.

Screening of active components and preliminary mechanism exploration of Lycii Fructus for improving osteoporosis based on the zebrafish model / 药学学报

Juvenile zebrafish were used to screen the active components of Lycii Fructus for improving osteoporosis. The screening results were further verified by zebrafish adult osteoporosis model and the action mechanism was explored. Prednisolone was used as the inducer to build osteoporosis models of juvenile and adult zebrafish, and 9 groups of samples of different extracts and chemical parts of Lycii Fructus were given. Alizarin red staining was applied for observing the scale matrix mineralization and bone resorption. The activities of osteoblasts and osteoclasts were detected using alkaline phosphatase (ALP) and tartrate resistant acid phosphatase (TRAP/TRACP) staining. The expressions of bone metabolism-related genes alp, osteoprotectin (opn), osteoblast specific transcription factor (sp7), cathepsin K (ctsk), tracp, and Runt family transcription factor 2b (runx2b) in each group were determined using quantitative polymerase chain reaction. The results showed that all components of Lycii Fructus improved the formation area of the first vertebrae, the staining light density value, and the number of vertebrae joints in juvenile zebrafish and the Lycium barbarum polysaccharide (LBP) treatment group exerted the best effect. In addition, LBP prevented the formation of bone resorption lacunae in zebrafish scales, increased ALP activity, decreased TRAP activity, up-regulated the alp, sp7, and opn genes, and lowered the expressions of ctsk and tracp genes. In conclusion, LBP regulated the activity of osteoblasts and osteoclasts, reduced bone resorption, promoted bone formation and enhanced bone density, which might be the main anti-osteoporosis active fraction of Lycii Fructus. This study provided modern scientific evidence for the scientific connotation of the traditional effect of "strengthening bones and muscles" of Lycii Fructus, provided the reference for the evaluation of the anti-osteoporosis activity of traditional Chinese medicine based on zebrafish adult model, and provided beneficial enlightenment for the bone health needs of the aging society population.

Lycium barbarum Polysaccharide Regulates Activation of RAW264.7 Macrophages Through MGL/TLR Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the mechanism of Lycium barbarum polysaccharides （LBP） in promoting the activation of RAW264.7 macrophages. MethodRAW264.7 macrophages were stimulated with LBP at different concentrations （50， 100， 200 mg·L-1）， and those stimulated with lipopolysaccharide （LPS） at 100 μg·L-1 and galactose （Gal） at 100 mg·L-1 as positive controls. After 24 h of LBP stimulation， the cell counting kit-8 （CCK-8） was used to detect the survival rate of RAW264.7 macrophages treated with LBP （0， 50， 100， 200， 400， 800 mg·L-1）. The levels of interleukin-6 （IL-6） and interleukin-12 （IL-12） in cell culture supernatant were detected by enzyme-linked immunosorbent assay （ELISA）. The protein and mRNA expression of p38 mitogen-activated protein kinase （MAPK）， extracellular signal-regulated kinase （ERK）， c-Jun N-terminal kinase （JNK）， and nuclear factor κB （NF-κB） in Toll-like receptor 4 （TLR4）/Toll-like receptor 2 （TLR2）/macrophage galactose-type lectin （MGL） pathway of RAW264.7 macrophages was detected by Real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR） and Western blot. ResultCCK-8 results showed that compared with the results in the blank group， the survival rate of RAW264.7 macrophages decreased in the 400， 800 mg·L-1 LBP groups （P<0.05）. ELISA results showed that compared with the blank group， 50 mg·L-1 LBP could promote the secretion of IL-12 in RAW264.7 macrophages （P<0.01）. Compared with the blank group， 100 mg·L-1 LBP and 200 mg·L-1 LBP could promote the secretion of IL-6 in RAW264.7 macrophages （P<0.05， P<0.01）. Western blot results showed that compared with the blank group， the LBP groups （50， 100， 200 mg·L-1） enhanced protein expression levels of MAPK key molecules （p-p38 MAPK， p-ERK， p-NF-κB， and p-JNK） in TLR4， TLR2， and MGL pathways （P<0.05， P<0.01）. Compared with the model group， the 200 mg·L-1 LBP group could promote the expression level of p-NF-κB protein in RAW264.7 macrophages （P<0.01）. Real-time PCR results showed that compared with the blank group， the LBP groups （50， 100， and 200 mg·L-1） enhanced the mRNA expression levels of MAPK key molecules （p38 MAPK， ERK， NF-κB， and JNK） in TLR4 and TLR2 pathways （P<0.05， P<0.01）. Compared with the model group， the 50 and 200 mg·L-1 LBP groups could promote the mRNA expression levels of JNK and ERK2 in RAW264.7 macrophages （P<0.05， P<0.01）. ConclusionLBP can regulate the activation of RAW264.7 macrophages and participate in the immune response through the TLR2/TLR4/MGL pathway.

