Schisandrin B Protects against Ischemic Brain Damage by Regulating PI3K/AKT Signaling in Rats / 中国结合医学杂志

OBJECTIVE@#To explore the effect and mechanism of schisandrin B (Sch B) in the treatment of cerebral ischemia in rats.@*METHODS@#The cerebral ischemia models were induced by middle cerebral artery occlusion (MCAO) and reperfusion. Sprague-Dawley rats were divided into 6 groups using a random number table, including sham, MCAO, MCAO+Sch B (50 mg/kg), MCAO+Sch B (100 mg/kg), MCAO+Sch B (100 mg/kg)+LY294002, and MCAO+Sch B (100 mg/kg)+wortmannin groups. The effects of Sch B on pathological indicators, including neurological deficit scores, cerebral infarct volume, and brain edema, were subsequently studied. Tissue apoptosis was identified by terminal transferase-mediated dUTP nick end-labeling (TUNEL) staining. The protein expressions involved in apoptosis, inflammation response and oxidative stress were examined by immunofluorescent staining, biochemical analysis and Western blot analysis, respectively. The effect of Sch B on phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling was also explored.@*RESULTS@#Sch B treatment decreased neurological deficit scores, cerebral water content, and infarct volume in MCAO rats (P<0.05 or P<0.01). Neuronal nuclei and TUNEL staining indicated that Sch B also reduced apoptosis in brain tissues, as well as the Bax/Bcl-2 ratio and caspase-3 expression (P<0.01). Sch B regulated the production of myeloperoxidase, malondialdehyde, nitric oxide and superoxide dismutase, as well as the release of cytokine interleukin (IL)-1 β and IL-18, in MCAO rats (P<0.05 or P<0.01). Sch B promoted the phosphorylation of PI3K and AKT. Blocking the PI3K/AKT signaling pathway with LY294002 or wortmannin reduced the protective effect of Sch B against cerebral ischemia (P<0.05 or P<0.01).@*CONCLUSIONS@#Sch B reduced apoptosis, inflammatory response, and oxidative stress of MCAO rats by modulating the PI3K/AKT pathway. Sch B had a potential for treating cerebral ischemia.

Network analysis and experimental verification of Schisandrin B reduces intestinal ischemia reperfusion injury / 中国临床药理学与治疗学

AIM: To explore schisandrin B (Sch B) pretreatment reduces intestinal ischemia reperfusion injury (IIRI) through inhibiting apoptosis by activation of Nrf2/HO-1 signing pathway in mice by network pharmacology and in vivo experiment. METHODS: (1) The targets of Sch B and IIRI were searched from online databases, Drawing Venn diagram to obtain the common target of them. Cytoscape software was imported to construct the protein-protein interaction (PPI) network to establish the "Drugs-Disease-core target gene" network. The mechanism of Sch B against IIRI was predicted through GO and KEGG enrichment analysis. (2) Thirty-six C57BL/6J mice were randomly divided into six groups (n = 6). The model of IIRI was established in four groups except the sham operation group. Three of the groups were pretreated with Sch B, Nrf2 inhibitor ML385, and Sch B + ML385, respectively. After the experiment, intestinal tissue samples were taken for HE staining, Chiu ' s score, apoptosis staining, immunohistochemistry (IHC), and immunoblotting (Western blot). RESULTS: A total of 412 Sch B related tar- gets, 2 166 IIRI related targets and 153 common targets were screened out through network pharmacology. There were 88 "Sch B-IIRI-core target gene" included NFE2L2 (Nrf2), HMOX1 (HO-1), BCL2, CASP3 (caspase 3), and so on. KEGG enrichment analysis screened 163 related pathways, apoptosis pathway ranked high showing that the pathway may play a key role in the treatment of IIRI by Sch B. The animal experiment had shown that Sch B reduced the Chiu's score and apoptotic while upregulating Nrf2, HO-1, Bcl-2 protein expression levels and Bcl-2/Bax, downregulating Bax, and cleaved caspase-3 expression levels, thereby reducing IIRI in mice, and that Nrf2 inhibitor ML385 reversed this process (P < 0.05). CONCLUSION: This study reveals that Sch B has the characteristics of multi-target and multi-pathway in the reduction of IIRI, and Sch B can reduce IIRI through inhibiting apoptosis by activation of Nrf2/ HO-1 pathway.

