Effect of polydatin on the proliferation and apoptosis of THP-1 cells and the mechanism / 中国当代儿科杂志

OBJECTIVES@#To explore the effect of polydatin on the proliferation and apoptosis of acute monocytic leukemia cell line THP-1 and the possible mechanism.@*METHODS@#After THP-1 cells were treated with polydatin at gradient concentrations for 24 hours and 48 hours, their proliferation was determined by CCK-8 assay, and half maximal inhibitory concentration (IC50) was calculated. Logarithmically growing THP-1 cells were divided into two groups, a polydatin treatment group (treated with IC50 of polydatin) and a blank control group (treated without polydatin solution), and incubated for 48 hours. Cell apoptosis and cell cycle were measured by flow cytometry. The expression levels of PI3K, AKT, p-AKT, mTOR, p-mTOR, p70 S6K, and p-p70 S6K proteins were measured by Western blotting.@*RESULTS@#After treatment with polydatin, the proliferation of THP-1 cells was strongly inhibited, and the IC50 at 48 hours was 1 800 μmol/L. After treatment with 1 800 μmol/L polydatin solution for 48 hours, the apoptosis rate of THP-1 cells increased significantly compared with the blank control group (P<0.05). The cell cycle was arrested in the G0/G1 and S phases, with a significantly increased proportion of cells in the G0/G1 phase and a significantly decreased proportion of cells in the S phase, as compared with the blank control group (P<0.05). The expression levels of PI3K, AKT, p-AKT, mTOR, p-mTOR, p70 S6K, and p-p70 S6K proteins decreased significantly compared with the blank control group (P<0.05).@*CONCLUSIONS@#Polydatin can effectively inhibit the proliferation, block the cell cycle, and induce the apoptosis of THP-1 cells, which may be related to inhibition of the PI3K/AKT/mTOR signaling pathway.

Polydatin improves intestinal barrier injury after traumatic brain injury in rats by reducing oxidative stress and inflammatory response via activating SIRT1-mediated deacetylation of SOD2 and HMGB1 / 南方医科大学学报

OBJECTIVE@#To investigate the protective effect against intestinal mucosal injury in rats following traumatic brain injury (TBI) and explore the underlying mechanism.@*METHODS@#SD rat models of TBI were established by fluid percussion injury (FPI), and the specimens were collected at 12, 24, 48, and 72 h after TBI. Another 15 rats were randomly divided into shamoperated group (n=5), TBI with saline treatment (TBI+NS) group (n=5), and TBI with PD treatment (TBI+PD) group (treated with 30 mg/kg PD after TBI; n=5). Body weight gain and fecal water content of the rats were recorded, and after the treatments, the histopathology of the jejunum was observed, and the levels of D-lactic acid (D-LAC), diamine oxidase (DAO), ZO-1, claudin-5, and reactive oxygen species (ROS) were detected. Lipid peroxide (LPO) and superoxide dismutase (SOD) 2 content, jejunal pro-inflammatory factors (IL-6, IL-1β, and TNF- α), Sirt1 activity, SOD2 and HMGB1 acetylation level were also determined after the treatments.@*RESULTS@#The rats showed significantly decreased body weight and fecal water content and progressively increased serum levels of D-LAC and DAO after TBI (P < 0.05) with obvious jejunal injury, significantly decreased expression levels of ZO-1 and claudin-5, lowered SOD2 and Sirt1 activity (P < 0.05), increased expression levels of LPO, ROS, and pro-inflammatory cytokines, and enhanced SOD2 and HMGB1 acetylation levels (P < 0.05). Compared with TBI+NS group, the rats in TBI+PD group showed obvious body weight regain, increased fecal water content, reduced jejunal pathologies, decreased D-LAC and DAO levels (P < 0.05), increased ZO-1, claudin-5, SOD2 expression levels and Sirt1 activity, and significantly decreased ROS, LPO, pro-inflammatory cytokines, and acetylation levels of SOD2 and HMGB1 (P < 0.05).@*CONCLUSION@#PD alleviates oxidative stress and inflammatory response by activating Sirt1-mediated deacetylation of SOD2 and HMGB1 to improve intestinal mucosal injury in TBI rats.

