Research progress in immune regulation mechanism of Polygonati Rhizoma polysaccharide / 中国药理学与毒理学杂志

Polygonati Rhizoma is the dry rhizome of Liliaceae plants Polygonatum kingianum coil ethemsl, Polygona?tum sibiricum Redoute and Polygonatum cyrtonem Hua. It tastes sweet and has a flat nature. It belongs to the spleen, lung and kidney channels. Polygonati Rhizoma contains a variety of chemical components, including polysaccharides, alkaloids, steroidal saponins, lignans, phytosterols, and so on. Polygonati Rhizoma polysaccharide (PSP) is one of the main bioactive components of Polygonati Rhizoma. It is widely used. It has the effects of enhancing immunity, anti-inflammatory, anti-virus and regulating blood lipid. In recent years, the immunomodulatory function of PSP has been paid more and more attention by researchers. PSP can play an immunomodulatory role through a variety of mecha?nisms. (1) Effects of PSP on innate immunity. ① Macrophages have a strong ability to phagocytize and clear foreign bodies. When polysaccharides bind to macrophage specific membrane receptors, the immune response will be officially activated. RAW264.7 cells can be activated by PSP MR and TLR4 mediated signal pathway to improve the pinocytosis and phagocytosis of RAW264.7 cells. ② Natural killer cell (NK cell) is a very important immune cell in the body. It is a non-specific immune killer cell naturally existing in the body. It has the dual functions of immune regulation and cytotoxic?ity. It was found that the signal pathway mediated by PSP CR3 and TRL2 may play a major role in the stimulation of NK cells. (2) Effects of PSP on adaptive immune response. ① Lymphocytes can be divided into two forms: T cells and B cells due to different differentiation and maturation sites. T lymphocytes are the general name of thymus dependent lym?phocytes. B lymphocytes differentiate and mature from animal bone marrow cells and exert their humoral immune func?tion by secreting different antibodies. It was found that PSP could activate T/B lymphocytes and increase the ratio of CD4+/CD8+in lymph cells to promote the regulation of immune system.②Thymus and spleen index refers to the level of body immunity through the development of immune organs and the functional status of immune cells. The higher the index of thymus and spleen, the higher the immune activity. A large number of studies have found that PSP can improve immune activity by promoting the proliferation of spleen lymphocytes and regulating organ index, so as to increase the weight and index of thymus and spleen induced by CY. ③ Antibody is a glycoprotein secreted by B cells after antigen stimulation and a series of proliferation and differentiation into plasma cells. Antibody production level is one of the main indicators of nonspecific immune function. PSP can not only improve the serum antibody level of mice by regulating the phagocytosis of mouse macrophages and the level of serum hemolysin, but also enhance the concentration of IL-2 secreted by spleen lymphocytes in vitro to increase the level of antibody response, and then improve the humoral immune function of the body. (3) Effect of PSP on cytokines. ① A large number of experiments have proved that PSP has a significant effect on promoting the production of interleukin (IL). PSP can combine with specific receptors on the surface of immune cells to activate various intracellular signal transduction pathways, enhance the secretion of cytokines such as IL-2, IL-4, IL-6 and IL-10 by spleen lymphocytes in vitro, make them directly kill target cells and regulate the immune function of the body at the molecular level. ② Interferon (IFN) is a special protein or glycoprotein produced by human or animal cells in response to various stimuli. It plays an important role in anti-virus, immune regulation and cell proliferation control. It was found that PSP could increase IFN-γsecreted by T cells and NK cells, activate macrophages to regulate immune function. ③ Tumor necrosis factor (TNF) is mainly produced by activated macrophages, NK cells and activated T cells. It is a cytokine with important biological activity in antitumor immune response.④ Tumor necrosis factor (TNF) is mainly produced by activated macrophages, NK cells and activated T cells. It is a cytokine with important biological activity in antitumor immune response. PSP can promote the proliferation and phagocytic activity of macro?phage RAW264.7 to reduce its apoptosis rate. By increasing the secretion of TNF-α, PSP can promote the dissociation between NF-κВprotein and IκВp65 protein after phosphorylation, so as to start the expression and transcription of related immune genes. In conclusion, PSP can improve immunity and has a good application prospect in the development of immunomodulatory drugs.