Experimental study of schisandrin B attenuates acute myocardial ischemia injury in rats by inhibiting cardiomyocyte apoptosis / 中国临床药理学与治疗学

AIM: To explore the protective effect and mechanism of schisandrin B (Sch B) on myocardial ischemia-reperfusion injury (MIRI) in rats. METHODS: Forty SD rats were randomly divided into control group, MIRI group, MIRI +30 mg/kg Sch B (MIRI + L-Sch B) group and MIRI +60 mg/kg Sch B (MIRI + H-Sch B) group, ten in each group, Sch B were intragastrically for 7 days. Ligation of the left anterior descending branch was used to establish MIRI model, control group rats only underwent sham operation. The ardiac function parameters of ejection fraction (EF), fractional shortening (FS), maximum rate of increase in left ventricular pressure (+ dp/dt

Schisandrin B Inhibits NLRP3 Inflammasome Pathway and Attenuates Early Brain Injury in Rats of Subarachnoid Hemorrhage / 中国结合医学杂志

OBJECTIVE@#To determine whether Schisandrin B (Sch B) attenuates early brain injury (EBI) in rats with subarachnoid hemorrhage (SAH).@*METHODS@#Sprague-Dawley rats were divided into sham (sham operation), SAH, SAH+vehicle, and SAH+Sch B groups using a random number table. Rats underwent SAH by endovascular perforation and received Sch B (100 mg/kg) or normal saline after 2 and 12 h of SAH. SAH grading, neurological scores, brain water content, Evan's blue extravasation, and terminal transferase-mediated dUTP nick end-labeling (TUNEL) staining were carried out 24 h after SAH. Immunofluorescent staining was performed to detect the expressions of ionized calcium binding adapter molecule 1 (Iba-1) and myeloperoxidase (MPO) in the rat brain, while the expressions of B-cell lymphoma 2 (Bcl-2), Bax, Caspase-3, nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3), apoptosis-associated specklike protein containing the caspase-1 activator domain (ASC), Caspase-1, interleukin (IL)-1β, and IL-18 in the rat brains were detected by Western blot.@*RESULTS@#Compared with the SAH group, Sch B significantly improved the neurological function, reduced brain water content, Evan's blue content, and apoptotic cells number in the brain of rats (P<0.05 or P<0.01). Moreover, Sch B decreased SAH-induced expressions of Iba-1 and MPO (P<0.01). SAH caused the elevated expressions of Bax, Caspase-3, NLRP3, ASC, Caspase-1, IL-1β, and IL-18 in the rat brain (P<0.01), all of which were inhibited by Sch B (P<0.01). In addition, Sch B increased the Bcl-2 expression (P<0.01).@*CONCLUSION@#Sch B attenuated SAH-induced EBI, which might be associated with the inhibition of neuroinflammation, neuronal apoptosis, and the NLRP3 inflammatory signaling pathway.

Schisandrin B induces apoptosis of human breast cancer MDA-MB-231 cells through ROS mediated endoplasmic reticulum stress / 药学实践杂志

Objective To study the effects of schisandrin B (Sch B) on the apoptosis of human breast cancer MDA-MB-231 cells and its mechanism. Methods Cell counting reagent (CCK-8) was used to detect the effect of Sch B on the survival rate of MDA-MB-231 cells. MDA-MB-231 cells were treated with Sch B (10, 20, 40 μmol/L) for 24 hours. The cell death was detected by Annexin V-FITC/PI. The levels of intracellular reactive oxygen species (ROS) were detected by DCFA-DA fluorescent probe. Apoptosis and the expression of endoplasmic reticulum stress related proteins (Bcl-2、Bax、CHOP、GPR78、PERK、p-PERK、p-eIF2α、eIF2) were detected by Western blot. Results Compared with the blank group, the cell survival rate decreased significantly (P<0.01) with the increase of Sch B concentration, and its IC50 was 19.16 μmol/L. Compared with the control group, Sch B groups (10, 20, 40 μmol/L) inhibited cell clone formation in a dose-dependent manner (P<0.05). Sch B groups (10, 20, 40 μmol/L) induced apoptosis (P<0.05), significantly reduced the expression of anti-apoptotic protein Bcl-2 and significantly increased the expression of pro-apoptotic protein Bax (P<0.05). Sch B groups (10, 20, 40 μmol/L) significantly increased the level of intracellular ROS in a dose-dependent manner (P<0.05). Sch B groups (10, 20, 40 μmol/L) stimulated endoplasmic reticulum stress and increased the expressions of endoplasmic reticulum stress-related proteins CHOP, GPR78 and p-eIF2α in a dose-dependent manner (P<0.05). Conclusion Sch B induces apoptosis of MDA-MB-231 cells through ROS mediated endoplasmic reticulum stress.

