Salprzesides A and B: two novel icetexane diterpenes with antiangiogenic activity from Salvia przewalskii Maxim.

Two novel icetexane diterpenes were isolated from Salvia przewalskii Maxim., namely Salprzesides A (1) and B (2), together with two known abietane-type diterpenes respectively identified as sahandinone (3) and miltirone (4). The structures of isolated compounds were determined by UV, IR, HR-ESI-MS, 1D and 2D NMR analysis. The in vitro antiangiogenic activities of compounds 1-4 were studied against human umbilical vascular endothelial cells (HUVECs). The IC50 values of compounds 1-4 ranged from 4.22 ± 1.07 to 39.31 ± 2.17 µM against HUVECs.

Regioisomers Salviprolin A and B, Unprecedented Rosmarinic Acid Conjugated Dinorditerpenoids from Salvia przewalskii Maxim.

Salvia przewalskii Maxim is a perennial plant from the genus Salvia (family Lamiaceae). The roots of S. przewalskii were long used as a traditional herb to treat blood circulation related illnesses in China. As part of our continuing interest in polycyclic natural products from medicinal plants, two unprecedented adducts comprised of a dinor-diterpenoid and a 9'-nor-rosmarinic acid derivative, linked by a 1,4-benzodioxane motif (1 and 2), were isolated from the roots of S. przewalskii. Their structures were established by extensive spectroscopic approaches including 1D, 2D NMR, and HRFABMS. Their cytotoxic activities against five human tumor cell lines were evaluated.

Increased phenolic acid and tanshinone production and transcriptional responses of biosynthetic genes in hairy root cultures of Salvia przewalskii Maxim. treated with methyl jasmonate and salicylic acid.

The purpose of this study is to reveal the impact of the plant hormone salicylic acid (SA) and methyl jasmonate (MeJA) on the growth, effective components accumulation, and related gene expression of the hairy root of Salvia przewalskii Maxim. Various concentrations of SA (0, 25, 50, 100, 200 µM) or MeJA (0, 50, 100, 200, 400, 600 µM) were added to the culture medium of Salvia przewalskii Maxim. Low concentrations of SA promoted the growth of hairy root, while a high concentration inhibited it. 0 to 400 µM MeJA promoted the growth of hairy root, but 600 µM MeJA starts to inhibit its growth. 50 µM SA and 400 µM MeJA significantly enhanced the production of caffeic acid, rosmarinic acid, salvianolic acid B, cryptotanshinone, and tanshinone IIA. In general, 50 µM SA can be used to accumulate of tanshinone in hairy roots of S. przewalskii with 6 days. 400 µM MeJA can be used to accumulate of phenolic acids in hairy roots of S. przewalskii with 3 days. The selected genes in the tanshinone and phenolic acid biosynthetic pathway were upregulated with elicitation. To obtain a higher yield and content of secondary metabolites, it is advisable to use 50 µM SA or 400 µM MeJA as the optimal doses to cultivate the hairy root of S. przewalskii. This study provides, for the first time, an efficient tanshinone and phenolic acid production method for S. przewalskii.

Bioactive constituents of Salvia przewalskii and the molecular mechanism of its antihypoxia effects determined using quantitative proteomics.

Context: Environmental hypobaric hypoxia induces several physiological or pathological responses in individuals in high-altitude regions. Salvia przewalskii Maxim (Labiatae) (SPM) is a traditional Chinese herbal medicine and has known antibacterial, antiviral, antioxidant, anti-thrombotic, and anti-depressant activities.Objective: This study examined the antihypoxia effects of SPM in vivo.Materials and methods: The dried and pulverised of SPM was extracted from root crude drug with 70% ethanol with ultrasound. Male Sprague-Dawley rats were divided into three groups (n = 10): normal group, hypoxia group (altitude of 4260 m), and hypoxia + SPM group (altitude of 4260 m, SPM of 1.0 g/kg/day). The experiment persisted for 4 weeks. The mean pulmonary arterial pressure (mPAP), hypoxia-inducible factor-1α (HIF-1α) mRNA, and lung pathology were analysed using pulmonary artery pressure recorder, quantitative polymerase chain reaction, and histopathological analysis. Moreover, the effects of SPM on lung proteomes during hypoxia were observed by a TMT-based proteomic approach.Results: Pre-treatment with SPM decreased mPAP (24.86%) and HIF-1α (31.24%), and attenuated the pathological changes in lung tissues. In addition, a total of 28 proteins were differentially expressed in lung of hypoxia + SPM group (fold change > ± 1.2 and p < 0.05). The differentially altered proteins were primarily associated with antioxidative stress, as evidenced by the downregulated expression of Adh7, Cyp2d1, Plod2, Selenow, ND3, and Fabp1, and fructose metabolism, as evidenced by the downregulated expression of Khk and Aldob.Discussion and conclusions: These results suggested that SPM is a promising drug for antihypoxia. The mechanism of action might be related to increasing antioxidant capacity and inhibiting fructose metabolism.

