Systems pharmacology-based investigation of Sanwei Ganjiang Prescription: related mechanisms in liver injury.

Liver injury remains a significant global health problem and has a variety of causes, including oxidative stress (OS), inflammation, and apoptosis of liver cells. There is currently no curative therapy for this disorder. Sanwei Ganjiang Prescription (SWGJP), derived from traditional Chinese medicine (TCM), has shown its effectiveness in long-term liver damage therapy, although the underlying molecular mechanisms are still not fully understood. To explore the underlining mechanisms of action for SWGJP in liver injury from a holistic view, in the present study, a systems pharmacology approach was developed, which involved drug target identification and multilevel data integration analysis. Using a comprehensive systems approach, we identified 43 candidate compounds in SWGJP and 408 corresponding potential targets. We further deciphered the mechanisms of SWGJP in treating liver injury, including compound-target network analysis, target-function network analysis, and integrated pathways analysis. We deduced that SWGJP may protect hepatocytes through several functional modules involved in liver injury integrated-pathway, such as Nrf2-dependent anti-oxidative stress module. Notably, systems pharmacology provides an alternative way to investigate the complex action mode of TCM.

Implementation of an interprofessional team-based learning program involving seven undergraduate health and social care programs from two universities, and students' evaluation of their readiness for interprofessional learning.

BACKGROUND: Interprofessional learning is gaining momentum in revolutionizing healthcare education. During the academic year 2015/16, seven undergraduate-entry health and social care programs from two universities in Hong Kong took part in an interprofessional education program. Based on considerations such as the large number of students involved and the need to incorporate adult learning principles, team-based learning was adopted as the pedagogy for the program, which was therefore called the interprofessional team-based learning program (IPTBL). The authors describe the development and implementation of the IPTBL program and evaluate the effectiveness of the program implementation. METHODS: Eight hundred and one students, who are predominantly Chinese, participated in the IPTBL. The quantitative design (a pretest-posttest experimental design) was utilized to examine the students' gains on their readiness to engage in interprofessional education (IPE). RESULTS: Three instructional units (IUs) were implemented, each around a clinical area which could engage students from complementary health and social care disciplines. Each IU followed a team-based learning (TBL) process: pre-class study, individual readiness assurance test, team readiness assurance test, appeal, feedback, and application exercise. An electronic platform was developed and was progressively introduced in the three IUs. The students' self-perceived attainment of the IPE learning outcomes was high. Across all four subscales of RIPLS, there was significant improvement in student's readiness to engage in interprofessional learning after the IPTBL. A number of challenges were identified: significant time involvement of the teachers, difficulty in matching students from different programs, difficulty in making IPTBL count towards a summative assessment score, difficulty in developing the LAMS platform, logistics difficulty in managing paper TBL, and inappropriateness of the venue. CONCLUSIONS: Despite some challenges in developing and implementing the IPTBL program, our experience showed that TBL is a viable pedagogy to be used in interprofessional education involving hundreds of students. The significant improvement in all four subscales of RIPLS showed the effects of the IPTBL program in preparing students for collaborative practice. Factors that contributed to the success of the use of TBL for IPE are discussed.

One-Compound-Multi-Target: Combination Prospect of Natural Compounds with Thrombolytic Therapy in Acute Ischemic Stroke.

Tissue plasminogen activator (t-PA) is the only FDA-approved drug for acute ischemic stroke treatment, but its clinical use is limited due to the narrow therapeutic time window and severe adverse effects, including hemorrhagic transformation (HT) and neurotoxicity. One of the potential resolutions is to use adjunct therapies to reduce the side effects and extend t-PA's therapeutic time window. However, therapies modulating single target seem not to be satisfied, and a multitarget strategy is warranted to resolve such complex disease. Recently, large amount of efforts have been made to explore the active compounds from herbal supplements to treat ischemic stroke. Some natural compounds revealed both neuro- and bloodbrain- barrier (BBB)-protective effects by concurrently targeting multiple cellular signaling pathways in cerebral ischemia-reperfusion injury. Thus, those compounds are potential to be one-drug-multi-target agents as combined therapy with t-PA for ischemic stroke. In this review article, we summarize current progress about molecular targets involving in t-PA-mediated HT and neurotoxicity in ischemic brain injury. Based on these targets, we select 23 promising compounds from currently available literature with the bioactivities simultaneously targeting several important molecular targets. We propose that those compounds merit further investigation as combined therapy with t-PA. Finally, we discuss the potential drawbacks of the natural compounds' studies and raise several important issues to be addressed in the future for the development of natural compound as an adjunct therapy.