Prescription Technology Optimization of RPV Modified Paclitaxel and Schisandrin B Liposomes and Preliminary Evaluation of Antitumor Activity in vitro / 中国药房

OBJECTIVE：To prepare paclit axel and schisandrin B liposomes modified by cell penetrating peptide RPV ，and to preliminarily evaluate its anti-tumor activity in vitro . METHODS ：RPV modified paclitaxel and schisandrin B liposomes were prepared by film dispersion method. Box-Benhken design-response surface methodology was used to optimize the prescription technology of RPV modified paclitaxel and schisandrin B liposomes using the amount of cholesterol and paclitaxel ，the time interval of ultrasound probe as factors ，average entrapment efficiency of paclitaxel and schisandrin B was used as the index. The liposomes prepared by the optimal technology were characterized. Sulfonylrhodamine B staining method was used to investigate in vitro toxicity of RPV modified blank liposomes ，paclitaxel and schisandrin B liposomes ，RPV modified paclitaxel and schisandrin B liposomes to human ovarian cancer cell SK-OV- 3. The effects of 3 kinds of liposomes on the migration and invasion ability of SK-OV-3 cells were investigated by cell scratch test and Transwell chamber invasion test. RESULTS ：The optimal prescription technology was phospholipid 44 mg，cholesterol 8 mg，paclitaxel 0.64 mg，schisandrin B 1.5 mg，ultrasonic probe time interval 5 s，prescription dosage 5 mL. According to the optimal prescription technology ，the liposomes were spherical in shape ，and the particle size was （126.49±1.19）nm，Zeta-potential was （－4.83±0.61）mV，average entrapment efficiency of liposomes was （93.88±1.67）％. Compared with RPV modified blank liposomes ，after treated with paclitaxel and schisandrin B liposomes and RPV modified paclitaxel and schisandrin B liposomes ，the survival rate ，migration inhibition rate and invasion rate of SK-OV- 3 cells were significantly decreased （P＜0.05）. The effects of RPV modified paclitaxel and schisandrin B liposomes was better than those of paclitaxel and schisandrin B liposomes （P＜0.05）. CONCLUSIONS ：RPV modified paclitaxel and schisandra B liposome are successfu lly prepared ，and they have certain antitumor activity in vitro .

Preparation and evaluation of schisandrin B-loaded F127 modified lipid-polymer nanoparticles for inhibition of breast cancer lung metastasis / 中国中药杂志

In the current study, schisandrin B(SchB)-loaded F127 modified lipid-polymer hybrid nanoparticles(SchB-F-LPNs) were developed to improve the inhibition of breast cancer lung metastasis. Modified nanoprecipitation method was used to prepare SchB-F-LPNs. The nanoparticles were spherical in shape with shell-core structure by TEM observation. SchB-F-LPNs showed a mean particle size of(234.60±6.11) nm with zeta potential of(-5.88±0.49) mV. XRD results indicated that SchB existed in the nanoparticles in an amorphous state. The apparent permeability coefficient through porcine mucus of F-LPNs was 1.43-fold of that of LPNs as shown in the in vitro mucus penetration study. The pharmacokinetics study showed that the C_(max) of SchB was(369.06±146.94) μg·L~(-1),(1 121.34±91.65) μg·L~(-1) and(2 951.91±360.53) μg·L~(-1) respectively in SchB suspensions group, SchB-LPNs group and SchB-F-LPNs group after oral administration in rats. With SchB suspensions as the reference formulation, the relative bioavailability of SchB-F-LPNs was 568.60%. SchB-F-LPNs inhibited the morphological change during transforming growth factor-β1(TGF-β1)-induced epithelial-mesenchymal transition. In addition, SchB-F-LPNs significantly decreased the number of metastatic pulmonary nodules in 4 T1 tumor-bearing mice, suggesting that SchB-F-LPNs may inhibit the metastasis of breast cancer. These results reveal the promising potential of SchB-F-LPNs in treatment of breast cancer lung metastasis.

Study on protective effect and mechanism of schisandrin B on streptozotocin induced-diabetic reinopathy / 中国免疫学杂志

Objective: To evaluate the protective effect and underlying mechanism of schisandrin B ( SB) on streptozotocin ( STZ) induced-diabetic reinopathy ( DR) in rats. Methods: Following STZ injection, rats received gavage of SB for 9 months subsequently. TUNEL assay was performed for apoptosis. The levels of SOD, MDA and LDH were determined by kits. To calculate the levels of inflammatory cytokines, ELISA assay was carried out. And western blot was performed for protein levels. Results: In STZ induced-DR rats, glucose level was makedly increased. SB inhibited STZ-induced apoptosis as demonstrated by increased apoptotic body. SB significantly increased the level of SOD and decreased the levels of MDA and LDH. Meanwhile, SB notably inhibited STZ induced-inflammation by decreasing IL-6, TNF-α and IL-1β. Furthermore, SB partly abolished STZ' s effects on the expressions of VEGF, Cdc42 and pp38. Conclusion: SB attenuates STZ induced-DR via Cdc42/p38 signaling pathway.