A novel anticancer property of Lycium barbarum polysaccharide in triggering ferroptosis of breast cancer cells / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Breast cancer is one of the most malignant tumors and is associated with high mortality rates among women. Lycium barbarum polysaccharide (LBP) is an extract from the fruits of the traditional Chinese herb, L. barbarum. LBP is a promising anticancer drug, due to its high activity and low toxicity. Although it has anticancer properties, its mechanisms of action have not been fully established. Ferroptosis, which is a novel anticancer strategy, is a cell death mechanism that relies on iron-dependent lipid reactive oxygen species (ROS) accumulation. In this study, human breast cancer cells (Michigan Cancer Foundation-7 (MCF-7) and MD Anderson-Metastatic Breast-231 (MDA-MB-231)) were treated with LBP. LBP inhibited their viability and proliferation in association with high levels of ferroptosis. Therefore, we aimed to ascertain whether LBP reduced cell viability through ferroptosis. We found that the structure and function of mitochondria, lipid peroxidation, and expression of solute carrier family 7 member 11 (SLC7A11, also known as xCT, the light-chain subunit of cystine/glutamate antiporter system Xc-) and glutathione peroxidase 4 (GPX4) were altered by LBP. Moreover, the ferroptosis inhibitor, Ferrostatin-1 (Fer-1), rescued LBP-induced ferroptosis-associated events including reduced cell viability and glutathione (GSH) production, accumulation of intracellular free divalent iron ions and malondialdehyde (MDA), and down-regulation of the expression of xCT and GPX4. Erastin (xCT inhibitor) and RSL3 (GPX4 inhibitor) inhibited the expression of xCT and GPX4, respectively, which was lower after the co-treatment of LBP with Erastin and RSL3. These results suggest that LBP effectively prevents breast cancer cell proliferation and promotes ferroptosis via the xCT/GPX4 pathway. Therefore, LBP exhibits novel anticancer properties by triggering ferroptosis, and may be a potential therapeutic option for breast cancer.

Effect and relative mechanism of Lycium Barbarum Polysaccharide on LPS-induced inflammatory response within human retinal pigment epithelial cells / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To explore the effect of lycium barbarum polysaccharides(LBP)on inflammatory response of human retinal pigment epithelial cells(ARPE-19)induced by lipopolysaccharide(LPS)and its possible signal pathway.<p>METHODS: ARPE-19 cells were stimulated by LPS <i>in vitro</i> to construct the inflammatory injury cell model. Primarily, the cells were divided into five groups randomly. The blank group was cultured in complete medium, and the LPS group was stimulated with complete medium containing 10μg/mL LPS for 24h. The low, medium and high concentration LBP groups were incubated with complete medium importing 0.1, 0.5 and 1mg/mL LBP for 24h separately, and then stimulated with complete medium containing 10μg/mL LPS for 24h. We used the CCK-8 method to observe the cell survival rate, real-time fluorescent quantitative PCR to detect the mRNA expression of inflammatory factors and Western blot to test the changes of phosphorylated protein within the signaling pathway of NF-κB/MAPK.<p>RESULTS: Compared with normal cells, the survival rate of ARPE-19 cells was decreased after the LPS stimulation. With the increase of exogenous LBP concentration, the survival rate of ARPE-19 cell was gradually increased, while the inflammatory factors expression of cytokines IL-1β, IL-6 and MCP-1 were reduced accompany with the phosphorylated proteins(p-p65, P-IκBα, p-JNK, p-ERK and p-p38)of NF-κB/MAPK signaling pathway were decreased.<p>CONCLUSION: LBP prevents LPS-induced inflammatory response of ARPE-19 by inhibiting the intracellular inflammatory factors and the phosphorylation of the related protein within NF-κB/MAPK signaling pathway.

Protective effect of Lycium barbarum polysaccharide on reproductive system injury of female rats induced by 2,4-dichlorophenoxyacetic acid / 环境与职业医学

