Progress in research into the mechanism of Radix salviae miltiorrhizae in treatment of acute pancreatitis

BACKGROUND: Acute pancreatitis (AP) is a common acute abdomen with high mortality, multiple complications and complicated causes. There is no effective therapy for AP. Radix salviae miltiorrhizae (Danshen), a traditional herbal medicine, has a low price and a wide range of clinical applications. It is effective to promote blood flow, eliminate stagnancy, and relieve pain. It is also found to be effective in treating AP. We reviewed the progress in research into the mechanism of Radix salviae miltiorrhizae in treatment of AP. DATA SOURCES: An english-language literature search was conducted using MEDLINE (1988-2005) on Radix salviae miltiorrhizae (Danshen) and acute pancreatitis. RESULTS: The mechanisms of Danshen in the treatment of AP include improvement of microcirculatory disturbances; elimination of oxygen free radicals; modulation of the metabolism of lipid inflammatory mediator; and blocking of calcium inflow and prevention of calcium overload.CONCLUSION: Danshen can effectively reduce the mortality and complications of AP.

Experimental Study on the Preventive Mechanism of Salviae Miltiorrhizae Against Atherosclerosis in Rabbits Models

Summary: The preventive mechanism of salviae miltiorrhizae (SM) against experimental atherosclerosis (AS) in rabbits models was investigated. The experimental AS rabbit models were reproduced by feeding the high cholesterol diet. The changes of atherosclerotic plaques in normal group, model group and SM treated group were observed. The levels of serum TG, TC, HDL-C and LDL-C were determined. The immunohistochemistry was used to detect the expression of Bcl-2, Bax and IL-6 proteins in atherosclerotic plaques. The results showed that the level of serum TG in SM treated group was significantly lower than in model group (P<0.01). Immunohistochemistry revealed that the expression of Bcl-2, Bax and IL-6 in model group was significantly higher than in normal group. In the SM group, the expression of Bcl-2 protein was up-regulated and that of Bax was down-regulated. It was suggested that SM could inhibit formation of AS in experimental rabbits. To decrease the expression of Bax and increase the expression of Bcl-2 protein may be one of the mechanisms of SM against atherosclerosis.

Salviae miltiorrhizae ameliorates cirrhosis and portal hypertension by inhibiting nitric oxide in cirrhotic rats

OBJECTIVE: To determine the effect of Salviae miltiorrhizae on cirrhosis and portal hyperiension by inhibiting nitric oxide synthase type Ⅱ (NOSⅡ) in rats. METHODS: Real time RT-PCR was used to detect the expression of NOSII mRNA. The enzymatic activity of nitric oxide synthase and the circulating levels of nitric oxide (NO), systemic and portal hemodynamics, and quantification of cirrhosis were measured with highly sensitive methods. Traditional Chinese medicine was utilized to treat cirrhotic rats and the function of NO was evaluated. Double-blind method was applied in the experiment constantly. RESULTS: The concentration of NO increased markedly at all stages of cirrhosis, and so did the enzymatic activity of NOS, antl the iNOSmRNA expressed greatly. Meanwhile the portal-venous-pressure (PVP), portal-venous-now (PVF) increased significantly. NO, Nos and iNOSmHNA were positively correlated to the quanlity of hepatic fihrosis. Salviae Miltiorrhizae significantly inhibited NO production and inhibited the expression of iNOSmRNA. CONCLUSIONS: The increased hepatic expression of NoSIⅡ is one of the important factors causing cirrhosis and portal hypertension. Salviae Miltiorrhizae significantly ameliorates cirrhosis and portal hypertension.