Evaluation of schisandrin B-induced mouse model of hypertriglyceridemia based on lipid metabolomics / 中国药理学通报

Aim: To evaluate the mouse model of hypertriglyceridemia (hTG) induced by schisandrin B (Sch B) using lipid metabolomics technology. Methods: Male ICR mice weighing 23 ~ 27 g were randomly divided into four groups: (1) mice fed with normal diet (ND group) (2) mice fed with ND and treated with Sch B(ND +Sch B group); (3) mice fed with high fat/fructose diet(HFFD group; fat, 10%; fructose, 10%; w/w), and (4) mice fed with HFFD and treated with Sch B (HFFD + Sch B group). Based on our previous research, Sch B at a single dose of 2 g · kg-1 was orally administered to the animals in the current study. Forty-eight hours later, serum samples were obtained from the orbital vein. Serum triglyceride (TG) and total cholesterol (TC) were analyzed by biochemical method. The metabolic fingerprint spectrum of serum in all groups were obtained and analyzed using ultra-performance liquid chromatography quadrupole-time-of-flight mass spectrometry (UPLC-Q/TOF-MS) method. The differences of metabolic spectra in every two groups were compared via the multivariate statistical methods. Results: Compared with ND group, three kinds (27 markers) of differential metabolites were identified in ND +Sch B group, including 18 TG, 7 phosphatidylcholine (PC), and 2 phosphatidylethano-lamine(PE). Compared with ND group, five kinds(27 markers) of differential metabolites were screened in HFFD group, including 6 sphingomyelin (SM), 13 PC, 2 cholesteryl ester(CE), 5 TG and 1 phosphati-dylinositol (PI). Compared with HFFD group, four kinds (25 markers) of differential metabolites were found in HFFD + Sch B group, involving 14 TG, 1 CE, 6 PC and 4 PE. Conclusion: These findings suggest that the animal model of hypertriglyceridemia established by Sch B involves the alteration of serum lipid metabolomics.

Antianxiety effect of schisandrin B in ICR mice and its mechanism / 吉林大学学报(医学版)

Objective: To investigate the antianxiety effect of schisandrin B (SchB) in the ICR mice, and to elucidate its related mechanisms. Methods: The ICR mice were divided into blank control group (administered with distilled water intragastrically), 2. 5 mg · kg-1 SchB group, 5 mg · kg-1 SchB group, and 10 · mg · kg-1 SchB group (administered with SchB intragastrically). After the successive administration for 7 d, cross maze test and zero maze test were conducted in all the mice, and then the peripheral blood and brain tissue samples of the mice in blank control group and 10 mg · kg-1 SchB group were taken. The levels of γ-aminobutyric acid (GABA) and glutamic acid (Glu) in the peripheral blood and brain tissue of the mice were detected by ELISA, and the ratio of GABA/Glu was calculated. The expression levels of the αl subunit of the γ-aminobutyric acid (GABAARαl) and the γ2 subunit of the γ-aminobutyric acid (GABAARγ2) in the brain tissue of the mice were detected by Western blotting method. Results: In the elevated plus maze test, the percentages of number of open arm entries in 5. 0 and 10.0 mg · kg-1 SchB groups were significantly increased compared with blank control group (P<0. 05 or P = 0. 01), and the percentages of time of open arm entries were significantly increased (P=0. 05 or P<0. 01). In the elevated zero maze test, the percentages of number of open arm entries of the mice in 5. 0 and 10. 0 mg · kg-1 SchB groups were significantly increased compared with blank control group (P<0. 01), and the percentages of number of entering open arm probe were significantly increased (P<0. 01). Compared with blank control group, the GABA levels in the peripheral blood and brain tissue of the mice in 10. 0 mg · kg-1 SchB group, were significantly increased (P<0. 01), the Glu levels in the peripheral blood and brain tissue were significantly reduced (P< 0. 01), the ratios of GABA/Glu were significantly increased (P<0. 01), and the expression levels of GABAARαl and GABAARγ2 in the brain tissue were significantly increased (P<0. 01). Conclusion: SchB has an antianxiety effect in the mice, and the effect may be related to its regulating the levels of GABA and Glu in the peripheral blood and brain tissue and the expression levels of GABAA Rα1 and GABAA Rγ2 in the brain tissue.