High expression of erythropoietin-producing hepatoma cell line-B2 (EphB2) predicts the efficiency of the Qingyihuaji formula treatment in pancreatic cancer CFPAC-1 cells through the EphrinB1-EphB2 pathway.

Our previous study demonstrated that inhibition of erythropoietin-producing hepatoma cell line-B2 (EphB2) expression resulted in the promotion of cancer growth, with EphB2 acting as a tumor suppressor in pancreatic cancer. Qingyihuaji formula (QYHJ), a traditional Chinese medicine, acts as an independent protective factor for pancreatic cancer patient survival and different patients have shown various responses to QYHJ treatment. In the current study, the different effects on tumor growth inhibition following QYHJ treatment in cells with different levels of EphB2 expression were investigated to reveal the mechanism. A subcutaneously transplanted tumor model using cancer cells with different levels of EphB2 expression were established in vivo and received a four-week QYHJ intervention. Tumor weight inhibitory rate and tumor volume deflation were evaluated. The cell cycle and apoptosis were analyzed by flow cytometry, and reverse transcription polymerase chain reaction and western blot analysis were used to assess mRNA and protein levels. The results showed that the tumor weight inhibitory rate was 31.40, 31.33 and 18.36% in CFPAC-1, CFPAC-1 control RNAi and CFPAC-1 EphB2 RNAi cells following QYHJ treatment, respectively. A statistically significant difference was identified in CFPAC-1 (P<0.05) and CFPAC-1 control RNAi (P<0.01) cells. In addition, a statistically significant increase was identified in the G0/G1 phase population (P<0.05) and a statistically significant decrease was identified in the S phase population (P<0.05) in CFPAC-1 and CFPAC-1 control RNAi cells; however, no significant difference was identified in the CFPAC-1 EphB2 RNAi cells following QYHJ treatment. QYHJ upregulated the mRNA and protein level of Eph receptor-interacting B1 (EphrinB1) in the cells that were expressing different levels of EphB2, however, QYHJ did not regulate EphB2 expression. In CFPAC-1 and CFPAC-1 control RNAi cells, the QYHJ treatment resulted in a statistically significant decrease in cyclin-dependent kinase 6 (CDK6) mRNA (P<0.05) and protein (P<0.05) levels. The high expression of EphB2 predicted the superior response rate to the QYHJ treatment through a mechanism of inhibiting the cell cycle by an EphrinB1-EphB2-induced CDK6 decrease in CFPAC-1 cells. Therefore, EphB2 acts as a predictive factor for QYHJ treatment in pancreatic cancer CFPAC-1 cells.

Anti-inflammatory effects of naringin in chronic pulmonary neutrophilic inflammation in cigarette smoke-exposed rats.

Naringin, a well-known flavanone glycoside of grapefruit and citrus fruits, was found to be as an effective anti-inflammatory compound in our previous lipopolysaccharide-induced acute lung injury mouse model via blockading activity of nuclear factor κB. The current study sought to explore the anti-inflammatory effects of naringin on chronic pulmonary neutrophilic inflammation in cigarette smoke (CS)-induced rats. Seventy Sprague-Dawley rats were randomly divided into seven groups to study the effects of CS with or without various concentrations of naringin or saline for 8 weeks. The results revealed that naringin supplementation at 20, 40, and 80 mg/kg significantly increased body weight of CS-induced rats as compared to that in the CS group. Moreover, naringin of 20, 40, and 80 mg/kg prevented CS-induced infiltration of neutrophils and activation of myeloperoxidase and matrix metalloproteinase-9, in parallel with suppression of the release of cytokines, such as tumor necrosis factor-α and interleukin-8 (IL-8). IL-10 in bronchoalveolar lavage fluid was significantly suppressed after CS exposure, but dose dependently elevated by naringin. The results from hematoxylin and eosin staining revealed that naringin dose dependently reduced CS-induced infiltration of inflammatory cells, thickening of the bronchial wall, and expansion of average alveolar airspace. In conclusion, our data suggest that naringin is an effective anti-inflammatory compound for attenuating chronic pulmonary neutrophilic inflammation in CS-induced rats.