Heterologous expression of a novel β-glucosidase BglD2 and its application in polydatin-hydrolyzing / 生物工程学报

A novel β-glucosidase BglD2 with glucose and ethanol tolerant properties was screened and cloned from the deep-sea bacterium Bacillus sp. D1. The application potential of BglD2 toward polydatin-hydrolyzing was also evaluated. BglD2 exhibited the maximal β-glucosidase activity at 45 °C and pH 6.5. BglD2 maintained approximately 50% of its origin activity after incubation at 30 °C and pH 6.5 for 20 h. BglD2 could hydrolyze a variety of substrates containing β (1→3), β (1→4), and β (1→6) bonds. The activity of β-glucosidase was enhanced to 2.0 fold and 2.3 fold by 100 mmol/L glucose and 150 mmol/L xylose, respectively. BglD2 possessed ethanol-stimulated and -tolerant properties. At 30 °C, the activity of BglD2 enhanced to 1.2 fold in the presence of 10% ethanol and even remained 60% in 25% ethanol. BglD2 could hydrolyze polydatin to produce resveratrol. At 35 °C, BglD2 hydrolyzed 86% polydatin after incubation for 2 h. Thus, BglD2 possessed glucose and ethanol tolerant properties and can be used as the potential candidate of catalyst for the production of resveratrol from polydatin.

Polydatin Ameliorated Colonic Inflammation via Precluding Phosphorylation of PKCθ/STAT3 Pathway to Inhibit Th17 Cells in Mice with Ulcerative Colitis / 中国实验方剂学杂志

Objective:To investigate the therapeutic effect of polydatin on ulcerative colitis （UC） in mice and its regulation of protein kinase C<italic>θ</italic>（PKC<italic>θ</italic>）/signal transducer and activator of transcription 3（STAT3） signaling on T helper cell 17（Th17） and its mechanism in the treatment of UC. Method:The 32 male C57BL/6 mice were randomly divided into normal group， model group， polydatin group （0.045 g·kg^-1） and sulfasalazine group （0.5 g·kg^-1）. The UC model was established by giving 3% dextran sodium sulfate （DSS） solution to free drinking water in mice. Polydatin and sulfasalazine groups were given by gavage 0.5 h before modeling for 7 days. The normal group and model group were given the same amount of normal saline. After the last administration， the colonic tissue was taken and hematoxylin-eosin （HE） was used to observe the pathological changes of colonic tissue. Flow cytometry was used to detect the proportion of Th17 in the lamina propria of colonic mucosa. The expression of interleukin-17A （IL-17A） in serum was detected by enzyme-linked immunosorbent assay （ELISA）. Polydatin was added to CD4⁺T cells purified from spleen of C57BL/6 mice by magnetic-activated cell sorting （MACS） under the stimulation of cell stimulation cocktail <italic>in vitro </italic>in order to detect its impact on PKC<italic>θ</italic> and STAT3 phosphorylation. Result:Compared with normal group， the body weight was significantly decreased， and disease activity index （DAI） scores of the model group was significantly increased （<italic>P</italic><0.01）， the colonic mucosal epithelium was damaged and inflammatory cells infiltration in the mucosa and submucosa was obvious， the proportion of Th17 in the lamina propria of colonic mucosa was significantly increased （<italic>P</italic><0.01）， and the content of serum IL-17A was significantly increased （<italic>P</italic><0.01）. Compared with the model group， the weight and DAI score of polydatin and sulfasalazine groups were significantly improved （<italic>P</italic><0.01）， the degree of colon tissue damage was significantly improved， the proportion of Th17 in colon mucosa lamina propria was significantly decreased （<italic>P</italic><0.01）， and the content of IL-17A in serum was significantly decreased （<italic>P</italic><0.01）. <italic>In vitro</italic> experiments showed that polydatin could significantly inhibit the phosphorylation of PKC<italic>θ</italic> and STAT3 in Th17 （<italic>P</italic><0.01） as well as IL-17A secretion. Conclusion:Polydatin can improve the ulcerative colitis in mice via inhibiting the phosphorylation of PKC<italic>θ</italic> and STAT3 to preclude IL-17A secreting in Th17.