Improvement Effects of Polygonatum sibiricum Polysaccharides on Myocardial Injury of Acute Myocardial Infarction Model Rats / 中国药房

OBJECTIVE：To study the imp rovement effects of Polygonatum si biricum polysaccharides（PSP）on the myocardial injury of acute myocardial infarction （AMI）model rats. METHODS ：The rats were randomly divided into blank control group ， model group ，aspirin group （positive control ，25 mg/kg），PSP low-dose ，medium-dose and high-dose groups （0.5，1，2 g/kg）， with 10 rats in each group. Except for blank control group ，AMI model was established by ligation of the anterior descending branch of the left coronary artery of rats in other groups. After modeling ，blank control group and model group were given normal saline intragastrically ，and administration groups were given relevant medicine intragastrically ，once a day ，for consecutive 28 days. After last medication ，left ventricular ejection fraction （LVEF）and left ventricular short axis shorting rate （LVFS）of rats were detected. The morphological changes of myocardial tissue were observed. The levels of oxidative stress indexes （SOD，MDA， ROS）in myocardial tissue of rats were detected by ELISA. The expression of apoptosis-related proteins （Bcl-2，Bax，caspase-3， caspase-8，caspase-9）and the Wnt/ β-catenin pathway-related proteins （Wnt1，β-catenin）in left ventricular anterior wall tissue of rats were detected by Western blot assay. RESULTS ：Compared with model group ，the levels of LVEF and LVFS ，the levels of SOD in myocardial tissue and protein expression of Bcl- 2 in left ventricular anterior wall tissue were increased significantly in PSP medium-dose，high-dose groups and aspirin group （P＜0.05）；the levels of MDA and ROS in myocardial tissue ，the protein expression of Bax ，caspase-3，caspase-8，caspase-9，Wnt1 and β-catenin in left ventricular anterior wall tissue were decreased significantly（P＜0.05）；myocardial tissue structure disorder and inflammatory cell infiltration were reduced. CONCLUSIONS ： PSP can relieve myocardial injury in AMI model rats ；its mechanism may be related to increasing SOD level in myocardial tissue ，decreasing MDA and ROS level ，regulating apoptosis-related proteins and Wnt/ β-catenin pathway-related proteins.

A new benzofuran lignan from rhizomes of Polygonatum sibiricum / 中草药

Objective: To isolate and identify the chemical constituents from the rhizomes of Polygonatum sibiricum. Methods: The compounds were isolated and purified by various modern chromatographies, and their structures were identified by physiochemical properties and spectroscopic data. Results: Five compounds were isolated from the EtOH extract of the rhizomes of P. sibiricum, which were elucidated as polygonneolignanoside A (1), (+)-isolariciresinol-9'-O-β-D-glucopyranoside (2), trans-N-p- coumaroyltramine (3), 3-(4-hydroxyphenyl)-N-[2-(4-hydroxyphenyl)-2-methoxyethyl]acrylamide (4), 3-(4-hydroxy-3-methoxyphenyl)- N-[2-(4-hydroxyphenyl)-2-methoxyethyl]acrylamide (5). Conclusion: Among them, compound 1 was a new benzofuran lignan, and compounds 2-5 were isolated from this plant for the first time.

Study on the Effects of Different Primary Processing Methods on the Quality of Polygonatum sibiricum / 中国药房

OBJECTIVE： To explore the effects of different primary processing methods on quality change of Polygonatum sibiricum， and to optimize primary processing method of P. sibiricum with different grades and shapes. METHODS： After processed with different drying methods， boiling method and steaming method， sulfuric acid-phenol method was used to determine the contents of indicator component P. sibiricum polysaccharide in samples. The contents of ethanol-soluble extract， water-soluble extract and total ash were investigated. The optimal primary processing methods were screened. RESULTS： After processed with different methods， the contents of polysaccharide （≥7％）， ethanol-soluble extract （≥45％） and total ash （≤4％） of samples at all levels conformed to the requirements of the 2015 edition of Chinese Pharmacopoeia （part Ⅰ）； from aspect of drying method， microwave drying method was better than oven drying method；for P. sibiricum with different morphologies （individual and lamellar）， the quality of lamellar samples of P. sibiricum was better than that of individual P. sibiricum. As primary processing method， steaming was better than boiling. The contents of P. sibiricum polysaccharide were in relatively high level， when the steaming and boiling time of gradeless and uniformly-priced goods of P. sibiricum were 20 and 15 minutes respectively. CONCLUSIONS： By discussing the different primary processing methods， high-efficient primary processing method for the individual and lamellar samples of P. sibiricum as well as gradeless and uniformly-priced goods of P. sibiricum are preliminarily obtained， which can provide a scientific basis for the effective utilization of P. sibiricum resources.