Effect of salviae miltiorrhizae and ligustrazine hydrochloride injection on axonal regeneration and nerve growth factor expression in a rat model of sciatic nerve injury

BACKGROUND： Studies have shown that both salviae miltiorrhizae and ligustrazine can promote protein expression of nerve growth factor （NGF） and regeneration of peripheral nerve. OBJECTIVE： To verify the effect of salviae miltiorrhizae and ligustrazine hydrochloride injection on axonal regeneration and NGF protein expression in a rat model of sciatic nerve injury. DESIGN, TIME AND SETTING： A randomized, controlled, animal experiment was performed at the Laboratory of Traditional Chinese Medicine and the Institute of Bioengineering of Jinan University from July to December 2008. MATERIALS： Salviae miltiorrhizae and ligustrazine hydrochloride injection （containing 20 mg salviae miJtiorrhizae and 100 mg ligustrazine per 100 mL injection） was provided by Guizhou Baite Pharmaceutical, China; salviae miltiorrhizae and ligustrazine decoctions （containing 1 g raw drug per 1 mL decoction） were provided by Guangzhou Baiyunshan Factory for Traditional Chinese Medicine, China; rabbit-anti-rat NGF monoclonal antibody was provided by Beijing Biosynthesis Biotechnology, China. METHODS： A total of 80 healthy, male, Sprague Dawley rats were used to establish a sciatic nerve injury model via neurotomy, and were then randomly assigned to 4 groups： salviae miltiorrhizae and ligustrazine hydrochloride injection group （intraperitoneal injection of 35 mL/kg per day salviae miltiorrhizae and ligustrazine hydrochloride injection）, saIviae miltiorrhizae group （intragastric peffusion of 2 mL salviae miltiorrhizae）, ligustrazine group （intragastric peffusion of 2 mL ligustrazine）, and model group （intraperitoneal injection of 35 mL/kg per day saline）, with 20 rats in each group. Thereafter, rats in each group were then divided into 4 subgroups according to varying time points of 1, 2, 4, and 8 weeks post-surgery, with 5 rats in each subgroup. MAIN OUTCOME MEASURES： Axons were quantified using chromotrope 2R-brilliant green and silver staining combined with image analysis to calculate the axonal regeneration rate; NGF expression was detected using immunohistochemistry and Western blot analysis; toe interspace was measured by behavior at 4 and 8 weeks. RESULTS： With increasing time after sciatic nerve expression, and toe interspace gradually increased njury, the axonal regeneration rate, NGF protein At 4 and 8 weeks post-surgery, axonal regeneration rate and NGF protein expression were significantly increased in the injured tissue of the salviae miltiorrhizae and ligustrazine hydrochloride injection, salviae miltiorrhizae, and ligustrazine groups, compared with the model group （P 〈 0.05 or P 〈 0.01）, and toe interspace was remarkably enlarged （P 〈 0.05 or P 〈 0.01）, especially in the salviae miltiorrhizae and ligustrazine hydrochloride injection group. CONCLUSION： Salviae miltiorrhizae and ligustrazine hydrochloride injection promoted axonal regeneration and NGF protein expression in the injured sciatic nerve, and also enhanced neurofunctional recovery. Its effect was superior to salviae miltiorrhizae or ligustrazine alone.

Protective effect of ultrashortwave versus radix salviae miltiorrhizae on brains of rats with cerebral ischemia-reperfusion injury