Extract of Salvia przewalskii Repair Tissue Damage in Chronic Hypoxia Maybe through the RhoA-ROCK Signalling Pathway.

Salvia przewalskii Maxim is a traditional Chinese herbal medicine and is known to have antibacterial, antiviral, anti-oxidant, anti-thrombotic and anti-depressant properties. However, the major active components of S. przewalskii and its anti-hypoxic effects are still unclear. This study probed the major active component and anti-hypoxic activity of S. przewalskii. The major active components of S. przewalskii were detected by HPLC. The anti-hypoxic effects of S. przewalskii were detected in mice and a rat model of hypoxic preconditioning. The results showed that there are eight active components, including sodium danshensu, rosmarinic acid, lithospermic acid, salvianolic acid B, dihydrotanshinone I, cryptotanshinone, tanshinone I and tanshinone IIA, and each component showed a certain anti-hypoxic effect. Moreover, S. przewalskii enhanced anti-hypoxia in mice, which was manifested as prolonged survival time in acute hypoxic preconditioning and the amelioration of acute hypoxia-induced changes in the activity of superoxide dismutase (SOD) and lactate dehydrogenase (LDH). In addition, S. przewalskii also repaired tissue damage in chronic hypoxia by downregulating hypoxia inducible factor-1α (HIF-1α), proliferating cell nuclear antigen (PCNA), Bcl-2, CDK4, CyclinD1 and P27Kip1 and inhibiting pro-inflammatory cytokines and the RhoA-Rho-associated protein kinase (ROCK) signalling pathway. Our findings provide new insight into the anti-hypoxic effect of S. przewalskii as a promising agent for high-altitude pulmonary hypertension treatment.

Salvia przewalskii extract of total phenolic acids inhibit TLR4 signaling activation in podocyte injury induced by puromycin aminonucleoside in vitro.

BACKGROUND: TLR4 signaling is known to be involved in podocyte injury. We have previously shown that Salvia przewalskii extract of total phenolic acids (SPE) and its active monomer salvianolic acid B (SalB) and rosmarinic acid (RA) protect podocytes from injury induced by PAN. In the present study, we test whether SPE inhibits TLR4 signaling. METHODS: The conditionally immortalized mouse podocytes were treated with SPE, SalB, RA, SalB + RA or tacrolimus for 30 min, followed by PAN (100 µg/mL) for 24 h. The F-actin staining with phalloidin was used to assess cytoskeletal injury in the podocytes. Western blotting and semi-quantitatives RT-PCR were used to assess the changes of the components in the TLR4 signaling pathway. RESULTS: (1) The F-actin stress fibers of podocytes were almost completely disrupted after PAN treatment for 24 h, and the disruption was significantly alleviated by SPE; (2) the PAN-induced elevation of mRNA levels of TLR4, MyD88 and p65 were inhibited except p65 with high-dose SalB; (3) consistently, the protein levels of TLR4, MyD88 and pp65 were significantly elevated by PAN, and SPE, SalB, RA and admixture, respectively, attenuated the elevations of TLR4 and pp65 proteins; (4) SPE and tacrolimus have a similarly strong effect on inhibition of the expression of TLR4 signaling components. CONCLUSIONS: SPE protects podocytes from PAN-induced injury at least partly through inhibiting TLR4 signaling. SPE is as strong as tacrolimus in inhibiting TLR4 signaling in podocytes.

The effects of Salvia przewalskii total phenolic acid extract on immune complex glomerulonephritis.