Research Progress on Pharmacological Effect and Mechanism of Polydatin / 中国实验方剂学杂志

Polydatin， a polyphenolic compound， is the main active component of Chinese medicine Polygoni Cuspidati Rhizoma et Radix and has a variety of pharmacological activities. In recent years， there are more studies on the pharmacological effects and mechanisms of polydatin. Modern pharmacological studies show that polydatin has protective effects on the nervous system， cardio-cerebral vascular system， and respiratory system， and also has significant effects on the liver， kidney， lung， and other organs. Its effect of regulating blood glucose and blood lipid on atherosclerosis is significant， and the anti-fibrosis effect is significant on the liver and kidney. Polydatin can inhibit many types of tumor cells， suppress proliferation and induce apoptosis of tumor cells. Polydatin can also resist inflammation and radiation， protect bone marrow， and promote wound healing. Based on the literature on the pharmacological effects of polydatin， the authors found that the single pharmacological mechanism of polydatin is often regulated by multi-target proteins and multiple pathways， but the most of action targets are unclear， which needs to be further investigated. This study summarized the research progress on the pharmacological action and mechanism of polydatin in the past five years and put forward some suggestions on its present research situation and future research direction to broaden the research ideas of researchers and speed up the identification of the targets of its pharmacological effect. This study is expected to provide a scientific theoretical basis for the further development and utilization of polydatin.

Protective Effect of Polydatin Complicated with Emodin on Hyperuricemia Model Rats with Median Effect Principle / 中国药房

OBJECTIVE：To s tudy the protective effect of polydat in complicated with emodin on hyperuricemia （HUA）model rats，and to screen the optimal complication proportion and investigate the potential mechanism. METHODS ：SD rats were randomly divided into normal control group ，model group ，benzbromarone group （positive control ，8 mg/kg），polydatin alone group，emodin alone group and drug combination group A ，B，C，with 5 rats in each group of each dosage. Normal control group and model group were given constant volume of 0.3％ CMC-Na solution ，administration groups were given relevant medicine intragastrically. Each group was given 0.1 mL/10 g intragastrically once a day ，for consecutive 7 d. Expect for normal control group，other groups were given intraperitoneal injection of potassium oxonate 300 mg/kg 1 h before last medication to induce HUA model. One hour after last medication ，the serum contents of uric acid （UA）were determined in normal control group ，model group，benzbromarone group ，polydatin/alone group （0.625，1.25，2.5，5，10 mg/kg）and drug combination group A [the dose of polydatin+emodin were （0.625+0.625），（1.25+1.25），（2.5+ 2.5），（5 + 5），（10 + 10） mg/kg]. The effect （Fa） and combination index （CI） of above single drug groups and combination groups were calculated by the median effect principle. The dose-effect relationship curves of twocomponents alone or combination were drawn ；Fa-CI curves after simulation were als o drawn to evaluate the effect of two-drug combination. Serum contents of UA in rats were determined and Fa value was calculated in single drug groups and drug combination group B ，C [the dose of polydatin+emodin were （0.625+ 0.625），（0.625+1.25），（0.625+2.5），（0.625+5），（0.625+10）mg/kg and （0.625+0.625），（1.25+0.625），（2.5+0.625），（5+ 0.625），（10+0.625）mg/kg]. The optimal complication proportion of two drugs were screened. The serum contents of xanthine oxidase（XOD）in rats were determined in normal control group ，model group ，benzbromarone group ，polydatin/emodin alone group（10 mg/kg）and the optimal complication proportion groups. The mechanism was analyzed primarily. RESULTS ：Compared with normal control group ，the content of UA in model group was increased significantly （P＜0.01）. Compared with model group ， except for 0.625 mg/kg polydatin alone group ，the content of UA in other administration groups were decreased significantly （P＜ 0.05 or P＜0.01）. When the two drugs were used alone or in combination ，Fa value was positively correlated with drug dose （intercept＞0，correlation coefficient ＞0.9），and Fa value of the combination group was higher than that of any single drug group ； when the simulated Fa value was more than 15％，the corresponding CI value was less than 1，two-drug combination showed synergistic effect. When the complication proportion of polydatin and emodin was 1∶4，the Fa value （53.10）was similar to that of drug combination group A （53.73），and the dose of them were less [ （0.625+2.5）mg/（kg·d）vs.（2.5+2.5）mg/（kg·d）]. Compared with normal control group ，serum content of XOD in model group was increased significantly （P＜0.01）；compared with model group，serum content of XOD in administration groups were decreased significantly ，and the optimal complication proportion group was significantly lower than polydatin alone group and emodin alone group （P＜0.05 or P＜0.01）. CONCLUSIONS ：The polydatin and emodin used alone or in combination can reduce the serum content of UA in HUA model rats by inhibiting the generation of XOD. They have a certain synergistic effect ，and the optimal complication proportion is 1∶4.