Background 2,4-Dichlorophenoxyacetic acid (2,4-D) is widely used as a broad-leaved herbicide and plant growth regulator. Related studies have shown that 2,4-D has neurotoxicity, ability to disrupt endocrine function, genotoxicity, carcinogenicity, and reproductive toxicity. Objective This experiment is conducted to investigate the effect of 2,4-D exposure on reproductive system of female rats, and to preliminarily explore the potential ameliorative effect of Lycium barbarum polysaccharide (LBP) and its possible mechanism. Methods Twenty-four SPF female SD rats with six rats in each group were randomly divided into a blank control group (deionized water 1 mL·d−1), an exposure group (75 mg·kg−1 2,4-D), an LBP control group (50 mg·kg−1 LBP), and an LBP intervention group (75 mg·kg−1 2,4-D + 50 mg·kg−1 LBP). The rats were given intragastric administration once a day for 28 consecutive days. Body weight was measured every two days. After exposure, ovary and uterus were weighed and organ coefficients were calculated; the pathological changes of ovary and uterus were detected by hematoxylin-eosin staining (HE); the level of estradiol (E2) in serum was detected by ELISA; the levels of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and malondialdehyde (MDA) in serum were measured by corresponding kits; the apoptosis of ovarian and uterine cells was detected by TUNEL fluorescence staining; and the protein expression levels of Fas, FasL, FADD, Pro-Caspase-8, Cleaved-Caspase-8, Pro-Caspase-3, and Cleaved-Caspase-3 in ovarian tissues were detected by Western blotting. Results Compared with the blank control group, the ovarian structure of the exposure group was abnormal, the number of follicles at different developmental stages decreased, morphological changes were observed, and the number of atretic follicles increased; the endometrium was incomplete, with different degrees of nuclear pseudostratification and decreased number of glands in lamina propria. Compared with the exposure group, the ovarian structure of the LBP intervention group was complete, and the follicles at different developmental stages increased in amount, remained intact, and were arranged closely; the uterine structure was relatively intact, showing decreased endometrial loss and nuclear pseudostratification. There were significant differences in the levels of SOD, GSH-Px, E2, and MDA among the four groups (F=86.1, 26.2, 43.3, and 22.3, all P<0.01). Compared with the blank control group, the levels of serum SOD, GSH-Px, and E2 decreased in the exposure group (P<0.01), while the concentration of MDA increased (P<0.01). Compared with the exposure group, the levels of serum SOD, GSH-Px, and E2 in the LBP intervention group increased (P<0.01), and the concentration of MDA decreased (P<0.01). There were significant differences in the apoptosis rates of ovarian and uterine cells among the four groups (F=64.8, 55.5, both P<0.01). Compared with the blank control group, the apoptosis rates of ovarian and uterine cells increased in the exposure group (P<0.01). Compared with the exposure group, the apoptosis rates of ovarian and uterine cells decreased in the LBP intervention group (P<0.01). There were significant differences in the expression levels of death receptor pathway-related proteins in ovarian tissues among the four groups (all P<0.05). Compared with the blank control group, the expression levels of Fas, FasL, FADD, Pro-Caspase-8, Cleaved-Caspase-8, Pro-caspase-3, and Cleaved-Caspase-3 were increased in the exposure group (P<0.05 or 0.01). Compared with the exposure group, the expression levels of above proteins were decreased in the LBP intervention group (P<0.05 or 0.01). Conclusion The study findings reveal that 2,4-D can induce oxidative stress and further mediate Fas-FasL pathway to induce apoptosis, resulting in reproductive system damage in female rats. LBP can reduce the oxidative stress level, down-regulate the expression of Fas-FasL pathway-related proteins, and reduce the apoptosis of germ cells, therefore protecting reproductive system of female rats.

Lycium barbarum polysaccharide inhibits the proliferation and promots the apoptosis of osteosarcoma HOS cells by blocking Shh signaling pathway / 中国临床药理学与治疗学

AIM: To study the effects and mechanism of Lycium barbarum polysaccharide on the proliferation and apoptosis of osteosarcoma HOS cells. METHODS: Osteosarcoma HOS cells were divided into four groups: control group, LBP-I group, LBP-II group and LBP-III group. MTT was used to detect cell proliferation; PI single staining was used to detect cell cycle; Annexin V-FITC/PI double staining was used to detect apoptosis; Western blot was used to detect the expression of Cleaved Caspase-3, Cleaved PARP, cyclin-dependent kinase 4 (CDK4), cyclin D1, Shh and Gli1. Shh signal activator and Lycium barbarum polysaccharide treated osteosarcoma HOS cells together; the changes of cell proliferation, cell cycle and apoptosis were observed.RESULTS: Compared with the control group, the proliferation ability of LBP-I group, LBP-II group and LBP-III group decreased, the proportion of cells in G0/G1 phase increased[(51.2±4.1)% vs. (59.1±3.2)%, (66.8±2.0)%, (72.3±3.2)%, F=72.76, P<0.001], the level of apoptosis increased[(3.9±0.3)% vs. (13.2±1.2)%, (17.6±1.3)%, (24.8±2.1)%, F=364.50, P<0.001], the expression levels of Cleaved Caspase-3, Cleaved PARP protein increased, the expression levels of CDK4, cyclin D1, Shh and Gli1 decreased (P<0.05). Compared with cells not treated with Shh signal activator, the cells treated with Shh signal activator could reverse the effect of Lycium barbarum polysaccharide on the proliferation, cycle arrest and apoptosis of osteosarcoma HOS cells. CONCLUSION: Lycium barbarum polysaccharides blocked the cell cycle of osteosarcoma HOS cells, inhibited cell proliferation and promoted cell apoptosis by inhibiting Shh signaling pathway.

