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.

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.

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.

Effect of Ultrafine Grinding on Dissolving-out Quantity of Tanshinone IIA from Radix Salvia Miltiorrhiza

In order to improve the bioavailability, Radix salvia miltiorrhiza raw powder mixed with distilled water （5.5wt%） was ultrafinely ground to nanosize particles using HSCS pulverizer, and the dissolving-out quantity of tanshinone IIA in the filtrate that obtained from nanoparticles suspension and raw powder marinated in water for different time was determined by HPLC. The experimental results show that raw powder can be ultrafinely ground to 133.5 nm at 1500 r/min for 50 min and the molecular structure of active ingredients doesnot change, and the dissolving-out quantity of tanshinone IIA obtained from the filtrate is increased greatly from 12.77 μg/g to 54.55 μg/g.

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.

Effect of Radix Salviae Miltiorrhizae on Production of Lung Oxygen Free Radical Products during Cardiopulmonary Bypass Operation

Objective: The effect of Radix Salviae Miltiorrhizae (RSM) on production of lung oxygen freeradical products during cardiac operations was researched. Methods: Thirty cases of heart valve replacementwere randomly divided into two groups: RSM treatment group (RSM group ) and control group. Blood sampleswere taken from before (right atrium, RA) and after (left atrium, LA) pulmonary circulation before cardiopulmonary bypass operation and 10 minutes, 45 minutes, 2 hours, 6 hours after the aortic crossclamp removal toobserve the level of peroxidation product and the changes in leucocyte count. Results: The number of leucocytesin LA was smaller than that in RA and the amount of peroxidation product in LA was higher than that in RA(P＜0. 01～0. 05 ) during the early phase of lung reperfusion in the control group. There was no significant difference (P＞0. 05) in the amount of peroxidation product and the leucocyte count between LA and RA in theRSM group. Conclusions: RSM could markedly prevent the lung leucocyte aggregation and reduce the production of lung free radical products. It suggested that the effect of RSM of reducing the lung oxygen free radicalproducts was related to its effect of preventing the lung leucocyte aggregation.

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.

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 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.

Chinese Prescription Kangen-karyu as Potential Anti-Alzheimer’s Disease Therapeutic:Analyses of BACE1 and GSK-3βInhibitory Activities

Inhibition ofβ-site amyloid precursor protein-cleaving enzyme 1(BACE1)or glycogen synthase kinase-3β(GSK-3β)is estimated to be the central therapeutic approach for Alzheimer’s disease(AD).In this study,water extract of Kangenkaryu,its crude drug and chemical composition used in oriental medicine were evaluated regarding their BACE1 and GSK-3βinhibitory activities.Fluorescence resonance energy transfer was used to characterize the BACE1 inhibitory effect of Kangen-karyu,its crude drug and chemical composition.GSK-3βactivity was determined using the Kinase-Glo Luminescent Kinase Assay Platform.The water extract of Kangen-karyu inhibited BACE1 and GSK-3βin concentration-dependent manners when compared with reference drugs,quercetin and luteolin.Among six components of Kangen-karyu,the water extracts of Salviae Miltiorrhizae Radix or Cyperi Rhizoma exhibited significant inhibitory effects on BACE1 and GSK-3β.Among the constituents of Salviae Miltiorrhizae Radix extract,salvianolic acid C,salvianolic acid A,rosmarinic acid,and magnesium lithospermate B significantly inhibited BACE1.In addition,they inhibited GSK-3βwith an IC50 value range of 6.97 to 135.35μM.From these results,one of the effectiveness and its mechanisms of action of Kangen-karyu against AD may be the inhibition of BACE1 and GSK-3β,and one of the active ingredients of Kangen-karyu is Salviae Miltiorrhizae Radix and its constituents.

Clinical Study on Early Post-operational Intraperitoneal Chemotherapy and Salviae in Treating Patients of Gastric Cancer