Colonization investigation of arbuscular mycorrhizal fungi (AMF) and dark septate endophytes (DSE) in roots of Polygonatum kingianum and their correlations with content of main functional components in rhizomes / 中草药

Objective: To investigate the colonization of arbuscular mycorrhizal fungi (AMF) and dark septate endophytes (DSE) in the roots of Polygonatum kingianum planted at different areas, and explore the correlations between the colonization rate and the main functional components. Methods: Fresh roots of P. kingianum from five plots in Yunnan Province were taken as research objects. The alkali dissociation method was used to investigate the colonization of AMF and DSE in the roots of P. kingianum. Then the morphological structure was photographed. The content of P. sibiricum polysaccharides, diosgenin, total flavonoids and extractum were separately determined by phonel-sulfate method, colorimetry method of vanillin-acetic acid-perchloric acid, spectrophotometry method with rutin standard and hot dipping method with alcohol. The correlations between the colonization rates of AMF or DSE and four main functional components were analyzed by Pearson correlation coefficient method. Results: The average colonization of AMF and DSE in the five plots were 26.25%-57.54% and 31.67%-45.19%, respectively. The colonization rates of AMF and DSE from HHMZ was higher than the other four others. All of correlations among the colonization rates of AMF, DSE and their typical structure and the four main functional components in the rhizomes were positive correlation, in which the correlations between the content of polysaccharides, diosgenin or total flavonoids and the colonization rates of AMF or DSE were higher, and their correlation coefficient were respectively 0.838/0.887, 0.819/0.703, and 0.785/0.855 (AMF/DSE). Furthermore, the correlations between the content of polysaccharides and the colonization rates of AMF hypha or DSE were significant. In addition, there were high correlations among the colonization rates of AMF, DSE and their typical structures, as well as the content in pairs of four functional components. Conclusion: There were higher colonization rates of AMF and DSE in the fibrous roots of P. kingianum. The correlations between the colonization rates of AMF or DSE and each main functional component were positive. This study provides data support and experimental basis for the implementation of ecological planting of P. kingianum and the use of biological means to increase production and income.rates

Study on content of polysaccharide during processing of Polygonatum sibiricum by RSM and PMP-HPLC characteristic map technology / 中草药

Objective: To investigate the best extracting conditions of Polygonatum sibiricum polysaccharides by single and compound enzymatic methods, and to study on the change content of polysaccharide in processing of P. sibiricum by steamed and shined for nine times, which provide theoretical basis for processing of P. sibiricum and other Chinese herbal medicines. Methods: Response surface method (RSM) was used to optimize the extraction of polysaccharides in P. sibiricum. PMP precolumn derivatization and HPLC characteristic map technology were used to identify and analyze the changes of polysaccharide of P. sibiricum being steamed and shined for nine times and establish polysaccharide characteristic map of P. sibiricum by ancient processing and distillate. The effect of different enzymes on the extraction efficacy of P. sibiricum polysaccharides was also compared. Results: The extracting effect of cellulase in single enzyme was the best. Moreover, cellulose and pectinase were also the most significant factor for extracting polysaccharides from P. sibiricum in four complex enzymes. The best extracting conditions for polysaccharide obtaining by RSM as follows: Cellulase temperature was 51 ℃, cellulase dosage was 2%, and the total polysaccharide content was (675.34 ± 0.10) mg/g when the pH value of enzymolysis was 5.40. PMP-HPLC was used to detect four processed P. sibiricum, which contained mannose, ribose, rhamnose, glucose, and galactose. Glucose had a high proportion in P. odoratum, P. cyrtonema, and P. sibiricum polysaccharide, and P. filipes polysaccharide mainly composed of mannose. With the increase of processed times, the content of each monosaccharide increased first, then decreased, then increased and then tended to be stable. The content of mannose and galactose varied greatly. The content of monosaccharides in the distillate increased first and then stabilized, and the content changes of mannose and glucose were obvious. Conclusion: The extraction efficiency and purity of P. sibiricum polysaccharides can be improved by single and compound enzymatic extraction with cellulase, and the extraction yield of polysaccharide in P. sibiricum could be affected by steamed and shined for nine times. PMP-HPLC characteristic map technology can be used to determine the content of the polysaccharide in processing of P. sibiricum accurately and reproducibly, which can be used for the quality evaluation of P. sibiricum and other Chinese herbal medicines.