Effect of Schisandrin B on Proliferation and Migration of Human SW620 Colon Cancer Cell Via VEGF/PI3K/Akt Signaling Pathway / 中国药学杂志

OBJECTIVE: To study the changes of VEGF signaling pathway in colon cancer patients and the effect of schisandrin B on SW620 cells and to analyze its possible mechanism. METHODS: The protein expression of VEGFA, VEGF-R2, PI3K, Akt and p-Akt in human cancerous colon samples and adjacent normal samples were detected by Western blotting. The proliferation of SW620 cells was detected by CCK-8 method. The mRNA expression of VEGFA, VEGF-R2, PI3K and Akt in SW620 cells were detected by real-time PCR. The protein expression of VEGFA, VEGF-R2, PI3K, Akt and p-Akt in SW620 cells were detected by Western blotting. RESULTS: The protein expression of VEGF-R2, PI3K, Akt and p-Akt in human cancerous colon samples was significantly higher than that in the adjacent normal samples(P<0.05). Compared with the control group, schisandrin B could significantly inhibit the proliferation and migration of SW620 cells, the mRNA expression of VEGFA, VEGF-R2, PI3K and Akt(P<0.01) and the protein expression of VEGFA, VEGF-R2, PI3K, Akt and p-Akt(P<0.05) in SW620 cells also were significantly decreased by schisandrin B. CONCLUSION: The VEGF/PI3K/Akt signaling pathway is activated in colon cancer patients. Schisandrin B could inhibit the activity and migration of SW620 cells and inhibit the VEGF/PI3K/Akt signaling pathway.

Effects of schisandrin B on pharmacokinetics of doxorubicin in rats / 中国药科大学学报

@#To investigate the effect of combination of schisandrin B and doxorubicin on the pharmacokinetic behavior of doxorubicin in SD rats. An LC-MS/MS method was established for the determination of doxorubicin and its main metabolite doxorubicinol in SD rats plasma. Separation was performed on Agilent Eclipse XDB-C18 column(100 mm×2. 1 mm, 3. 5 μm)using 0. 1% formic acid solution and acetonitrile as mobile phase with a liner gradient program. The ion transitions were performed under ESI position model at m/z 544. 2→397. 3(doxorubicin), m/z 546. 2→399. 2(doxorubicinol), m/z 528. 2→381. 2(daunorubicin, internal standard). Calibration curves of doxorubicin(0. 500-500 ng/mL)and doxorubicinol(0. 200-50. 0 ng/mL)showed good linear regression. The precision and accuracy met the requirements. The variation coefficient of extraction recovery and matrix effect was less than 15%. The AUC0-t of doxorubicin were(605. 69±145. 84)and(564. 53±23. 99)ng ·h/mL in alone and combined group, respectively. The AUC0-t of doxorubicinol were(26. 69±13. 41)and(29. 00±2. 78)ng ·h/mL, respectively. Results indicated that, schisandrin B had not affected the pharmacokinetic behavior of doxorubicin in SD rats.

Anti-inflammatory and Immune Function Enhancing Effects of Schisandrin B in Mice / 中国药师

Objective: To assess the anti-inflammatory and immunomodulatory activities of schisandrin B in mice. Methods: Kun-ming mice were randomly divided into 6 groups: the blank control group, the model control group, the positive control group( dexam-ethasone hydrochloride 5 mg·kg-1,levamisole hydrochloride 30 mg·kg-1), schisandrin B low (100 mg·kg-1), medium (200 mg ·kg-1) and high (400 mg·kg-1) dose groups. The inflammatory mice were induced by xylene and the immunosuppressed mice were induced by cyclophosphamide. The ear edema experiment, carbon clearance experiment and serum hemolysin experiment were per-formed with the swelling rate, swelling inhibition rate, carbon clearance index, phagocytic index, half hemolytic value, spleen index and thymus index as the investigation factors. Results: High dose and medium dose schisandrin B groups had significant inhibitory effect on mice ear edema when compared with the model control group(P<0. 05);There were significant differences between the high dose group and the positive group and there were significant differences in clearance index and phayxyitc index between high dose group and low dose group(P<0. 05), and high dose schisandrin B group had increased clearance index and phagocytic index in low immune function mice when compared with the model control group (P<0.05). Schisandrin B significantly increased HC50in low immune function mice when compared with the model control group (P<0. 01). Schisandrin B significantly increased the spleen index and thy-mus index in low immune function mice when compared with the model control group ( P<0. 01,and in a dose-dependent manner. Conclusion: Schisandrin B has significant anti-inflammatory and immune function enhancing effects.