BACKGROUND： HOW to control the effect of oxygen-derived free radicals on development of cerebral injury and cerebral edema is a key factor for treating cerebral ischemia-reperfusion injury. OBJECTIVE： To observe and compare the protective effects, synergistic action and mechanisms of ultrashortwave （USW） and radix salviae miltiorrhizae （RSM） on the focal cerebral ischemia-reperfusion injuries in rats. DESIGN： Randomized controlled animal study SEI-FING： Department of Rehabilitation Medicine, First Hospital affiliated to China Medical University MATERIALS： A total of 160 healthy Wistar rats of both genders and aged 18-20 weeks weighing 250-300 g of clean grade were selected in this study. 5 mL/ampoule RSM injection fluid was produced by the First Pharmaceutical Corporation of Shanghai （batch number： 011019, 0.01 mug）. The USW therapeutic device was produced by Shanghai Electronic Device Factory with the frequency of 40.68 MHz and the maximal export power of 40 W. The first channel of power after modulation was 11 W. METHODS： The experiment was carried out in the Rehabilitation Medicine Department of the First Hospital affiliated to China Medical University from May 2002 to January 2003. Focal ischemia-reperfusion model was established in rats by reversible right middle cerebral artery occlusion with filament. Right cerebral ischemia was for 2 hours and then with 24 hours reperfusion. The scores of neurological deficits were evaluated by 0 to 4 scales. After surgery, 64 successful rats models were divided into four groups according to digital table： control group, USW group, RSM group and RSM ＋ USW group with 16 cases in each group. Rats in control group were intraperitoneally injected with the same volume of saline （0.1 mL/g）; rats in USW group were given small dosage of USW on head for 10 minutes at 6 hours after reperfusion; rats in RSM group were intraperitoneally injected with 0.01 mL/g RSM solution at 30 minutes before reperfusion; rats in RSM ＋ USW group were intraperitoneally injected with 0.01 mL/g RSM parenteral solution at 30 minutes before reperfusion and given small dosage of USW on head for 10 minutes once at 6 hours after reperfusion; sixteen rats in sham operation group did not receive any treatment. All 80 rats were taken brains at 24 hours after reperfusion to measure wet and dry weights to calculate water content： Cerebral water content （%） = （1-dry/wet weight） × 100%. Superoxide dismutase （SOD） activity was measured by hydroxylamine method and malondialdehyde （MDA） content was measured by TBA photometric method. MAIN OUTCOME MEASURES ： Cerebral water content, SOD activity and MDA content RESULTS： All 160 rats except 80 failing in modeling were involved in the final analysis. （① The cerebral water content of left hemisphere made no significant difference （P 〉 0.05）. The cerebral water content of right hemisphere in the control group and the three treatment groups was obviously higher than that of the sham operation group [（81.26±0.77）%, （79.74±0.68）%, （79.76±0.81）%, （79.61±0.79）%, （77.43±0.61）%, P 〈 0.05]. The cerebral water content of right hemisphere in the three treatment groups was obviously lower than that of the control group （P〈 0.05）. There was no significant difference among the three treatment groups （P 〉 0.05）. ② Compared with the control group, SOD activity （right） of the control group decreased obviously （P 〈 0.05）, while MDA content increased obviously （P 〈 0.05）. SOD activity in the three therapeutic groups increased obviously, while MDA content decreased obviously （P 〈 0.05）; there was no significant difference among the three treatment groups （P 〉 0.05）. CONCLUSION： ① USW and RSM therapy have neuroprotective effects against focal cerebral ischemia-reperfusion injuries by means of decreasing cerebral water content and MDA and increasing the activity of SOD. ② Synergistic action was not observed between these two therapeutic methods.

Challenges and strategies in progress of drug delivery system for traditional Chinese medicine Salviae Miltiorrhizae Radix et Rhizoma(Danshen)

Traditional Chinese medicines(TCMs), with a history of thousands of years, are widely used clinically with effective treatment. However, the drug delivery systems(DDSs) for TCMs remains major challenges due to the characteristics of multi-components including alkaloids, flavones, anthraquinones, glycosides,proteins, volatile oils and other types. Therefore, the novel preparations and technology of modern pharmaceutics is introduced to improve TCM therapeutic effects due to instability and low bioavailability of active ingredients. Salviae Miltiorrhizae Radix et Rhizoma, the radix and rhizomes of Salvia miltiorrhiza Bunge(Danshen in Chinese), is a well known Chinese herbal medicine for protecting the cardiovascular system, with active ingredients mainly including lipophilic tanshinones and hydrophilic salvianolic acids.In this review, this drug is taken as an example to present challenges and strategies in progress of DDSs for TCMs. This review would also summary the characteristics of active ingredients in it including physicochemical properties and pharmacological effects. The purpose of this review is to provide inspirations and ideas for the DDSs designed from TCMs by summarizing the advances on DDSs for both singleand multi-component from Danshen.

Study on Effect of IH764-3, an Active Principle of Salviae miltiorrhizae, in Inducing Hepatic Stellate Cell Apoptosis