Spatholobus suberectus inhibits cancer cell growth by inducing apoptosis and arresting cell cycle at G2/M checkpoint.

AIM OF THE STUDY: Although herbs have long been alternatively applied for cancer treatment in China, its treatment effects and their potential mechanisms have not been sufficiently investigated. The chinese herb Spatholobus suberectus (SS) is commonly prescribed to cancer patients. In this study, the anti-cancer effect of SS and its molecular mechanisms have been investigated. MATERIALS AND METHODS: The effect of SS on cell proliferation was studied by cell growth assay and flow cytometry on breast cancer cell lines MCF-7 and colon cancer cell line HT-29. The role of SS in apoptosis was studied by flow cytometry, DNA fragmentation assay and mitochondrial membrane potential assay. Expression of proteins associated with cell cycle and apoptosis was determined by Western blot analysis. The in vivo effect of SS was tested in nude mouse cancer xenografts. RESULTS: Cell growth assay showed that SS effectively inhibits tumor cell growth in a dose-dependent manner. Flow cytometry analysis showed that SS could arrest the cell cycle at G2/M checkpoint, which is associated with DNA damage and activation of phosphor-Chk1/Chk2. The pro-apoptotic effect of SS was demonstrated by Annexin V-PI staining and mitochondrial membrane potential assay. In vivo experiments show that the efficiency of SS alone group was superior to docetaxel or to docetaxel and SS combined. No obvious body weight loss or blood toxicity was observed in SS tested animals. CONCLUSIONS: Our data demonstrates that SS is a potential herb for cancer treatment by inhibiting tumor growth via induction of apoptosis and arrest of the cell cycle at G2/M phase.

Different mechanisms of cis-9,trans-11- and trans-10,cis-12- conjugated linoleic acid affecting lipid metabolism in 3T3-L1 cells.

Conjugated linoleic acid (CLA) has been shown to reduce body fat mass in various experimental animals. It is valuable to identify its influence on enzymes involved in energy expenditure, apoptosis, fatty acid oxidation and lipolysis. We investigated isomer-specific effects of high dose, long treatment of CLA (75.4 µmol/L, 8 days) on protein and gene expression of these enzymes in cultured 3T3-L1 cells. Proteomics identified significant up- or down-regulation of 52 proteins by either CLA isomer. Protein and gene expression of uncoupling protein (UCP) 1, UCP3, perilipin and peroxisome proliferator-activated receptor (PPAR) α increased whereas UCP2 reduced for both CLA isomers. And eight-day treatment of trans-10,cis-12 CLA, but not cis-9,trans-11 CLA, significantly up-regulated protein and mRNA levels of PKA (P<.05), CPT-1 and TNF-α (P<.01). Compared to protein expression, both isomers did not significantly influence the mRNA expression of HSL, ATGL, ACO and leptin. In conclusion, high-dose, long treatment of cis-9,trans-11 CLA did not promote apoptosis, fatty acid oxidation and lipolysis in adipocytes, but may induce an increase in energy expenditure. trans-10,cis-12 CLA exhibited greater influence on lipid metabolism, stimulated adipocyte energy expenditure, apoptosis and fatty acid oxidation, but its effect on lipolysis was not obvious.

Characterization of chemical components in extracts from Si-wu decoction with proliferation-promoting effects on rat mesenchymal stem cells.