Antianxiety effect of schisandrin B in ICR mice and its mechanism / 吉林大学学报(医学版)

Objective:To investigate the antianxiety effect of schisandrin B (SchB) in the ICR mice,and to elucidate its related mechanisms.Methods:The ICR mice were divided into blank control group (administered with distilled water intragastrically),2.5 mg · kg-1 SchB group,5 mg · kg-1 SchB group,and 10 · mg · kg-1 SchB group (administered with SchB intragastrically).After the successive administration for 7 d,cross maze test and zero maze test were conducted in all the mice,and then the peripheral blood and brain tissue samples of the mice in blank control group and 10 mg · kg-1 SchB group were taken.The levels of γ-aminobutyric acid (GABA) and glutamic acid (Glu) in the peripheral blood and brain tissue of the mice were detected by ELISA,and the ratio of GABA/Glu was calculated.The expression levels of the α1 subunit of the γ-aminobutyric acid (GABAARα1) and the γ2 subunit of the γ-aminobutyric acid (GABAARγ2) in the brain tissue of the mice were detected by Western blotting method.Results:In the elevated plus maze test,the percentages of number of open arm entries in 5.0 and 10.0 mg · kg 1 SchB groups were significantly increased compared with blank control group (P＜0.05 or P=0.01),and the percentages of time of open arm entries were significantly increased (P=0.05 or P＜0.01).In the elevated zero maze test,the percentages of number of open arm entries of the mice in 5.0 and 10.0 mg · kg-1SchB groups were significantly increased compared with blank control group (P＜0.01),and the percentages of number of entering open arm probe were significantly increased (P＜0.01).Compared with blank control group,the GABA levels in the peripheral blood and brain tissue of the mice in 10.0 mg · kg-1 SchB group,were significantly increased (P＜0.01),the Glu levels in the peripheral blood and brain tissue were significantly reduced (P＜0.01),the ratios of GABA/Glu were significantly increased (P＜0.01),and the expression levels of GABAARα1and GABAARγ2 in the brain tissue were significantly increased (P＜0.01).Conclusion:SchB has an antianxiety effect in the mice,and the effect may be related to its regulating the levels of GABA and Glu in the peripheral blood and brain tissue and the expression levels of GABAA Rα1 and GABAA Rγ2 in the brain tissue.

Beneficial effects of Schisandrin B on the cardiac structure and function in a mice model of myocardial infarction / 中华心血管病杂志

Objective@#To investigate whether Schisandrin B (Sch B) could improve cardiac structure and function in myocardial infarction (MI) mice and related mechanisms.@*Methods@#Male C57BL/6J mice were randomized into sham (n=8), MI+ Sch B (n=24, 80 mg·kg-1·d-1 per gavage) or MI+ vehicle (n=24, equal volume). After treatment for 3 weeks, cardiac function was detected by echocardiography measurement.Infarction size was measured by Evans blue and TTC staining.HE and Masson trichrome staining were used to observe the myocardial inflammation, structure and fibrosis.TNF-α, TGF-β, IL-1β were detected by ELISA. The apoptosis index of ischemic myocardial cells was detected by immunofluorescence. NF-κB, Bcl-2, Bax, Akt phosphorylated Akt(p-Akt), eNOS, phosphorylated eNOS (p-eNOS) were detected by Western blot.@*Results@#Three weeks after operation, survival rate (83.33% vs. 54.17%, P<0.05), LVEF((51.77±8.50)% vs.(40.23±8.30)%, P<0.05), LVFS((26.44±5.15)% vs. (19.53±4.56)%, P<0.05)were significantly higher; LVEDD ((4.13±0.40) mm vs.(4.44±0.46)mm, P<0.05), LVESD((3.07±0.39) mm vs. (3.46±0.52)mm, P<0.05), the heart weight/body weight ratio((0.59±0.06)% vs. (0.68±0.10)%, P<0.05)was significantly lower and infarct size ((23.4±2.36)% vs. (39.4±2.06)%, P<0.05) was significantly reduced in MI+ Sch B group than those in MI+ vehicle group. In MI+ vehicle group, HE staining showed a large number of inflammatory cells in the peri-infarctl region, and the permutation structure was very disorganized, while above changes were significantly reduced in the MI+ Sch B group. Masson staining results showed that the degree of myocardial fibrosis in MI+ Sch B group was significantly less than that of MI+ vehicle group.Moreover, Sch B could down-regulate some inflammatory cytokines, like NF-κB、TGF-β、TNF-α and IL-1β, activate Akt-eNOS pathway, upgrade Bcl-2 and downgrade Bax and reduce cell apoptosis as compared with MI+ vehicle group (all P<0.05).@*Conclusions@#Our results demonstrate that Sch B can reduce inflammation, inhibit apoptosis, and attenuate cardiac remodeling and improve cardiac function in this mice MI model.Sch B might serve as a potential novel therapeutic agent for ischemic heart disease.