Objective: To explore the anti fibrotic mechanism of Salviae miltiorrhizae from the view of proliferation and apoptosis of hepatic stellate cells (HSC).Methods: IH764 3, an active principle of Salviae miltiorrhizae, was used to intervene in the cultured HSC in vitro. Cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) method, and the cell apoptosis was examined by electron microscopy, flow cytometer and terminal deoxynucleotidyl transferase mediated dUTP nick end labeling method (TUNEL).Results: MTT showed that IH764 3 has obvious inhibition on the proliferation of HSC. Specific cell apoptosis figures of HSC, such as chromatin agglutination, were seen under electron microscopy in the IH764 3 treated group. By flow cytometer, it was shown that the HSC apoptosis rate in the IH764 3 treated group was higher than that in the control group, and the apoptosis inducing effect of IH764 3 was dose and time dependent. TUNEL analysis showed that the HSC apoptotsis rate was 28.3±1.5% after being incubated for 48 hrs with IH764 3, which was significantly higher than that in the control group (6.7±0.6%, P <0.05).Conclusion: IH764 3 could inhibit the proliferation of HSC and induce its apoptosis. These effects may be one of the anti fibrotic mechanisms of Salviae miltiorrhizae.

EFFECTS OF HYPOXIC ENDOTHELIAL CELL CONDITIONED MEDIUM ON PROLIFERATION AND COLLAGEN SYNTHESIS OF SMOOTH MUSCLE CELLS AND INHIBITORY EFFECTS OF RADIX SALVIAE MILTIORRHIZAE

The effects of hypoxic endothelial cell conditioned medium (HECCM) on proliferation and collagen synthesis of cultured porcine pulmonary arterial smooth muscle cells (PASMCs) were studied by 3H-thymidine (3H-TdR) and 3H-proline incorporations, image analysis for determination of DNA content and colorimetric assay using MTT, and the inhibitory effects of radix salviae miltiorrhizae (RSM) on them were also investigated. The results showed that HECCM could induce enhancement of the enzymatic activity of mitochondria, increase of the nucleic DNA content and increases of the 3H-TdR and 3H-proline incorporations in PASMCs. The 3H-proline incorporation in PASMCs cultured in HECCM was 1.83 times as much as that cultured in normoxic endothelial cell conditioned medium (NECCM). Compared with the control, Chinese herb medicine RSM could inhibit the proliferation of PASMCs cultured in HECCM and decrease the 3H-prolinc incorporation in PASMCs cultured in both HECCM and NECCM (P< 0.001). However, RSM had no ef fects on the nucleic DNA content and 3H-TdR incorporation into DNA of PASMCs cultured in NECCM. It suggests that hypoxia may stimulate the endothelia to synthesize and secrete some cytokines which can stimulate the proliferation and the synthesis of collagen of PASMCs and RSM can inhibit this process.

Experimental Study on Therapeutic Effect of Combined Administration of Radix Salviae Miltiorrhizae. Ligustrazine and Radix Sanchi on Late Hemorrhagic Shock of Rabbits

Objective: To explore the therapeutic mechanism of the combined use of Radix Salviae Miltiorrhizae, ligustrazine and Radix Sanchi in treating late hemorrhagic shock in rabbit. Methods: Rabbit hemorrhagic shock models were set up according to Wiggers' method and treated with the combined therapy. Levels of RBC superoxide dismutase (SOD) activity, plasma malondialdehyde (MDA), blood lactate (BL) and magnesium (Mg++ ) were continuously monitored before shock, 120 minutes after shock, 60 and 120 minutes after reinfusion. Results: (1) Levels of SOD decreased and MDA, AL, Mg++ increased markedly 120 minutes after shock, indicating that there existed cell membrane damage caused by oxygen free radicals in hemorrhagic shock.(2) The combination therapy could alleviate lipid peroxidation injury to tissue, enhance the activity of SOD and lower the concentration of MDA significantly, P < 0. 01 or 0. 05, as compared with the control group. Conclusion: The combined use of the three drugs could gain the same effect with half dosage as that gained from the whole dosage of one single drug. It could also reduce the negative effect of treatment, such as hypotension and slowing down of heart rate

Biomechanical Experimental Study on Effective Fraction of Radix Salviae Miltiorrhizae on Healing of Bone Fracture

Objective: To assess the effect of Danshen 9403 (DS 9403), an effective fraction of Radix Salviae miltiorrhizae (RSM) on healing of bone fracture. Methods: Standardized radial fracture was performed in 120 Wistar rats. The model rats were randomized into four groups: Group A was fed with DS 9403, group B injected with Staphylococcus aureus , group C with normal saline administration, and group D with RSM injection. The treatment began at the first day of fracture. The rats were sacrificed on the day 25, 39 and 50 separately in batches and their intact radii were removed by dissection for detecting load and stress of three point bending test with autograph universal material testing machine (Shimazu, Japan). Results: The parameters of load in DS 9403 treated group on the 39th day and that of stress at 25th, 39th and 50th day were (6.20±1.32)N, (5.71±3.58)N/mm 2, (8.27±2.42)N/mm 2 and (66.25±26.21)N/mm 2 respectively, which were significantly higher than those in other groups, P <0.05. Conclusion: DS 9403 has the action of increasing the strength of fracture broken end.