Si-wu decoction (SDE), a classic prescription in Traditional Chinese Medicine, has been used for the treatment of a variety of anaemia in China for centuries. In order to explore the scientific basis of the formula, we investigated the relationship between its chemical components and proliferation-promoting effects on rat marrow-derived mesenchymal stem cells (MSCs). Twenty (F-1-F-20) components were obtained and their proliferation-promoting effects on MSCs were investigated. The results showed that F-4, F-7, F-10, and F-11 stimulated the proliferation of the MSCs. The chemical components with proliferation-promoting effects on the MSCs were further identified by GC-MS, HPLC, LC-MS, and other spectra. Ligustilide (F-4) isolated from SDE showed the best proliferation-promoting effect. Palmitic acid methyl ester and stearic acid ethyl identified from F-7 and F-10 by HPLC were also confirmed to be responsible for stimulating MSC proliferation. A novel compound, 6,7-dihydroxy-3-(-prop-1-enyl)isobenzofuran-1(3H)-one, was found in the SDE for the first time by LC-MS(n), whose structure was similar to ligustilide.

Effectiveness and safety of herbal medicines in the treatment of irritable bowel syndrome: a systematic review.

AIM: To explore the efficacy and safety of herbal medicines (HM) in the treatment of irritable bowel syndrome (IBS). METHODS: A computer-based as well as manual literature search was performed. We reviewed randomized controlled trials on the treatment of IBS with and without HM. RESULTS: A total of 22 studies with 25 HMs met the inclusion criteria. Four of these studies were of good quality, while the remaining 18 studies involving 17 Chinese herbal medicine (CHM) formulas were of poor quality. Eight of these reports using 9 HMs showed global improvement of IBS symptoms, 4 studies with 3 HMs were efficacious in diarrhea-predominant IBS, and 2 studies with 2 HMs showed improvement in constipation-predominant IBS. Out of a total of 1279 patients, 15 adverse events in 47 subjects were reported with HM. No serious adverse events or abnormal laboratory tests were observed. The incidence of the adverse events was low (2.97%; 95% CI: 2.04%-3.90%). CONCLUSION: Herbal medicines have therapeutic benefit in IBS, and adverse events are seldom reported in literature. Nevertheless, herbal medicines should be used with caution. It is necessary to conduct rigorous, well-designed clinical trials to evaluate their effectiveness and safety in the treatment of IBS.

[Comparative study of Buyang Huanwu Decoction and the different combinations of its ingredients on neurogenesis following ischemic stroke in rats].

OBJECTIVE: To investigate the effect of Buyang Huanwu Decoction (BYHWD) and the different combinations of its ingredients on neurogenesis following ischemic stroke in rats. METHODS: The model rats of ischemic stroke was established by blocking cerebral media artery with electrocoagulation through craniectomy, and electric stimulation, given from 24 h after blocking, 2 h daily for 15 successive days. They were divided into four groups, Group A treated with saline, Group B treated with BYHWD, Group C treated with BYHWD but earthworm subtracted, and Group D treated with Danggui Buxue Decoction (DGBXD). The expression of 5-bromodeoxyuridine (BrdU) in cerebral tissue was determined by immunohistochemical method. RESULTS: Large amount of BrdU immunoreactive cells presented in the hippocampal region of rats in Group B and C, densely arranged, partial in cluster, with the figure significantly different to that in Group A (P < 0.01), and the amount in the ischemic side was significantly more than that in the opposite side (P < 0.05). While comparing between Group A and D, the amount of BrdU immunoreactive cells in the hippocampal region showed insignificant difference (P > 0.05). CONCLUSION: BYHWD has a effect in promoting neurogenesis better than DGBXD.

[Effects of Buyanghuanwu decoction on nerve proliferation in rats with sequelae of ischemic stroke].

OBJECTIVE: To examine the effect of Buyanghuanwu decoction (BYHWD) in inducing nerve proliferation in rats with sequelae of ischemic stroke. METHODS: A rat model of ischemic stroke sequelae was established by means of craniectomy in which the right common carotid artery was ligated with 4-0 silk thread followed by cauterization of the right middle cerebral artery. Programmed electric shock was administered 24 h after the onset of ischemic stroke for 2 h daily for 20 consecutive days. The rats in sham operation group were not subjected to ligation of the right common carotid artery or right middle cerebral artery occlusion. The rats in the treatment groups were given oral BYHWD for 15 consecutive days. All the rats received repeated intraperitoneal injections of the cell proliferation-specific marker 5-bromodeoxyuridine (BrdU), and the intake of BrdU in the cerebral tissues was determined by immunohistochemistry. RESULTS: The number of BrdU-immunoreactive cells in the cerebral tissues of BYHWD-treated rats was significantly greater than that in the untreated model group. CONCLUSION: BYHWD can promote nerve proliferation in rats with ischemic stroke sequelae.