Content Changes of Eight Components in Tetrastigmatis Hemsleyani Radix of Different Months / 中国实验方剂学杂志

Objective:To establish a HPLC method for determination of chlorogenic acid,rutin,polydatin,kaempferol-3-O-rutinoside,astragalin,resveratrol,quercetin,kaempferol in Tetrastigmatis Hemsleyani Radix,in order to study the content changes of eight components of different months. Method:Zorbax SB C18column (4.6 mm×250 mm,5 μm)was adopted with mobile phase consisting of acetonitrile(A)and 0.1%phosphoric acid(B)in gradient elution(0-30 min,10%-30%A;30-40 min,30%-95%A;40-45 min,95%A;45-60 min,95%-10%A). The flow rate was 0.8 mL·min-1,the column was kept at 25℃,and the detection wavelength was 320 nm. Result:Chlorogenic acid,rutin,polydatin,kaempferol-3-O-rutinoside,astragalin,resveratrol,quercetin,kaempferol showed a good linearity within the range of 13.7-549 mg·L-1(r=0.999 0),12.6-253 mg·L-1(r=0.999 1),15.8-316 mg·L-1(r=0.999 0),14.7-147 mg·L-1(r=0.999 2),8.8-88 mg·L-1(r=0.999 1),7.9-79 mg·L-1(r=0.999 5),8.6-172 mg·L-1(r=0.999 1),8.9-89 mg·L-1(r=0.999 4). There were great differences in contents of the eight flavonoid active components in different growth phases. In July and August,the relative contents of chlorogenic acid,rutin,kaempferol 3-O-rutinoside and polydatin were the highest. The highest relative content of quercetin was observed in June. The relative contents of resveratrol and kaempferol in April and May was higher than those in other mouths. The relative content of astragalin in November was the highest. Conclusion:It could provide abundant information for the production and quality control of Tetrastigmatis Hemsleyani Radix.

Spray drying technology of Shenpu Penyan Granule based on quality by design concept / 中草药

Design space approach was applied to optimize the spray drying process of Shenpu Penyan Granule (SPG) based on the concept of quality by design (QbD). The yield, moisture content, the content of paeoniflorin, chlorogenic acid, polydatin and salvianolic acid B were defined as the process critical quality attributes (CQAs). Based on the Plackett-Burman design, three critical process parameters (CPPs) including inlet temperature, specific gravity, and feeding speed were identified by using the weighted standardized partial regression coefficient method. And stepwise regression method was then used to establish the mathematical model between CQAs and CPPs in the Box-Behnken design. The variance analysis results showed that the P values of the five models were less than 0.05 and the mismatch values were all greater than 0.05, indicating that the model could describe the relationship between CQAs and CPPs. Probability based design space was obtained and verified using Monte-Carlo simulation method. According to the verification results, the robustness of first ethanol precipitation process of SPG can be guaranteed by operating within the design space parameters, which helps to improve the quality uniformity between batches of phenol extracts and provides data support for industrialization production.

Polydatin attenuates airway inflammation in asthmatic mouse model via p38 MAPK/Nrf2/HO-1 pathway / 中国药理学通报

Aim To investigate whether polydatin re-duces airway inflammation in asthmatic mouse model and explore whether this pathway is related to p38 MAPK/Nrf2/HO-1 . Methods After the establish-ment of the OVA-induced asthmatic mouse model, the animals were injected with 30 mg·kg-1 and 45 mg· kg-1 of polydatin diluted in 0. 2 mL normal saline, while the control group was replaced by normal saline. HE, PAS and Masson staining were used to observe the pathological changes of lung tissue. Diff-Quick staining was used to classify and count the number of inflamma-tory cells in BALF. ELISA was used to detect IgE ex-pressions in BALF. The content of ROS in BALF cells was detected by DHR-123 . The activities of antioxidant enzymes SOD, CAT and MDA in BALF were detected by the enzyme-linked immunosorbent assay kit. The expression of HO-1 in lung tissue was detected by im-munohistochemistry. The protein and mRNA expres-sions of Nrf2 and HO-1 in lung tissue of mice were de-tected by Western blot and RT-PCR. Results Poly-datin treatment significantly reduced inflammatory cell infiltration mucosal secretion, goblet cell proliferation and collagen deposition in the lung tissue of mice, and decreased the number of inflammatory cells and the ex-pression of total IgE and ROS in BALF. It also in-creased the levels of antioxidant enzymes such as SOD and CAT, and lowered the level of MDA. Polydatin re-duced the phosphorylation of p38 MAPK in the lung tissue of mice, enhanced the levels of mRNA and pro-tein expressions of Nrf2 and HO-1 and promoted the nuclear transfer of Nrf2 . The above effects of polydatin were dose-dependent. Conclusions Polydatin exerts anti-oxidative effects in OVA-induced asthmatic mouse model via anti-oxidant pathway. The mechanism may be achieved through the p38 MAPK/Nrf2/HO-1 path-way.