Lycium barbarum polysaccharide intervenes with CD151 and matrix metalloproteinase 3 in the articular cartilage in a rabbit model of knee osteoarthritis / 中国组织工程研究

BACKGROUND: Studies have shown that Lycium barbarum polysaccharide (LBP) has the functions of anti-aging, nerve protection, anti-fatigue, blood sugar control, anti-oxidation, and anti-tumor. It may have some protective effects against osteoarthritis of the knee, but have been rarely reported. CD151 and matrix metalloproteinase 3 (MMP-3) are two common cytokines for assessing knee osteoarthritis. OBJECTIVE: To observe the effect of LBP on the expression of CD151 and MMP-3 in rabbit osteoarthritis. METHODS: Sixty-four healthy 6-month-old white rabbits were randomly divided into four groups: blank group, model group, LBP group and normal saline group. Animal models of knee osteoarthritis were made using Hulth method in the rabbits except those in the blank group. The rats in the LBP and normal saline groups were fed with normal dose of LBP and normal saline for 4 weeks, and then the articular cartilage tissues were taken from the affected side at 12 weeks after modeling. The morphological changes of the articular cartilage were observed by hematoxylin-eosin staining. The expression levels and spatial distribution of CD151 and MMP-3 in articular cartilage was observed by immunohistochemical staining and western blot. Ethic approval was given by the People’s Hospital of Ningxia Hui Autonomous Region (approval No. 2014-30817). RESULTS AND CONCLUSION: immunohistochemistry staining and western blot results showed that the absorbance values and protein expression of MMP-3 and CD-151 were significantly lower in the LBP group than the normal saline and model groups (P < 0.05). Therefore, the expression of CD151 and MMP-3 in the articular cartilage of osteoarthritis was increased, and LBP could inhibit the expression of CD151 and MMP-3 in osteoarthritis, so as to slow down the occurrence of osteoarthritis.

Effect of Lycium barbarum polysaccharides on inflammatory cytokines in type 2 diabetes mellitus model mice without myeloid differentiation factor 88 gene / 上海交通大学学报（医学版）

Objective: To investigate the effect of Lycium barbarum polysaccharides on inflammatory cytokines in type 2 diabetes mellitus (T2DM) mice without myeloid differentiation factor 88 gene (MyD88-/-). Methods: Levels of interleukin 1β (IL-1β), IL-6, IL-8, transforming growth factor β1 (TGF-β1), and IL-10 in serum were assessed by ELISA in the MyD88-/- T2DM mice which had been administered with different doses of LBP (20, 40, and 80 mg/kg). Mouse macrophages Raw264.7 were stimulated by lipopolysaccharide (LPS) after treatment with different concentrations of LBP (25, 50, and 100 μg/mL). Then Western blotting was used to detect nuclear translocation level of nuclear factor κB (NF-κB) and protein expressions of inhibitor of NF-κB (IκB) and p-IκB. Results: Serum levels of IL-1β and TGF-β1 in MyD88-/- T2DM mice were down-regulated by LBP (P<0.05). Cell experiment proved that nuclear migration of NF-κB was dose-dependently inhibited by LBP, and the level of p-IκB was reduced by high dose of LBP. Conclusion: LBP can reduce some proinflammatory cytokines in the MyD88-/- T2DM mice, which may be related with its inhibitive effect on the phosphorylation of IκB and nuclear migration of NF-κB in the macrophages.

Lycium barbarum polysaccharide protects against LPS-induced ARDS via an Akt/eNOS signaling pathway / 中国药理学通报

Aim To investigate the effects of Lycium barbarum polysaccharide ( LBP ) on LPS-induced ARDS in mice and the potential mechanisms. Methods Thirty-two C57BL/6 male mice were randomly divided into control group, LPS group, LBP group and LY294002 ( Akt inhibitor) group, with eight mice in each group. The pathological changes in lung tissues were evaluated by HE staining, pulmonary edema was measured by wet/dry ratio( W/D) , and the concentra-tions of total protein and the levels of inflammatory cy-tokines in BALF were determined. MDA and SOD lev-els in lung tissues and the apoptosis of lung tissues were detected. The protein expression levels of cleaved caspase-3, p-Akt and p-eNOS were determined by Western blot. Results Compared with control group, severe pathological lung injury changes were observed in LPS group, and the W/D ratio, levels of total pro-tein and inflammatory cytokines in BALF, levels of MDA in lung tissues and the expression of cleaved caspase-3 significantly increased(P<0.05), while the lung SOD activity, the p-Akt and p-eNOS expression decreased( P<0.05) . LBP could significantly attenu-ate the indexes above(P<0.05). However, the pro-tective effects of LBP on ARDS were inhibited by Akt inhibitor LY294002. Conclusions LBP protects a-gainst LPS-induced ARDS in mice by alleviating EC barrier dysfunction via the suppression of inflamma-tion, oxidative stress and apoptosis, at least partially via activation of the Akt/eNOS singaling pathway.