The Inhibitory Effect of Radix Salviae Miltiorrhizae on Hypoxic Structural Remodeling of Intra－acinarPulmonary Arteries

The inhibitory effect of Radix Salviae Miltiorrhizae (RSM) on hypoxic structural remodeling of intra-acinar pulmonary arteries (IAPA) was observed by light and electron microscopy and morphometry. It was found that RSM can not only dilate IAPA and relieve the hypoxic injuries to endothelia cells,but also inhibit the active muscularization of IAPA in the hypoxic animals, suggesting that RSM plays a very important role in inhibiting structural remodeling of IAPA and pulmonary hypertension.

Treatment of Purpuric Nephritisin Children with Tripterygium Wilfordii Polyglucoside and Radix Salviae Miltiorrhizae

This study deals with the data of Tripterygium Wilfordii polyglucoside (1 mg/kg·day)combined with Radix Salviae Miltiorrhizae (6 - 15 g/d) in the treatment of purpuric nephritis, comparedwith a control group using Tripterygium Wilfordii polyglucoside treatment only. The average time of ede-mad-isappearance and blood pressure-resumption to a normal range were 8. 72 ± 3. 07 and 7. 44 ± 5. 17days in the test group, which was much better than in the control group (P<0. 05) . The average time ofhematuria-disappearance in the test group and the control group were 36. 37 ± 11. 50 and 47. 73 ± 14. 03days respectively. The average time for C3 levels in the urine to resume to normal in the test group and thecontrol group were 40. 33±4. 51 and 50. 00±5. 30 days respectively (P<0.01) . The time for microcircu-lation of nail fold to return to normal after treatment in the test group and the control group were 39. 00±11. 03 and 66. 00± 9. 01 days respectively (P<0. 001) . These results indicate the effect on the test groupwas much better than that on the control group.

Effects of Compound Salviae Miltiorrhizae Injection on Metabolism of Phosphoinosititides in Myocardial Ischemia

Using the model of acute myocardial ischemia in rats formed by ligation of the descending an-terior branch of the left coronary artery, this study showed some effects of compound Salviae miltiorrhizae in-jection ( CSMI) on metabolism of phosphoinositide in myocardial ischemia. The results of the experimentshowed that after 10 minutes of acute ischemia, the function of myocardial messangers ot phosphoinositide in-creased, and the levels of phosphatidylinositol-4, 5-biphosphate ( PIP2 ) and inositol-1, 4, 5-trisphosphate(IP3 ) were obviously higher than those of the non-ischemia control groups ( P < 0. 01 , n = 7) . CSMI (2 glkgbody weight) could markediy inhibit myocardial PIP2 and IP3, whose levels became lower than those of theischemia control groups ( P < 0 . 01 , n = 7 ) .

Effects of Tetramethylpyrazine and Radix Salviae Miltiorrhizae on Collagen Synthesis and Proliferation of Cardiac Fibroblasts

Objective: To explore the effects of Tetramethylpyrazine (TMP) and Radix Salviae Miltiorrhizae (RSM) on collagen synthesis and proliferation of cardiac fibroblasts. Methods: Using collagenase and pancreatin digested rat cardiac tissue assay to isolate cardiac fibroblasts (FB). Different dosage of TMP, RSM and norepinephrine were used to study their effects on the collagen synthesis and proliferation of cultured cardiac FB. Results: Compared with the control group, moderate or high dosage TMP and RSM could significantly inhibit the collagen synthesis and the proliferation of cultured cardiac FB. Moreover, low-dose TMP (50 mg/L) and low-dose RSM (3 g/L) could antagonize the collagen synthesis and the proliferation of cultured cardiac FB stimulated by NE (500μg/L). Conclusion: Both TMP and RSM can inhibit the collagen synthesis and proliferation of cultured cardiac FB processes.The mechanisms of these effects might be correlated to their Ca++ antagonistic action. Original article on CJIM(Chin) 1998; 18(7): 423