Effects of Polydatin on Protein Expression of MMP-9 and TIMP-1 in Renal Tissue of Renal Fibrosis Model Rats / 中国药房

OBJECTIVE:To investigate the effects of polydatin on protein expression of MMP-9 and TIMP-1 in renal tissue of renal fibrosis model rats,and to investigate the mechanism of delaying renal fibrosis in rats. METHODS:A total of 40 rats were randomly divided into sham operation group(normal saline),model group(normal saline),polydatin group(100 mg/kg)and benazepril group(positive control,5 mg/kg). Except for sham operation group,renal fibrosis rat model was induced by unilateral ureteral obstruction in other groups. 1 h after surgery,each group was given relevant medicine intragastrically,once a day.After 4 weeks of consecutive treatment,the contents of β 2-microglobulin(β 2-MG)and N-acetyl-β-D-glucosaminidase(NAG)in urine, serum contents of BUN and Cr were detected in each group. HE staining was used to observe and score pathological changes of renal tissue in rats. Protein expressions of MMP-9 and TIMP-1 in renal tissue of rats were determined by immunohistochemistry. RESULTS:Compared with sham operation group,the contents of β2-MG and NAG in urine,serum contents of BUN and Cr were increased significantly in model group(P<0.05),and pathological score and protein expressions of MMP-9 and TIMP-1 were also increased significantly(P<0.01). Compared with model group,protein expression of MMP-9 in renal tissue were increased continuously in polydatin group and benazepril group(P<0.01);other indexes were decreased significantly(P<0.05 or P<0.01),and there was no statistical significance in each index between polydatin group and benazepril group(P>0.05). CONCLUSIONS:Polydatin can delay renal fibrosis,the mechanism of which may be associated with up-regulating protein expression of MMP-9,down-regulating protein expression of TIMP-1 and increasing the ratio of MMP-9/TIMP-1.

Polydatin inhibits cell proliferation, invasion and migration, and induces cell apoptosis in hepatocellular carcinoma

Polydatin, a small molecule from Polygonum cuspidatum, has many biological functions, particularly anti-cancer effects. However, the anti-cancer effects of polydatin in hepatocellular carcinoma (HCC) have not been examined yet. In the present study, MTT assay, BrdU assay, transwell invasion assay, and wound healing assay were performed to determine cell proliferation, invasion and migration. Flow cytometry and TUNEL assay were used to measure cell apoptosis. Quantitative real-time PCR and western blotting assays were used to determine mRNA and protein expression levels. Xenograft experiment was performed to determine the in vivo anti-tumor effect of polydatin. Immunostaining was performed to analyze the expression of caspase-3 and Ki-67. Our results showed that polydatin inhibited cell proliferation in a concentration-dependent and time-dependent manner in the HCC cell lines. Polydatin also induced cell apoptosis in a concentration-dependent manner possibly via increasing the caspase-3 activity, and up-regulating the protein expression of caspase-3, caspase-9, Bax, and down-regulating the protein expression of Bcl-2. In addition, polydatin treatment had an inhibitory effect on cell proliferation, invasion and migration in HCC cell lines. Polydatin treatment also suppressed the Wnt/beta-catenin signaling activities in HCC cells. Polydatin treatment significantly reduced tumor growth in nude mice inoculated with HepG2 cells, suppressed the expression of Ki-67, and increased caspase-3 expression and TUNEL activity. Our data indicated the important role of polydatin for the suppression of HCC progression.

Polydatin induces human cervical cancer cell apoptosis via PI3K/AKT/mTOR signaling pathway / 中国中药杂志

To observe the effect of polydatin on proliferation and apoptosis of cervical cancer HeLa cells and explore its possible mechanism. The growth inhibitory effect was detected with MTT assay. After HeLa cells were treated with different concentrations (50, 100, 150 μmol•L⁻¹) of polydatin, MTT assay was used to detect the inhibitory effect of polydatin on proliferation of HeLa cells; Acridine orange/ethidium bromide staining was used for morphological changes in apoptotic HeLa cells； Annexin/propidium iodide staining was applied to detect HeLa cell apoptotic rate. In addition, flow cytometry was employed to analyze apoptosis and cell cycle distribution； RT-PCR and Western blot assay were used to detect PI3K, AKT, mTOR, and P70S6K mRNA and protein expression levels. The results showed that polydatin significantly inhibited HeLa cells proliferation in a dose-dependent manner. Polydatin can cause S phase arrest for HeLa cells, promote cell apoptosis and decrease the mRNA and protein expression levels of PI3K, AKT, mTOR and P70S6K. It indicated that polydatin could inhibit proliferation and induce apoptosis of cervical cancer HeLa cells, and the mechanism may be associated with inhibiting the PI3K/AKT/mTOR signaling pathway and suppressing downstream gene expression.