Effect of polysaccharides from Polygonatum sibiricum on lipid-metabolism related mRNA and protein expression in hyperlipidemic mice / 中国中药杂志

To study the effect of polysaccharides from Polygonatum sibiricum on mRNA and protein expressions of blood lipid metabolism in hyperlipidemic mice. The mice were randomly divided into 6 groups, namely the blank control group, the hyperlipidemia model group, the simvastatin group, and low, middle and high-dose PSP groups (200, 400, 800 mg·kg⁻¹·d⁻¹). Each group of the mice was administrated intragastrically for 14 days, respectively. Subsequently, every group of mice, except for the blank control group, was intraperitoneally injected with 75% fresh egg yolk emulsion for establishing the hyperlipidemic mice model. Upon completion of the administration, the contents of TC, TG, LDL-C and HDL-C in serum of each group were investigated in details. In particular, the mRNA expression levels of PPAR-α, PPAR-, PPAR-, SREBP-1c, IL-6 and TNF-α of the liver tissues were detected by Real-time PCR, and the protein expression levels (including PPAR-α, PPAR-, PPAR-, SREBP-1c, IL-6, TNF-α) were examined by Western blot. Consequently, the obtained results showed that the contents of the serum TC, TG, LDL-C of low, middle and high-dose PSP groups significantly decreased compared with those of the hyperlipidemia model group. Simultaneously, there were significant differences between middle-dose and high-dose PSP groups (<0.01). In striking contrast, the contents of serum HDL-C of low, middle and high-dose PSP groups significantly increased, while obvious differences were also observed between middle-dose and high-dose PSP groups (<0.01). Moreover, middle-dose and high-dose PSR groups could up-regulate the protein and mRNA expressions of PPAR-α, PPAR- (<0.05) compared with those of the hyperlipidemia model group, and down-regulate the expressions of PPAR-,SREBP-1c, IL-6 and TNF-α(<0.05) compared with those of liver tissues of the hyperlipidemia model group. In conclusion, all of the above results suggested that PSP could inhibit the oxidation of the liver lipid, and regulate the expression levels of the corresponding genes and proteins relating to the lipid metabolism, so as to play a critical role for preventing hyperlipidemia.

Inhibitory effect of Polygonatum sibiricum polysaccharides on release of inflammatory cytokines of anoxia/reoxygenation H9c2 myocardial cells through TLR4-MyD88-NF-κB signaling pathway / 中国药理学通报

Aim To assess the regulatory effects of Po-lygonatum sibiricum polysaccharides(PSP)on Toll-like receptor 4 (TLR4 )-myeloid differentiation factor 88 (MyD88)-nuclear factor κB(NF-κB)signaling path-way in anoxia /reoxygenation-H9c2 myocardial cells. Methods The H9c2 myocardial cells cultured in vitro were randomly divided into groups:control group (C group),hypoxia /reoxygenation group (H/R group), PSP group,TLR4 inhibitor group(TAK-242 group)and PSP +TLR4 inhibitor group(PSP +TAK-242 group). The cells were cultured in normal condition for 27 h in C group.The cells were subjected to 21 h hypoxia fol-lowed by 6 h reoxygenation in H/R.Definitely,the cells in TAK-242,PSP and PSP +TAK-242 groups were treated with PSP and TAK-242 with the final con-centration of 1 .5 g·L -1 and 1 μmol·L -1 for 1 2 h before 21 h hypoxia,then the cells were exposed to the normal culture condition for another 6 h.After the treatment,cell survival rate was tested by MTT method. The contents of tumor necrosis factor-α(TNF-α)and interleukin-1 β(IL-1 β)were determined by enzyme-linked immunosorbent assay(ELISA).The protein ex-pression levels of NF-κB and inhibitor κBα(IκBα) were detected by Western blot,and the expression lev-els of TLR4 and MyD88 mRNA were detected by fluo-rescence quantitative PCR method.Results Compared with H/R group,the cell survival rates were significant-ly increased,while the inflammatory cytokines contents and NF-κB protein expression were dramatically de-creased in groups PSP,TAK-242 and PSP +TAK-242, whereas the NF-κB expression was significantly down-regulated,and the IκBα protein expression was in-creased.The mRNA expression levels of TLR4 and MyD88 were markedly decreased.Conclusion PSP might protect H9c2 myocardial cells against H/R inju-ry,which may be associated with the inhibition of TLR4-MyD88-NF-κB pathway.