Study on Preventive and Therapeutic Effect of Radix Salviae Miltiorrhizae on Recurrence and Metastasis of Liver Cancer

Objective: To study the inhibitory effect of Radix Salviae Miltiorrhizae (RSM) on adhesive and invasive ability of SMMC 7721 liver cancer cells, and on the metastasis and recurrence on post hepatectomy liver cancer in nude mice. Methods: Effect of RSM on SMMC 7721 cell membrane intercellular adhesive molecule 1 (ICAM 1) expression was observed by immunofluorescence flow cytometry; effect on invasive ability and of SMMC 7721 cell and the detachment of which attached to fibronectin (FN) by cell migration experiment; effect on adhesion of SMMC 7721 cell with FN by MTT method and effect on adhesion between 7721 7721, 7721 lymphocyte and 7721 endothelial cell by cell adhesion experiment. LCI D20 human liver cancer metastasis model, after hepatectomy, was used to observe the effect of RSM on recurrence and metastasis of liver carcinoma in nude mice. Results: ICAM 1 expression in SMMC 7721 cells incubated with RSM was significantly lower than that in cells did not treated with RSM. RSM could inhibit the invasive ability of SMMC 7721 cell and made the cells already attached to FN exfoliated. It could also inhibit the adhesion of 7721 7721, 7721 lymphocyte and 7721 endothelial cell. And RSM showed preventive and therapeutic effect on intrahepatic and remote metastasis/recurrence of early and late human post hepatectomy liver cancer in nude mice. Conclusion: RSM could inhibit the invasion and adhesion of SMMC 7721 cell and could also prevent and inhibit metastasis and recurrence of human liver cancer after hepatectomy in nude mice.

Effects of Radix Salviae Miltiorrhizae on Proliferation,Apoptosis and c-myc Protein Expression of Fibroblast in Culture of Kindney with Lupus Nephritis

Objectiv:To observe the effects ofRadix Salviae Miltiorrhizae (SM) on humanfibroblast in culture of kidney with lupus nephritis (LN ). Methods: Fibroblasts wereisolated from culture of kidney biopsy of LN patients, and effect of SM on 3H-TdR incorporated rate of fibroblasts was observed. Theapoptosis and c-myc expression were detectedin the same time by flow cytometry.Results:SM could inhibit the proliferation of fibrolast,and promote the programmed cell deaththrough upregulate c-myc protein expression inhuman renal fibroblasts. Conclusions: Longterm administration of SM in large dosagecould be effective on interstial fibrosis of LN,so that to prevent or reduce the scar tissue for-mation and teatrd the occurrence of uremia.

Diterpenoid Tanshinones,the extract from Danshen(Radix Salviae Miltiorrhizae)induced apoptosis in nine human cancer cell lines

OBJECTIVE:To identify the active anti-tumor constituents in the extract from Danshen(Radix Salviae Miltiorrhizae) and investigate the mechanisms underlying the actions.METHODS:First,we introduced a two-step counter-current chromatography to extract the therapeutically active diterpenoid,tanshinone from Danshen(Radix Salviae Miltiorrhizae).The cholecystokinin(CCK-8) method was used to evaluate the inhibitory effect of diterpenoid tanshinone in liver cancer QGY-7703,lung cancer PC9,lung cancer A549,gastric cancer MKN-45,gastric cancer HGC-27,colon cancer HCT116,myeloma cell U266/RPMI8226,and human breast cancer MCF-7 in vitro.Fluorescence staining was used to observe the cytotoxicity ofditerpenoid tanshinone on PC9 cells.The Western blot was used to detect apoptosis-related protein poly ADP-ribose polymerase(PARP),cysteinyl aspartate specific proteinase3/9(caspase3/9),and cleaved-cysteinyl aspartate specific proteinase3/9(cleaved-caspase3/9).The endoplasmic reticulum stress-related activating transcription factor 4(ATF4),phosphorylated eukaryotic initiation factor 2α(p-e IF2α),and phosphorylated jun amino-terminal kinase(p-JNK),and caspase-12 were also analyzed using the Western blot.RESULTS:Diterpenoid tanshinone inhibited the nine human tumor cell lines,with an IC50 of4.37-29 μg/m L,with the PC9 and MCF-7 displaying the lowest values.Fluorescence staining showed a lethal effect of diterpenoid tanshinone on PC9 cells.The Western blot showed that the expression of caspase3/9 protein and ATF-4 protein decreased gradually.However,the PARP,cleaved-caspase 3/9and the expression of p-e IF2 α,P-JNK,and caspase-12 increased gradually,in a dose-dependent fashion.CONCLUSION:We successfully introduced a two-step counter-current chromatography method to extract diterpenoid tanshinone,and demonstrated its antitumor activity.Diterpenoid tanshinone can induce apoptosis in nine human cancer cell lines.