Effect of polydatin on inflammatory response and oxidative stress in rats with sepsis-induced acute kidney injury / 临床麻醉学杂志

Objective To determine if polydatin inhibited oxidative stress and inflammatory response in rats with sepsis-induced acute kidney injury (AKI).Methods Seventy-two rats (weighing 180-220 g) were randomly divided into the following groups: sham group, cecal ligation and puncture (CLP) CLP+normal saline group (group CN), group CLP+vehicle (group CV), and group CLP+polydatin (group CD) (n=18 each).Rats in groups CN, CV and CD underwent CLP to mimic sepsis-induced AKI.In sham group, the cecum was not ligated or punched, and the remaining procedures were the same as in group CLP.Normal saline, vehicle, and 30 mg/kg polydatin were administered at 6, 12, and 18 hours after CLP via the tail vein.At 24 hour post CLP, two clinically used markers of AKI, blood urea nitrogen (BUN), and creatinine (Cr) were tested, pathological changes of kidney tissue was observed under light microscopy in each group.Renal tubular damage assessment was carried out.Malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH) content of renal tissue, serum cytokines such as TNF-α, IL-1β and IL-6, were also measured in each group at 24 hours after CLP.Results Compared with sham group, multiple indexes such as BUN, Cr, tubular injury scores, MDA content of renal tissue, and serum cytokines incluing TNF-α, IL-1β and IL-6 increased significantly (P<0.01), while SOD and GSH levels of renal tissue significantly decreased in groups CN and CV (P<0.01).Compared with groups CN and CV, the indicators such as BUN, Cr, tubular injury scores, MDA content of renal tissue, and serum cytokines such as IL-1β and IL-6 significantly decreased (P<0.05);while SOD and GSH levels of renal tissue significantly increased (P<0.05).Conclusion Sepsis caused by sepsis cecal ligation and puncture can cause acute kidney injury.Polydatin could alleviate kidney damage by attenuating systemic inflammatory response and inhibiting oxidative stress of renal tissue.

Effect of ultrafine comminution on transdermal absorption and rheology of Polygonum cuspidatum ointment / 中草药

Objective: To study the effect of ultrafine comminution on transdermal absorption and rheology of P. cuspidatum ointment (PCO). Methods: Common and ultrafine common powder of P. cuspidatum was got by common and ultrafine comminution technology respectively, and then prepared into common and ultrafine PCO. The skin permeation of common and ultrafine PCO was investigated through in vitro excised mice skin using improved Franz diffusion cell, with polydatin, resveratrol, and emodind as the indexes to compare the effect on the cumulative permeation amount and permeation rate of ointments. Then the effect on the rheology of ointments with apparent viscosity and yield stress as the indexes was compared. Results: The d0.9 of common and ultrafine powder of P. cuspidatum were (210.011 ± 3.468) and (63.496 ± 2.570) μm; And the spans were (5.412 ± 0.055) and (2.913 ± 0.117), respectively. The cumulative permeation amount of polydatin, resveratrol, and emodind of PCO were (2.100 3 ± 0.154 5), (2.114 5 ± 0.341 6), and (6.210 4 ± 0.750 0) μg/cm2, and the permeation rate were (0.175 3 ± 0.022 6), (0.234 0 ± 0.020 2), and (0.337 4 ± 0.051 6) μg/(cm2∙h), respectively. The ultrafine cumulative permeation amounts of polydatin, resveratrol, and emodind of PCO were (14.247 9 ± 4.875 0), (4.399 3 ± 0.628 7), and (6.768 6 ± 0.728 6) μg/cm2, which were 6.8, 2.1, and 1.1 times compared to common PCO; The permeation rates were (0.815 9 ± 0.277 1), (0.313 2 ± 0.043 0), and (0.393 7 ± 0.042 6) μg/(cm2∙h), which were 4.7, 1.3, and 1.2 times compared to common PCO respectively. The cumulative permeation amounts and permeation rates of ultrafine PCO were 6.8, 2.1, 1.1 and 4.7, 1.3, 1.2 times compared to common PCO. The apparent viscosity and yield stress of common and ultrafine PCO were 34.940, 8.865 Pa∙s and 41.211, 7.381 Pa. Conclusion: Compared with common PCO, the transdermal permeability and fineness of ultrafine PCO were improved, and the apparent viscosity and yield stress decreased, which facilitates of spreadability.