Effect of Lycium barbarum polysaccharide on atherosclerosis in mice by up-regulating adiponectin expression / 药物评价研究

Objective To explore the effect of Lycium barbarum polysaccharide (LBP) on adiponectin (APN) expression and the mechanism of lowering blood-lipid and anti-inflammation in atherosclerotic (AS) mice.Methods C57BL/6J mice with normal feed were chosen as control group.Thirty-two ApoE-/-mice with high cholesterol diet were successfully established as AS models,and then the mice were randomly divided into model group and three LBP groups,which were feed with high,medium,and low dose of LBP.After feeding for four weeks,aortic blood and tissues were collected.Blood-lipid,inflammatory factors,endothelin-1 (ET-1),APN,AdipoR1,and AMPK pathway related protein expression were detected.Differentiated 3T3-L1 cells were divided into control group,LBP group,and LBP + BML-275 group.Triglyceride (TG),inflammatory factors,APN,AdipoR1,and AMPK pathway related protein expression was investigated.Results In mice,compared with the control group,typical AS pathomorphologic changes were found in aorta and the levels of TG,total cholesterol (TC),nitric oxide (NO),ET-1,interleukin-6 (IL-6),and tumor necrosis factor (TNF-α) in the model group were significantly increased,while the protein expression of HDL-C,APN,AdipoR1,PPARα,AMPKα,and p-AMPK-α and Acyl-CoA oxidase (ACO) mRNA expression was reduced.Compared with the model group,AS pathomorphologic state was obviously improved in aorta and the amount of TG,TC,NO,ET-1,IL-6,and TNF-α in LBP groups were markedly decreased,while the protein expression of HDL-C,APN,AdipoR1,PPARα,AMPKα,p-AMPKα,and ACO mRNA expression was up-regulated.These changes were all in a dose-dependent manner.In differentiated 3T3-L1 fat cells,compared with control group,LBP enhanced the expression of APN,AdipoR1,PPARα,AMPKα,p-AMPKα,and ACO,but decreased the amount of TG,IL-6,and TNF-α.Compared with LBP group,the levels of TG,IL-6 and TNF-α was notably increased in BML-275 group.Conclusion LBP up-regulates the expression of APN and AdipoR1,activates APN/AMPK pathway,plays a role in lowering blood-lipid and anti-inflammation,and thus relieves AS in mice.

Effects of lycium barbarum polysaccharide on UV-mediated DNA strand breakage damage in HSF cells / 中华医学美学美容杂志

Objective To investigate the protective effect of lycium barbarum polysaccharide (LBP) on DNA damage of HSF cells induced by UV.Methods We established the model of UV induced photo damage in HSF cells.We detected the viability of HSF cells by using MTT colorimetry.The UV absorption spectrum of LBP was also measured by UV spectrophotometer.The level of ROS was detected by DCFH-DA fluorescent probe method.Comet assay was employed to evaluate the DNA strand breakage damage.Results When the concentration of LBP was less than or equal to 300μg/ml,there was no significant effect on the proliferation of HSF cells (P＞0.05).When the concentration was more than 300 μg/ml,it could inhibit the cell proliferative activities (P＜0.05).Compared to the UV groups,UV+LBP groups can respectively improve the cell proliferation activity (P＜0.05).The absorbance was slight range 280 from 400 nm.Compared with the UV group,the relative fluorescence intensity and the migration distance of UV+ LBP groups were significantly decreased (P＜0.05).Conclusions Lycium barbarum polysaccharide can effectively inhibit the proliferation activity and protect the breakage of DNA strand induced by UV,which is probably due to its action of removing free radicals.