CONTEXT: Salvia przewalskii Maxim. (Lamiaceae) is a Chinese herbal medicine that has long been used for the treatment of cardiovascular disease. OBJECTIVE: The study investigated the therapeutic efficacy of S. przewalskii total phenolic acid extract (SPE) on immune complex glomerulonephritis (ICG) in rats. MATERIALS AND METHODS: Sixty-two Wistar rats were randomized into six groups. ICG was induced in all groups except normal control group. SPE was administered intragastrically at 24 h intervals for 40 consecutive days. Urine protein (UP), total serum protein (TSP), serum albumin (SA), serum cholesterol (SC) and serum urea nitrogen (SUN) were measured one day before, on day 20 and 40 after SPE administration. On day 40 after SPE administration, the kidneys were removed and prepared into pathologic sections. In addition, kidney wet mass was measured for calculating the kidney wet mass coefficient (KWMC). RESULTS: UP excretion was reduced significantly on day 20 after SPE administration in all three SPE groups as compared with that in medium group, and this effect was observable continuously until 40 days after SPE administration. Compared with medium group, TSP and SA were increased in all three SPE groups after 40 days treatment, while SC and SUN were decreased. KWMC was decreased significantly in 100 mg/kg SPE group after 40 days treatment compared with that in medium group. Histopathologic analyses showed that renal inflammatory infiltration and kidney intumesce were alleviated in all three SPE groups. CONCLUSIONS: SPE may be a potential therapeutic drug for glomerulonephritis.

Nectar replenishment maintains the neutral effects of nectar robbing on female reproductive success of Salvia przewalskii (Lamiaceae), a plant pollinated and robbed by bumble bees.

Background and Aims: It has been suggested that the dynamics of nectar replenishment could differ for flowers after being nectar robbed or visited legitimately, but further experimental work is needed to investigate this hypothesis. This study aimed to assess the role of nectar replenishment in mediating the effects of nectar robbing on pollinator behaviour and plant reproduction. Methods: Plant-robber-pollinator interactions in an alpine plant, Salvia przewalskii , were studied. It is pollinated by long-tongued Bombus religiosus and short-tongued B. friseanus , but robbed by B. friseanus . Nectar production rates for flowers after they were either robbed or legitimately visited were compared, and three levels of nectar robbing were created to detect the effects of nectar robbing on pollinator behaviour and plant reproduction. Key Results: Nectar replenishment did not differ between flowers that had been robbed or legitimately visited. Neither fruit set nor seed set was significantly affected by nectar robbing. In addition, nectar robbing did not significantly affect visitation rate, flowers visited within a plant per foraging bout, or flower handling time of the legitimate pollinators. However, a tendency for a decrease in relative abundance of the pollinator B. religiosus with an increase of nectar robbing was found. Conclusions: Nectar robbing did not affect female reproductive success because nectar replenishment ensures that pollinators maintain their visiting activity to nectar-robbed flowers. Nectar replenishment might be a defence mechanism against nectar robbing to enhance reproductive fitness by maintaining attractiveness to pollinators. Further studies are needed to reveal the potential for interference competition among bumble bees foraging as robbers and legitimate visitors, and to investigate variation of nectar robbing in communities with different bumble bee species composition.

Salvia przewalskii extract of total phenolic acids against puromycin aminonucleoside-induced oxidative stress in podocytes in vitro and in vivo / 第二军医大学学报

Objective To determine the effect of Salvia przewalskii extract of total phenolic acids (SPE) on puromycin aminonucleoside (PAN)-induced oxidative stress in podocytes of rats in vivo and the effect of SPE on PAN-induced oxidative stress in podocytes of mice in vitro. Methods (1) Nephropathy rat model was established by PAN and was given intervention with SPE and tacrolimus. The renal tissue samples were obtained for WT1 staining to calculate the number of podocytes on the 5th# 10th# 15th and 21st day. The intensities of 8-hydroxy-27-deoxyguanine (8-OHdG) were evaluated by immunofluorescence. (2)The podocytes of mice were exposed to PAN for 24 h in vitro#, and then SPE# salvianolic acid B (SalB) # rosmarinic acid (RA) or tacrolimus were added for 6# 12# 24# and 48 h culture. Then the cytoskeleton distribution of podocytes, indicated by F-actin# was observed by fluorescence microscopy, and the intracellular reactive oxygen species (ROS) production was measured by flow cytometry. Results (1) Decrease of podocytes per glomerular volume as measured by counting WT1-positive cells was started on day 5 in each group except normal control (NC) group# and on day 15 glomerular podocytes in PAN group was significantly less than that in the NC group ([14. 4 + 0. 7]/glomerular volume vs [37. 2 + 1. 5]/glomerular volume# P<0. 05). The numbers of glomerular podocytes in SPE group and positive group (tacrolimus group) were more than that in PAN group at all time points. The glomerular podocyte count of high-dose SPE group was similar to that of positive group on day 15 ([21. 7 + 1. 0]/glomerular volume vs [23. 6 + 1. 2]/glomerular volume# P<0. 05). After injection of PAN# 8-OHdG intensities were increased in each group except normal control group on day 5; and the intensities peaked on day 10 and then began to decrease# but still higher than that of the normal control group on day 15. The intensities of 8-OHdG in renal tissue was decreased after intervention# and those of the tacrolimus and high-dose SPE groups were similar. (2) In vitro study found that F-actin of podocytes was almost completely disrupted 24 h after PAN treatment# with disrupted filamentous structure. After the treatment with tacrolimus, SPE, SalB and RA# the PAN induced injury of podocytes was lessened# with reappeared polarity distribution of intracellular microfilaments. Compared with NC group# the ROS production in podocytes was significantly increased in PAN group (P<0. 05). After treatment of podocyte with drugs# the ROS production was decreased. The cellular ROS production of positive control group was similar to those in tacrolimus group, low-dose SPE group, high-dose SalB group and RA group at 24 h. Compared with RA,SalB had a better efficacy in reducing ROS# and the reducing effect had a positive relation with drug dose. Conclusion Our study suggests that SPE can protect podocytes from PAN-induced oxidant stress in vivo and in vitro.