Study on the Excretion Pharmacokinetics of Polydatin and Its Metabolite in Beagle Dogs / 中国药房

OBJECTIVE:To study the metabolite and excretion of polydatin (Trans-resveratrol-3-O-glucoside,TRG) and its metabolites in Beagle dog feces,and provide experimental basis for the new drug development of TRG. METHODS:4 Beagle dogs were intragastrically administrated TRG 20％ polyethylene glycol 400 solution 50 mg/kg. LC/MS was used to identify the me-tabolites of TRG in dog feces after 12-24 h of administration,and HPLC was adopted to determine the cumulative excretion rate and excretion speed rate of the major metabolite resveratrol(TR)in feces samples after 0-12,12-24,24-36,36-48,48-72,72-96 h of administration. RESULTS:TR and two sulfate conjugates of TR were found in dog feces,and TR was a major metabolite, while TRG was undetermined. The excretion speed rate of TR in dog feces was the rapidest after 12-24 h of administration,reach-ing 9.65 mg/h. Its cumulative excretion rate was 39.3％,equivalent to 67.2％ of TRG. CONCLUSIONS:After TRG is intragastri-cally administrated to dogs,TRG was mainly excreted in the form of TR via dog feces.

Simultaneous determination of five constituents in Jiangzhi Granules by HPLC / 中成药

AIM To establish an HPLC method for the simultaneous content determination of five constituents in Jiangzhi Granules (Salviae miltiorrhizae Radix et Rhizoma,Polygoni cuspidati Rhizoma et Radix,Artemisiae scopariae Herba,etc.).METHODS The analysis of methanol extract of this drug was performed on a 30 ℃ thermostatic Hanbon-C18 column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrile (A) 0.1％ phosphoric acid (B) flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelengths were set at 286 nm (0-30,40-50 min) and 203 nm (30-40 min).RESULTS Gypenoside A,chlorogenic acid,polydatin,salvianolic acid B and emodin showed good linear relationships within the ranges of 0.176-2.46,0.056-0.84,0.36-5.40,0.192-2.88 and 0.12-1.80 μg (r ＞0.999 0),respectively,whose average recoveries were 97.17％-102.59％ with the RSDs of 1.9％-2.9％.CONCLUSION This accurate and simple method can be used for the quality control of Jiangzhi Granules.

The effect of polydatin on nerve cell apoptosis in hippocampus after ischemia-reperfusion / 国际中医中药杂志

Objective To observe the effect of polydatin on nerve cell apoptosis and the influence of the caspase-8 and FLIP protein expression after ischemia reperfusion injury. Methods Thirty rats were randomly divided into sham operation group, model group and polydatin group. The middle cerebral artery occlusion (MCAO) model of focal cerebral ischemia was established by the method of thread embolism. Rats were subjected to adaptive feeding for the first 7 days and then recieved the treatment for another 10 days. The model of ischemia reperfusion injury was established at the 14th day. The polydatin group received 15 mg/kg of polydatin, and the sham operation group and the model group with the same volume of saline once a day for 15 days. The expression of caspase-8 and FLIP protein in the hippocampus of rats were observed by immunohistochemical staining. The expression of caspase-8 and FLIP protein in the hippocampus of rats were observed by TUNEL method at 72 h after cerebral ischemia-reperfusion. Results Compared with the model group, the number of apoptotic cells of polydatin group significantly decreased in hippocampus CA1 region (P<0.01); The expression of caspase-8 (148.78 ± 6.82 vs. 89.61 ± 7.76) in the polydatin group significantly decreased and the expression of FLIP (127.60 ± 8.52 vs. 150.22 ± 8.53) in the polydatin group was increased significantly in hippocampus CA1 region(P<0.01). Conclusions Polydatin have a protection effect on ischemia reperfusion injury in rats and its mechanism may be inhibition of caspase-8 protein expression, promote the FLIP protein expression.