Inhibitory effect of lycium barbarum polysaccharide on bleomycin-induced pulmonary fibrosis in mice and its mechanisms / 医学研究生学报

Objective Pulmonary fibrosis ( PF) is a group of pulmonary interstitial pathological changes caused by various factors, and its pathogenesis is not yet fully elucidated .This article discussed the effect of lycium barbarum polysaccharide ( LBP) on ble-omycin-induced PF in mice and its action mechanisms . Methods The C57/BL6 mice were randomly divided into 6 groups:sham-opera-tion, PF model, dexamethasone ( DM), high-dose LBP, medium-dose LBP, and low-dose LBP.The mice in the sham-operation group were injected into the trachea with isotonic saline and those in the oth-er groups with 5 bleomycin at mg/kg for establishing the PF model . Two days after modeling , the mice in the DM and high-, medium-,and low-dose LBP groups were treated with DM at 5 mg/kg and LBP at 0.8, 0.4, and 0.2 g/kg, while those in the sham-operation and PF model groups with the same volume of isotonic saline , respectively , qd, for 4 consecutive weeks .Then, the pathological chan-ges of the lung tissue were observed and the hydroxyproline ( HYP) content in the lung tissue was detected for all the animals .RT-PCR was used to determine the relative expressions of the PF-related COL1A1 and α-SMA genes. Results Compared with the PF mod-els, the the high-, medium-, and low-dose LBP groups showed significant increased body weight after 4 weeks of medication ([14.29 ±0.38] vs [16.12 ±0.37], [15.58 ±0.25] and [15.07 ±0.21] g, P<0.01), the high-and medium-dose groups ex-hibited remarkably decreased lung indexes ([0.887 ±0.13] vs [0.847 ±0.22] and [0.859 ±0.18]%, P<0.05), and the high-dose group presented markedly reduced alveolitis score (3.40 ±0.23 vs 3.09 ±0.22, P<0.05), PF score (3.57 ±0.27 vs 3.07 ± 0.31, P<0.01) and HYP content ([0.831 ±0.05] vs [0.786 ±0.07] μg/mg wet weight, P<0.05).In comparison with the mod-el group, the DM, high-dose LBP and medium-dose LBP groups showed significantly decreased gene expressions of COL 1A1 (1.53 ± 0.13 vs 1.26 ±0.10 and 0.98 ±0.17, P<0.05) and α-SMA (5.67 ±0.47 vs 4.19 ±0.28 and 2.29 ±0.31, P<0.05). Conclusion LBP can suppress the progression of bleomycin-induced pulmonary fibrosis by inhibiting the gene expressions of COL 1A1 andα-SMA and decreasing the HYP content in the lung tissue .

Inhibitory effect of Lycium barbarum polysaccharide on tumor growth and metastasis in MMTV-PyMT mouse model of breast cancer / 中国实验动物学报

Objective To investigate the inhibitory effect of Lycium barbarum polysaccharide ( LBP) on the tumor growth and metastasis in MMTV?PyMT mouse model of breast cancer. Methods The population of MMTV?PyMT trans?genic mice was expanded and identified. 8?week old MMTV?PyMT?positive female mice were randomly divided into LBP group and control group, 8 mice in each group. The mice of LBP group were given LBP treatment (50 mg/kg, i. p. ), and the control group was given normal saline in the same volume, once every 2 days for 4 weeks. The tumor size was measured every two days. The mice were killed at 4 weeks after treatment, the lungs were removed and fixed in Bouin′s solution to observe the number of metastatic nodules, and tumor tissues were used for immunohistochemical examination of tumor cell proliferation and vascular density. Results The tumor formation rate was 100% in the MMTV?PyMT?positive mice. The tumor weight of LBP group was 4?208 ± 0?4463 g, significantly lower than the 6?477g ± 0?3724 g in the control group (P<0?005). The number of pulmonary nodules of the LBP group was 12 ± 1?155, significantly less than that of the control group (20 ± 2?745) (P<0?05). The immunohistochemical examination using Ki67 and CD31 staining showed that tumor cell proliferation and microvessel density of the LBP group were significantly less than the NS group. Conclusions LBP inhibits breast cancer growth and metastasis through the inhibitory effect on tumor growth and metastasis, inhibition of tumor cell proliferation and angiogenesis in MMTV?PyMT mice. These mice can be used as an ideal model for studies on antitu?mor drug development for the treatment of breast cancer lung metastasis.

Using quadratic regression universal rotary combination design to optimize the fluorescence labeling condition of lycium barbarum polysaccharide / 国际中医中药杂志

Objective Quadratic regression universal rotary combination design was used to optimize the fluorescence labeling condition of lycium barbarum polysaccharide (LBP).Methods The fluorescence labeling condition of LBP presented, when the covalent coupling of LBP and tyramine was reacted with fluorescein isothiocyanate (FITC). Filter the best labeling condition via using quadratic regression universal rotary combination design experiment on the relationship of labeling efficiency among pH value of buffer solution, reaction time, temperature and the dose of tyramine.Results The regression equation was:Y=0.085 41 - 0.002 82X1 - 0.015 68X2 + 0.008 11X3 + 0.005 01X4 + 0.008 75X1X2 - 0.005 75X1X3 - 0.001 75X1X4 + 0.010 63X2X3 + 0.000 125X2X4 + 0.000 25X3X4 - 0.021 44X12 - 0.008 89X22 - 0.001 984X32 + 0.003 66 X42, and the variables fromX1 toX4 represented pH value, reaction time, temperature and dose of tyramine, respectively. The goodness of fitting of regression equation was statistically significant. The condition of labling LBP was optimized when the temperature was at 50℃, timing of labling at the fourth day and pH value was 8.5.Conclusion The condition of labling LBP was optimized with suitable temperature, pH value and extended timing.