Co-Delivery of Doxorubicin and Schisandrin B by Liposome to Overcome Multidrug Resistance / 中国药学杂志

OBJECTIVE: To investigate the synergetic effect of schisandrin B and liposomes on overcoming multidrug resistance(MDR). METHODS: Co-delivery of doxorubicin and schisandrin B by liposome(DS-L) were prepared, and doxorubicin solution(F-DOX), schisandrin B and doxorubicin mixture(Sch B+DOX), doxorubicin liposomes(D-L), doxorubicin liposomes and schisandrin B mixture(D-L+Sch B), verapamil and doxorubicin mixture(Ver+DOX) were also prepared as the control. The MTT test were measured, and the amount of doxorubicin in the K562/DOX cells at different time were determined, and time course of uptake and efflux were drawn. RESULTS: The MTT test shows that the resistance factor(RF) of group DS-L were 1.68, 14.52 and 1.42 times of group Sch B+DOX, group D-L and group D-L+Sch B respectively. The uptake test shows that amount of doxorubicin of K562/DOX cells in group DS-L was 1.30 and 1.21 times of that in group D-L and group Sch B+DOX respectively. And the efflux test shows DS-L could delay doxorubicin efflux from K562/DOX cells. CONCLUSION: The co-delivery of chemotherapeutics and P-gp inhibitors schisandrin B by liposome is a promising approach to overcome MDR. And there is a synergistic effect between liposome and schisandrin B to overcome MDR.

Effects of schisandrin B on P21 and Caspase-3 expression in HK-2 cells injuried by CDDP / 中草药

Objective: To explore the effect of schisandrin B (SchB) on P21 and Caspase-3 expression in cisplatin induced human proximal renal tubular epithelial (HK-2) cells injuried by CDDP in vitro. Methods: HK-2 cells were randomly divided into five groups: control group (untreated), CDDP group (treated with 50 μmol/L CDDP for 24 h), SchB pretreat groups (CDDP + SchB, cells were pretreated with 5, 10, and 20 μmol/L SchB for 2 h, and the rest of the operation was the same as CDDP group). CCK-8 kit was used to detect the cell viability of five groups. Flow cytometry was used to detect the apoptotic rate of five groups. The morphology of cells was observed by inverted microscope. The protein expression of P21 and Caspase-3 was assessed by Western blotting assay. Results: Compared with the control group, the cell viability decreased, and the apoptosis increased, the protein expression of P21 and Caspase-3 was up-regulated in HK-2 cells after treated with 50 μmol/L CDDP. Cell volume was reduced, and the mutual connection disappeared. Compared with the CDDP group, the cell viability increased and the apoptosis was decreased, the expression of Caspase-3 was down-regulated and the expression of P21 was up-regulated in HK-2 cells after pretreated with 5, 10, and 20 μmol/L SchB. Cell morphological damage lightened, the volume slightly decreased, and the mutual connection reduced. Conclusion: SchB has a protective effect on HK-2 cells damage induced by cisplatin. The mechanism may be related to up-regulation of P21 and down-regulation of Caspase-3 expression, and its protective effect is dose dependent.

Anti-oxidative damage effect in Schisandrin B in mice of Alzheimer's disease and its mechanism / 吉林大学学报(医学版)

Objective:To investigate the preventive and protective effects of Schisandrin B in the mice of Alzheimer's disease (AD),and to clarify its mechanism.Methods:Fifty Balb/c mice were randomly divided into blank group,model group,pasitive control group,low dose of Schisandrin B group(0.1 g·kg-1)and high dose of Schisandrin B group(0.5 g·kg-1);there were 10 mice in each group.Step-through test was conducted after last administration to detect the latencies and number of errors of the mice in various groups,and the brain tissue was taken.Congo red staining was to detect the morphology changes of cells and neuronal amyloidosis in brain tissue of the mice.The levels of ROS in brain tissue of the mice were tested by Flow Cytometry.The contents of MDA,the levels of LDH,and the activities of CAT,GSH-Px and SOD in brain tissue of the mice were tested by biochemical method.Western blotting method was used to detect the expression levels of signaling pathway proteins Nrf2 and Keap1 in brain tissue of the mice.Results:Compared with model group,the latencies of the mice in low and high dose of Schisandrin B groups were increased (P<0.01) and the number of errors in step-through tests was decreased (P<0.05 or P<0.01).The Congo red staining results showed that compared with model group,the neuronal amyloidosis in brain tissue of the mice in Schisandrin B groups was decreased significantly.Compared with model group,the levels of ROS,LDH and the contents of MDA in brain tissue of the mice in low and high doses of Schisandrin B groups were decreased (P<0.05 or P<0.01),and the activities of CAT,SOD and GSH-Px were increased (P<0.01).Compared with low dose of Schisandrin B group,the content of MDA and the activities of SOD and GSH-Px in brain tissue of the mice in high dose of Schisandrin B group were increased (P<0.001).Compared with model group,the expression level of Nrf2 protein in brain tissue of the mice in low dose of Schisandrin B group was increased (P<0.01),while the expression level of Nrf2 protein in brain tissue of the mice in high dose of Schisandrin B group was decreased (P<0.01);the expression levels of Keap1 protein in brain tissue of the mice in low and high doses of Schisandrin B groups was decreased (P<0.01).Conclusion:Schisandrin B could decrease the level of peroxidation in brain tissue of the mice and reduce the oxidative damage and improve the memory function of the AD mice.The mechanism is related to the activation of Nrf2 signaling pathway which improve the activity of antioxidant enzymes.