The Effect of Soil Enzymes and Polysaccharides Secreted by the Roots of Salvia miltiorrhiza Bunge under Drought,High Temperature,and Nitrogen and Phosphorus Deficits

Root exudates serve as crucial mediators for information exchange between plants and soil,and are an important evolutionary mechanism for plants’adaptation to environmental changes.In this study,15 different abiotic stress models were established using various stress factors,including drought(D),high temperature(T),nitrogen deficiency(N),phosphorus deficiency(P),and their combinations.We investigated their effects on the seedling growth of Salvia miltiorrhiza Bunge and the activities of Solid-Urease(S-UE),Solid-Nitrite Reductase(S-NiR),Solid-Nitrate Reductase(S-NR),Solid-Phosphotransferase(S-PT),and Solid-Catalase(S-CAT),as well as the contents of polysaccharides in the culture medium.The results showed that the growth of S.miltiorrhiza was inhibited under 15 stress conditions.Among them,13 stress conditions increased the root-shoot ratio.These 15 stress conditions significantly reduced the activity of S-NR,two combinations significantly improved the activity of S-NIR,they were synergistic stresses of high temperature and nitrogen deficiency(TN),and synergistic stresses of drought and nitrogen deficiency(DN)(p<0.05).The activity of S-UE was significantly improved under N,D,T,synergistic stresses of drought and high temperature(DT),DN,synergistic stresses of drought and phosphorus deficiency(DP),and synergistic stresses of high temperature,nitrogen,and phosphorus deficiency(TNP)stress conditions(p<0.05).Most stress combinations reduced the activity of S-PT,but D and T significantly improved it.(p<0.05).The N,DN,and TN stress conditions significantly reduced S-CAT activity.The P,DT,and synergistic stresses of drought,high temperature,and phosphorus deficiency(DTP)significantly decreased the total polysaccharide content of the soil(p<0.05).The research suggested that abiotic stress hindered the growth of S.miltiorrhiza and altered the behavior of root secretion.Roots regulated the secretion of several substances in response to various abiotic stresses,including soil nitrogen cycle enzymes,phosphorus transport-related enzymes,and antioxidant enzymes.In conclusion,plants regulate the utilization of rhizosphere substances in response to abiotic stresses by modulating the exudation of soil enzymes and polysaccharides by the root system.At the same time,soil carbon sequestration was affected by the adverse environment,which restricted the input of organic matter into the soil.

Study on Optimization of Ethanol Reflux Extraction of Phenolic Acids from Salvia miltiorrhiza

The extraction technology of phenolic acid compounds from Salvia miltiorrhiza by ethanol reflux was studied. In this experiment, salvianolic acid B standard was used to make the standard curve. Single factor experiment and orthogonal experiment were used to study the extraction of different ethanol concentrations, reflux times and material-to-liquid ratios. The OD value of salvianolic acid compounds was measured with a spectrophotometer. The extraction rate of phenolic acid compounds under different extraction conditions was calculated through a regression equation, so as to obtain the optimal conditions for the ethanol reflux extraction process of Salvia miltiorrhiza. The experimental data can provide a reference for the ethanol reflux extraction process of salvianolic acids in the industry. According to the experiment, the extraction rate of phenolic acids in Salvia miltiorrhiza was the highest when the ethanol concentration was 60%, the reflux time was 1.5 hours, and the ratio of material-to-liquid was 1:10.