Study on the Quality Standard of Huoxuesan / 中国药房

OBJECTIVE:To establish the quality standard of Huoxuesan. METHODS:Microscopic identification method was adopted for the qualitative identification of Rhizoma pinelliae,Eupolyphaga seu,Phellodendron amurense and Fructus kochiae in the preparation. HPLC was adopted for the contents determination of phellodendrine,polydatin,jatrorrhizine,berberine and emo-din:the column was ZORBAX Eclipse Plus C18 with methanol-0.1% phosphoric acid(gradient elution)at a flow rate of 0.4 ml/min,the detection wavelength was 240 nm,column temperature was 30 ℃,and the injection volume was 5 μl. RESULTS:Micro-scopic identification map of R. pinelliae,E. seu,P. amurense and F. kochiae was clear. The linear range was 0.159-5.073 μg(r=0.997 4)for phellodendrine、0.149-4.761 μg(r=0.999 9)for polydatin、0.239-7.649 μg(r=0.995 5)for jatrorrhizine、0.165-5.268 μg (r=0.997 2)for berberine、0.012-0.382 μg(r=0.999 9)for emodin;RSDs of precision,stability and reproducibility tests were low-er than 3.0%;recoveries were 98.9%-104.1%(RSD=1.9%,n=6),96.1%-101.7%(RSD=2.5%,n=6),99.6%-105.1%(RSD=2.6%,n=6),90.3%-98.2%(RSD=2.9%,n=6)and 92.9%-96.4%(RSD=2.0%,n=6)respectively. CONCLUSIONS:The es-tablished standard can be used for the quality control of Huoxuesan.

Study on the Improvement Effects and Mechanism of Polydatin on Renal Fibrosis in Rats / 中国药房

OBJECTIVE:To study the improvement effects of polydatin on renal fibrosis in rats and its mechanism. METH-ODS:50 rats was were randomly divided into sham operation group,model group,positive group(benazepril,5 mg/kg)and poly-datin high-dose and low-dose groups(100,50 mg/kg),with 10 rats in each group. Except for sham operation group,renal fibrosis model was induced by unilateral ureter obstruction. After modeling,administration groups were given relevant medicine intragastri-cally,and sham operation group and model group were given 0.5%sodium carboxymethyl cellulose solution once a day for consec-utive 4 weeks. The pathological change of renal tissue was scored. 24 h urinary protein and serum levels of urea nitrogen and creati-nine were determined,and the content of hydroxyproline,mRAN expression of TGF-β1 and FN were detected in renal tissue. RE-SULTS:Compared with sham operation group,pathological score,24 h urinary protein,serum levels of urea nitrogen and creati-nine,the content of hydroxyproline in renal tissue,mRNA expression of TGF-β1 and FN were all increased significantly (P<0.01). Compared with model group,24 h urinary protein,serum levels of urea nitrogen and creatinine and mRNA expression of FN in renal tissue decreased significantly in administation groups;the pathology scores,the content of hydroxyproline in renal tis-sue and mRNA expression of TGF-β1 of positive group and polydatin high-dose group were all decreased significantly(P<0.05 or P<0.01). CONCLUSIONS:Polydatin can prevent kidney fibrosis induced by unilateral ureteral obstruction,the mechanism of which may be associated with the mRNA expression down-regulation of TGF-β1 and FN in renal tissue.

Simultaneous determination of four ingredients in Gan Bawei capsule by HPLC-DAD / 药学实践杂志

Objective To establish an HPLC-DAD method for simultaneous determination of four ingredients in Gan Ba-wei capsule .Methods HPLC method was adopted with C18 column(250 mm × 4.6 mm ,5 μm) ,Acetonitrile as mobile phase A and water as mobile phase B in gradient elution mode . The detection wavelength was 240 nm and the flow rate was 1.0 ml/min .Results Salvianolic acid B reference substance was excellently linear in the range of 15.026~100.17 μg/ml ,the average recovery was 96.6% (n=6)and RSD was 1.8% (n=6) .Paeoniflorin reference substance was excellently linear in the range of 14.335~95.564 μg/ml ,the average recovery was 97.0% (n=6)and RSD was 1.7% (n=6) .Polydatin reference sub-stance was excellently linear in the range of 8.235-54.90 μg/ml ,the average recovery was 97.2% (n=6)and RSD was 1.8%(n=6) .Emodin reference substance was excellently linear in the range of 1.582 5-10.55 μg/ml ,the average recovery was 98.1% (n=6)and RSD was 1.7% (n=6) .Conclusion The method was easy ,accurate ,reproducible ,specific and without in-terference in negative control ,which could be used for quality control of Gan Bawei capsule .