Anti-tumor effects of lycium barbarum polysaccharide on pancreatic cancer cells by polarization of macrophages / 天津医药

Objective To explore the effects of lycium barbarum polysaccharide (LBP) on restraining the mouse pancre?atic cancer cells LTPA by the polarization of macrophages to type 1 macrophages (M1). Methods LTPA tumor model of the subcutaneous CB-17SCID mice was constructed. Model mice were randomly divided into tumor-bearing model group (n=10) and LBP treatment group (n=10). The LBP treatment group was fed 10mg/kg LBP every day, and the tumor-bearing model group was fed the same dose of normal saline. The same amount of macrophages Raw264.7 was randomly divided into the control group and experimental groups (different concentrations of LBP). MTT assay was used to detect the optical density (OD) of Raw264.7 in experimental groups and control group. ELISA was used to detect the levels of the interleukin (IL)-12 and IL-10 in experimental group (LBP was 100 mg/L) and the control group. Flow cytometry was used to test the levels of the membrane protein CD16/32 and CD206 in experimental group (LBP was 100 mg/L) and the control group. The tumor mass was weighted and the volume was calculated after three weeks. The effects of LBP on the growth of subcutaneous tumor were detected. HE staining and KI-67 staining were used to detect the microscopic changes of tumor and the proliferation of the LTPA. Results The dose of 100 mg/L LBP can promote the growth of the macrophages Raw264.7 (P<0.01), and induced the high expression of CD16/32 and low expression of CD206, high secretion of IL-12 and low secretion of IL-10. The weight, volume of the tumor and the expression of KI-67 were significantly lower in experimental group than those in the con?trol group (P<0.01). The microscopic necrosis area range of tumor was larger than that of control group. Conclusion The LBP has the effect of restraining LTPA by the polarization of macrophages to M1.

Effects of lycium barbarum polysaccharides on retinal pathological change and expression of VEGF in retina of diabetic rats / 中华实验眼科杂志

Background The primary pathological basis of diabetic retinopathy (DR) is new blood formation due to anoxia and inflammation,which results in breakdown of blood-retinal barrier (BRB).Vascular endothelial growth factor (VEGF) is a key factor promoting neovascularization.Researches determined that lycium barbarum polysaccharides (LBP) can protect cells against oxidative damage.However,the study of LBP in ophthalmology is lack.Objective This study was to investigate the effects of LBP on the dynamic pathological change of retinal vessels and expression of VEGF in retina of diabetic rats.Methods One hundred and seventeen SPF male SD rats were randomly divided into the normal control group,the diabetic mellitus (DM) group and the LBP group according to random number table.Type 1 DM models were induced by intraperitoneal injection of streptozotocin (STZ,55 mg/kg) in the rats of the DM group and the LBP group,and then 250 mg/kg LBP was intragastically administered in the rats of the LBP group.The morphological change of retinal vessels was dynamically observed by retinal stretched preparation with Evans blue (EB) in 4,10 and 16 weeks after modeling.E B (45 mg/kg) was slowly injection via jugular vein,and 1％ polyoxymethylene was infused into the left ventricule.The eyeballs were extracted and retina were isolated.EB content in the retinas (mg/g) was calculated using retinal stretched preparation method at the time points mentioned above.Expressions of VEGF protein and mRNA in the retinas were detected by immunohistochemistry and real-time quantitative PCR at various time points,respectively.Results Retinal stretched preparation with EB exhibited that the abnormal degree in the shape,diameter of vessels and leakage of the retinal blood vessels were significantly slighter in the LBP group than those of the DM group in 4,10,16 weeks after modeling.At 4,10,16 weeks,EB content in the retinas was (12.17±1.55),(16.46±1.60) and (19.55±1.49) mg/g,which was significantly lower than (15.76± 1.90),(21.61 ±2.05) and (26.30±2.28) mg/g of the DM group (P＜0.05).Immunochemistry showed that the expression of VEGF protein primarily located at retinal ganglion cells (RGCs) layer.The staining intensity for VEGF protein was weaker in the LBP group than that of the DM group.The expression levels of VEGF protein (A value) in the LBP group were 0.234±0.011,0.331±0.023 and 0.536±0.031at various time points,with significant decline in comparison with 0.281±0.018,0.533±0.055 and 0.765±0.075 of the DM group (all at P＜0.05).Real-time quantitative PCR revealed that the expression levels of VEGF mRNA were 0.157±0.013,0.505 ±0.114 and 1.577±0.074 in the LBP group at various time points,which were significantly lower than 0.235±0.209,1.043±0.084 and 2.446±0.061 of the DM group (all at P＜0.05).Conclusions LBP can alleviate the DM-induced retinal vasculopathy,lessen the leakage of vessels well,and further protect the BRB.