Advances in studies on chemical constituents of three medicinal plants from Polygonatum Mill. and their pharmacological activities / 中草药

Polygonti Rhizome belongs to the family Liliaceae. Its tuber has the medicinal functions of tonifying the kidney to arrest spontaneous emission (Bushen Yijing) and replenishing yin essence and moisturizing the viscera (Ziyin Runzao), and has been used for many years in Chinese medicine. This paper mainly summarizes the research advances on chemical constituents and pharmaco- logical activities of P. sibiricum, P. kingianum, and P. cyrtonema accepted in Chinese Pharmacopoeia, and provides the reference for their further development and utilization.

Effect of Polygonatum sibiricum polysaccharides on inflammatory raction mediated by NF-κB and myocardium tissue morphology in acute myocardial infarction rats / 中草药

Objective: To observe the effect of Polygonatum sibiricum polysaccharides (PSP) on the inflammatory raction mediated by NF-κB and myocardium tissue morphology in acute myocardial infarction (AMI) rats, and to explore the myocardial protective effect and its mechanisms. Methods: The AMI model was established by closing the left anterior descending coronary artery of SD rats, Sham group threading without ligation, then the model rats were randomly divided into model group, high-, mid-, low-dose PSP (900, 450, and 225 mg/kg) groups, and positive group (Shexiang protect myocardial pills, 12.2 mg/kg), with 10 rats in each group, ig administered once daily for 28 d. ECG S-T changes were measured before surgery, immediately after surgery, and 1, 3, 7, 10, 14, 21, 28 d after surgery in each group; The levels of inflammatory cytokines IL-6, TNF-α, NF-κB, CKMB, and LDH in serum were measured by ELISA; The myocardial tissue morphology was measured by HE staining. Results: Compared with the model group, PSP inhibited S-T segment elevation of ECG in AMI rats; The serum levels of TNF-α, IL-6, NF-κB, CKMB, and LDH in high- and mid-PSP groups decreased significantly (P < 0.05, 0.01), and the degree of myocardial injury was significantly improved. Conclusion: PSP could improve the myocardial injury, reduce the inflammation, and repair the ischemic myocardium in AMI rats. The mechanisms may be related to regulating the inflammatory raction mediated by NF-κB.

Isolation and identification of endophytic fungi from Polygonatum sibiricum in Mountain Tai and study on their antimicrobial activity / 中草药

Objective: To study the diversity of endophytic fungi from Polygonatum sibiricum growing in Mountain Tai and screen the endophytic fungi with antimicrobial activity. Methods: Endophytic fungi were isolated with the conventional method and their antimicrobial activities against tested microorganisms were investigated by using the paper disc method. The endophytic fungi with higher antimicrobial activity were identified based on ITS sequence analysis. Results: A total of 32 endophytic fungi strains were isolated from the roots, stems, leaves, and fruits of P. sibiricum belonging to seven genera. The most widely distributed genera is Fusarium sp. (31.3%), followed by Sterile mycelia (28.1%) and Alternaria sp. (21.9%). Some endophytic fungi showed a certain degree of tissue preference. More species and strains were isolated from the roots, followed by those in the stems and leaves but those in the fruits were the least. Fourteen strains from five genera with the antimicrobial activity were obtained, among which eight strains showed broad-spectrum antimicrobial activity. The strain HJG-10 with the highest antimicrobial activity was determined to be Schizophyllum commune, according to its molecular analysis. Conclusion: The research not only provides the theoretical basis for developing and utilizing the resources of endophytic fungi in P. sibiricum in Mountain Tai but also shows a new path for searching novel antimicrobial resource.