Protective effect of schisandrin B on cerebral ischemia reperfusion injury of rats and its mechanisms / 吉林大学学报(医学版)

Objective:To investigate the protective effect of schisandrin B (SchB)on the cerebral ischemia reperfusion injury of the rats and the influence in HSPA12B/PI3K/Akt signaling pathway,and to explore the mechanisms.Methods:130 SD rats were divided into sham group,cerebral ischemia reperfusion injury model group (model group),low dose of SchB group (SchB 3 mg· kg-1 ,SchB1 group),middle dose of SchB group (SchB 10 mg·kg-1 ,SchB2 group)and high dose of SchB group (SchB 30 mg·kg-1 ,SchB3 group)(n=26).The rats in sham group didn’t plug lines;the rats in model were used to establish ischemia reperfusion models;the rats in SchB1 ,SchB2 and SchB3 groups were firstly pretreated with different doses of SchB for 7 d,and then they were used to build cerebral ischemia reperfusion injury models.The nerve dysfunction of rats was evaluated by neurologic deficit score.The cerebral edema was detected by measuring the content of water in brain tissue.The morphological changes of brain tissue were observed by toluidine blue staining.The levels of nuclear factor kappa B (NF-κB), tumor necrosis factor-α(TNF-α),interleukin-1 (IL-1)and interleukin-6 (IL-6)in the brain tissue were detected by ELISA.Western blotting method was used to detect the protein expression levels of heat shock protein A12B (HSPA12B ), serine-threonine kinase (Akt ) and phosphorylation serine-threonine kinase (p-AKT ). Results:Compared with sham group,the neurologic deficit score of rats in model group was significantly increased (P <0.01),and the content of water in brain tissue was increased (P < 0.01 );the brain tissue structure was loosened,and the mesenchyme appeared edema;the NF-κB,TNF-α,IL-1,and IL-6 levels were increased (P <0.01),and the expression levels of HSPA12B and p-Akt proteins were decreased (P <0.01).Compared with model group,the neurologic deficit scores of the rats in SchB1 ,SchB2 ,and SchB3 groups were decreased (P <0.01),and the contents of water in brain tissue of the rats in SchB2 and SchB3 groups were decreased (P <0.05);the edema of nerve cells was alleviated,and the cavities were reduced;the NF-κB,TNF-α,IL-1,and IL-6 levels were decreased (P <0.05 or P <0.01),the expression levels of the HSPA12B protein in SchB2 ,and SchB3 groups were increased (P <0.05),and the p-Akt protein expression levels of the rats in SchB1 ,SchB2 ,and SchB3 groups were increased (P <0.01).Conclusion:SchB could protect the cerebral ischemia reperfusion injury of rats,its mechanism may be related to regulating the HSPA12B/PI3K/Akt signaling pathway and inhibiting the inflammatory reaction damage to the nerve cells of reperfusion.

Simultaneous Determination of Five Effective Components in Hugan Tablets by HPLC / 中国药师

Objective:To establish an HPLC method for the simultaneous determination of adenosine, schisandrin, schisantherin,deoxyschizandrin and schisandrin B in Hugan tablets. Methods:An Agilent Eclipse XDB column(250 mm × 4. 6 mm,5 μm)was used with the mobile phase of acetonitrile- 0. 7% phosphoric acid solution with gradient elution. The flow rate was 0. 80 ml·min -1 . The detection wavelength was set at 260 nm in 0-14 min and 250 nm in 14-58 min,the column temperature was 30℃ ,and the sample size was 10 μl. Results:There was a good linear relationship within the range of 2. 35- 47. 00 μg· ml -1(r = 0. 999 8)for adenosine,14. 90-297. 00 μg·ml -1(r = 1. 000 0)for schisandrin,3. 46- 69. 20 μg·ml-1(r = 0. 999 9)for schisantherin,4. 00- 80. 10 μg·ml -1(r = 1. 000 0)for deoxyschizandrin and 3. 42- 68. 30 μg·ml-1(r = 0. 999 9)for schisandrin B. The average recoveries for the five components were all above 98% . Conclusion:The method is simple and accurate with good reproducibility,which can be used for the determination of adenosine,schisandrin,schisantherin,deoxyschizandrin and schisandrin B in Hugan tablets.