Objective: To evaluate the safety, feasibility and short term efficacy of early post operational intraperitoneal chemotherapy (EPIC) combined with Salviae miltiorrhizae (SM) in treating patients with gastric cancer. Methods: The 136 patients enrolled were divided into 3 groups: the EPIC group, the EPVC group and the control group. The former two groups were treated with SM plus 5 FU started from the second or third day after operation for 5 continuous days by intraperitoneal infusion or intravenous dripping respectively, and the control group was untreated but conventional chemotherapy was given 3 weeks after surgical operation. Toxic and adverse effects of chemotherapy, post operational complications, short term survival rate and intra abdominal tumor recurrence rate were observed and compared.Results: (1) Toxic adverse effects of chemotherapy that occurred in the EPIC group were less than those in the EPVC group significantly ( P <0.05 0.01). (2) Occurrence of serious complications in the EPIC group was not higher than that in the other two groups. (3) The 1 and 2 year survival rate in the EPIC group was higher than those in the other two groups respectively ( P <0.01), while the post operational intra abdominal recurrence rate in EPIC group was significantly lower than that in the other two groups ( P <0.05). Conclusions: Combined therapy of SM and 5 FU in treating patients with gastric cancer by intraperitoneal infusion is not only safe and feasible with mild toxic and side effect, but also produces a more beneficial effects, including less intra abdominal recurrence and satisfactory short term survival rate .

Protecdive Effects of Radix Astrsgalus on Vascular Endotbelial Cells of Patients with Binswanger's Disease

Objective: To compare the effects of Radix Astragalus (RA) on vascular endothelial cells inBinswanger's disease (BD) patients with Radix Salviae miltiorrhizae (RSM). Methods: There were 37 patientswith BD in the treated group and 37 healthy subjects in the control group. Thirty-seven patients were furtherrandomly subdivided into two groups: RA group (19 patients) and RSM group (18 patients). Circulating endothelial cells (CEC) and the levels of endothelin--1 (ET--1 ), nitric oxide (NO) and malondialdehyde (MDA) inthe blood of internal jugular vein which were examined before and after treatments. Results: When comparedwith those of the control group, CEC counts, ET-1 and MDA levels in plasma increased significantly, meanwhile serum NO concentration decreased significantly in the treated group. When compared with those of pretreatment, CEC counts, ET-1 and MDA decreased significantly and serum NO concentration increased significantly after treatment in RA groUp. There were no significant changes of these indices in RSM group after treatment. Conclusions: There are damage and dysfunction of vascular endothelial cells in patients with BD. RA injection is an effective drug to protect vascular endothelial cells of BD patients.

Rapid separation and identification of multiple constituents in Danhong Injection by ultra-high performance liquid chromatography coupled to electrospray ionization quadrupole time-of-flight tandem mass spectrometry

To characterize and identify multiple constituents in Danhong injection(DHI), a fast ultra-high performance liquid chromatography coupled to electrospray ionization quadrupole time-of-flight tandem mass spectrometry(UHPLC-ESI-QTOF/MS) method was established and validated in the present study. A total of 63 compounds, including 33 phenolic acids, 2 C-glycosyl quinochalcones, 6 flavonoid O-glycosides, 4 iridoid glycosides, 6 organic acids, 5 amino acids, and 3 nucleosides, were identified or tentatively characterized. In conclusion, the UHPLC-ESI-QTOF/MS method is useful and efficient for in-depth structural elucidation of chemical compounds in complex matrices of herbal medicines such as DHI.

Therapeutic Effects of Traditional Herbal Medicine on Cerebral Ischemia:A Perspective of Vascular Protection

Although many agents for acute ischemic stroke treatment have been developed from extensive preclinical studies, most have failed in clinical trials. As a result, researchers are seeking other methods or agents based on previous studies. Among the various prospective approaches, vascular protection might be the key for development of therapeutic agents for stroke and for improvements in the efficacy and safety of conventional therapies. Traditional medicines in Asian countries are based on clinical experiences and literature accumulated over thousands of years. To date, many studies have used traditional herbal medicines to prove or develop new agents based on stroke treatments mentioned in traditional medicinal theory or other clinical data. In the current review, we describe the vascular factors related to ischemic brain damage and the herbal medicines that impact these factors, including Salviae Miltiorrhizae Radix, Notoginseng Radix, and Curcumae Rhizoma, based on scientific reports and traditional medical theory. Further, we point out the problems associated with herbal medicines in stroke research and propose better methodologies to address these problems.

Effect of Magnesium Lithosperarnate B on D－galactosamine Induced Liver Injury in Rats

Treated the experimental acute liver injury rat induced by D-galactosamine with magnesiumIithosperamate-B could significantly lower the serum activity of alanine transaminase and aspartate amino-transferase, relieve the necrosis of hepatocytes and elevate the levels of products of arachidonic acid (6-ke-to-prostaglandin 1 , prostaglandin D2 and the total prostaglandin) in liver. These results suggested that mag-nesium lithosperamate B was one of the rnain components of Salvia miltiorrzae in treating liver injury, the ac-tion on elevating the total prostaglandin in liver might be its mechanism.