Salviprzols A and B, C21- and C22-terpenoids from the roots of Salvia przewalskii Maxim.

Two C21- and C22-terpenoids, salviprzols A (1) and B (2), together with 24 known compounds including 17 diterpenoids (3-19), a triterpenoid (20), and 6 phenolic derivatives (21-26), were isolated from the roots of Salvia przewalskii Maxim. Salviprzols A and B represented a new subtype of C23-terpenoids featured by an additional 2-oxopropyl moiety at C-12 and a rare γ-hydroxyl-α-methyl-α,ß-unsaturated-γ-lactone ring system. Their structures were elucidated by extensive spectroscopic analyses, and the structure of 2 was confirmed by a single-crystal X-ray diffraction crystallography. The cytotoxic activities of the new isolates were tested. A plausible biosynthetic pathway for 1 and 2 was also proposed.

Induction, proliferation, and differentiation on callus culture of Salvia przewalskii / 中草药

Objective: To investigate the effects of different concentrations of hormonal combinations on induction, proliferation, and differentiation in Salvia przewalskii callus culture. Methods: Using sterile leaves of S. przewalskii as expiants, the different concentrations of 2, 4-D, NAA, 6-BA, and KT on induction, proliferation of callus were optimized by orthogonal test. Meanwhile the suitable hormonal combinations for differentiation of callus were studied. Results: The results showed that taking the inductive rate and the relative growth rate as norms, 6-BA and 2, 4-D had the most significant effects on the growth of cell division in S. przewalskii; The best hormonal combination for induction in S. przewalskii callus was 2, 4-D 0.5 mg/L + NAA 1.0 mg/L + 6-BA 0.5 mg/L and the best hormonal combination for proliferation in S. przewalskii callus was 2, 4-D 0.5 mg/L + 6-BA 2.0 mg/L + KT 0.1 mg/L. The medium of MS + NAA 0.5 mg/L + 6-BA 1.0 mg/L was beneficial to the differentiation of adventitious buds, and 1/2 MS + IBA 1.0 mg/L was helpful for the differentiation of adventitious roots. Conclusion: The system of organization culture which is established in this study is suitable for the regeneration of S. przewalskii.

Comparison of energy metabolism on brain lacking oxygen between Salvia miltiorrhiza Bunge injection and Salvia przewalskii Maxim injection in mice / 中国临床药理学与治疗学

AIM: To compare the influence of salvia miltiorrhiza Bunge njection (SMBI) and salvia przewalskii Maxim injec tion (SPMI) on energy metabolism of acute brain lacking oxygen. METHODS : The whole brain lacking oxygen model was made by injecting NaNO 2 in mice and then wrote down the existent time of mice and detected the contents of Na +-K +-ATPase, Ca 2+ -ATPase and lactic acid (LD) in the brain. RESULTS: SMBI and SPMI both significantly lengthened the existent tim e of mice with the whole brain lacking oxygen, decreased the contents of LD and increased the activity of ATPase in mice. In increasing the activity of ATPase, the influence of SPMI was more than that of SMBI (P 0.05 ). CONCLUSION: SMBI an d SPMI have distinctive protection on energy metabolism of acute brain lacking o xygen. Both of them can lengthen the existent of mice, reduce the contents of LD in brain and increase the activity of ATPase, but SPMI is more excellent than S MBI in increasing the activity of ATPase.