The anti-aging effect of lycium barbarum polysaccharide on human retinal pigment epithelial cell / 中华实验眼科杂志

Background The pathogenesiof age-related maculadegeneration (AMD) iassociated with the senility of human retinal pigmenepithelium (RPE) cells.Seeking drug to arresRPE cell senility iof significance fothe prevention and treatmenof AMD.Research showed thathe lycium barbarum polysaccharide (LBP) can delay senility,buitinfluence on RPE cell aging iunclear.Objective Thistudy wato discusthe protective effecand mechanism of LBP on RPE cell aging.MethodPorcine retinal neural epithelial layewaisolated,and photoreceptooutesegmen(POS) waextracted by density gradiencentrifugation and marked by FITC.The POwathen co-cultured with RPE cellin the medium containing 0.01,0.10 and 1.00 g/L LBP fo24 hours.The areof fluorescence,representing the amounof POphagocytosed by RPE cells,wameasured undethe fluorescenmicroscope to evaluate the influence of LBP on the phagocytifunction of RPE cells.The POS-induced RPE lipofuscin-uptake cell model waestablished by co-culturing human RPE cellwith porcine POfo3 weeks.The RPE-POco-culture cell model waincubated in medium containing 0.01,0.10 o1.00 g/L LBP,and the autofluorescence caused by lipofuscin up-taken into RPE cellwadetected with flow cytometry.cell counting kiwaused to assescell proliferation and viability (value) 24,48 and 72 hourafteculturing.ResultPorcine POpresented athin rodundethe lighmicroscope and appeared abilayedisc-like structureundethe transmission electron microscope,and itFITC-labeled yellow-green autofluorescence waobserved undethe fluorescenmicroscope.No POwaup-taken into the RPE cellin the normal control group,buthe areof POphagocytosed by RPE cellwagradually enlarged with increasing doseof LBP,showing significandifference among the group(F =21.425,P =0.006).Compared with the POcontrol group,the phagocytosed areincreased avariouconcentrationof LBP+POgroup(P＜0.01).Flow cytometry showed thathe autofluorescence value in the POcontrol group wamore highethan thaof the normal control group.Athe LBP dose increased,the autofluorescence value in the RPE celldeclined gradually and iwaneathe normal value in the 1.00 g/L LBP+ POgroup.The rate of proliferation of the lipofuscin RPE cellvaried with the increase of doseof LBP with the maximal value in the normal RPE group and minimal value in the lipofuscin RPE group,and the rate of proliferation of the lipofuscin RPE cellascended with increasing doseof LBP until neathe normal value in the 1.00 g/L LBP + lipofuscin RPE cellgroup (P＞0.05).ConclusionLBP enhance the anti-aging effecof human RPE cellby strengthening the phagocytiability to POand the ability to remove lipofuscin and by heightening the proliferation of human RPE cells.

Protective effects of Lycium barbarum polysaccharide on rat hippocampal neurons injured by oxygen-glucose deprivation and reperfusion / 中国药学杂志

OBJECTIVE: To investigate the protective effects of Lycium barbarum polysaccharide(LBP) against injury from oxygen-glucose deprivation/reperfusion (OGD/RP) in primary cultured rat hippocampal neurons. METHODS: Cultured hippocampal neurons were exposed to oxygen-glucose deprivation (OGD) for 2 h followed by a 24 h reperfusion. The MTT assay and the lactate dehydrogenase (LDH)release were used to evaluate the protective effects of LBP. Superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and malondialdehyde (MDA)were determined by spectrophotometry using commercial kits. Mitochondrial membrane potential (MMP) and the intracellular free calcium concentration ([Ca2+]i) in hippocampal neurons were measured using confocal laser scanning microscope (CLSM). RESULTS: Treatment with LBP (10-40 mg · L-1) significantly attenuated neuronal damage and inhibited LDH release in a dose-dependent manner. Furthermore, LBP enhanced activities of SOD and GSH-Px but it decreased their MDA content, inhibited [Ca2+]i elevation and decrease of MMP in ischemia-reperfusion treated hippocampal neurons. CONCLUSION: These findings suggested that LBP may be a potential neuroprotective agent for cerebral ischemia-reperfusion injury.