[Effect of buyang huanwu decoction drug serum on expression of p53 and p21 genes in cultured rat's cerebral cortical neuron after hypoxia in vitro].

OBJECTIVE: To explore the effect of Buyang Huanwu decoction (BHD) drug serum on rat's in vitro cultured cerebral cortical neuron apoptosis induced by hypoxia, and on the expression of p53 and p21 genes in hypoxia process. METHODS: The model of hypoxia neuron apoptosis was established adopting Daniel method and treated with BHD drug serum. The neuron apoptosis rate was determined by flow cytometry with propidium iodide staining, the p53 and p21 gene expression was tested by immunohistochemical method with flow cytometry. RESULTS: BHD could significantly inhibit the neuron apoptosis induced by hypoxia and down-regulate the expressions of p53 and p21 genes. CONCLUSION: BHD shows inhibition on neuron hypoxia apoptosis and down-regulating of the p53 and p21 gene expression is one of its mechanisms.

[Effects of Buyanghuanwu decoction (BYHWT) on proliferation of cultured rat cortical neurons].

OBJECTIVE: To observe the effect of rat serum containing Buyanghuanwu decoction (BYHWT) on the proliferation of cultured rat cortical neurons, so as to understand the mechanism of BYHWT in the treatment of hypoxia brain damage. METHODS: The growth of cultured rat cortical neurons were observed by MTT assay to evaluate the effect of the serum containing BYHWT on the neurons cultured in both normal and hypoxia conditions. RESULTS: BYHWT significantly promoted proliferation of the neurons cultured under both normal and hypoxia conditions, in comparison with the response of the cells to drug-free serum (P<0.05). CONCLUSION: Some of the constituents of BYHWT in rat serum can promote the proliferation of rat cortical neurons cultured in both normal and hypoxia conditions.

[Protective effect of Buyanghuanwu Tang on cultured rat cortical neurons against hypoxiainduced apoptosis].

OBJECTIVE: To investigate the protective effect of Buyanghuanwu Tang (BYHWT), a decoction with 6 herbal ingredients, on in vitro cultured rat cortical neurons against apoptosis induced by hypoxia, and study its effect on the production of nitric oxide (NO) and oxygen free radicals and Bcl-2 expression in the neurons during hypoxia. METHODS: Models of hypoxia-induced neuron apoptosis were established and treated with sera containing BYHWT. Stained by propidium iodide, the neurons underwent apoptosis analysis using flow cytometry, and the levels of malonyl dialdehyde (MDA) and NO were measured with spectrophotometer, with Bcl-2 expression assayed by immunohistochemical method with flow cytometry. RESULTS: In BYHWT-treated neurons, apoptosis rates were significantly lower than those of the neurons subjected to hypoxia exclusively, and at the same time NO and MDA production was remarkably reduced. Bcl-2 expression, however, was up-regulated. CONCLUSION: BYHWT can protect neurons from hypoxia-induced apoptosis, the mechanism of which may lie in the elimination of NO and oxygen free radicals produced during hypoxia and up-regulation of Bcl-2 expression.

[Study on preventive effect of buyang huanwu decoction on cardiomyocyte apoptosis induced by hypoxia-reoxygenation in rats].

OBJECTIVE: To observe the preventive effect of rats' serum containing Buyang Huanwu Decoction (BYHWD) on cultured cardiomyocyte apoptosis of neonatal rat induced by means of 24 hrs hypoxia and 4 hrs reoxygenation, and to investigate its mechanism concerned with nitric oxide (NO). METHODS: Myocyte apoptosis was detected by flow cytometry and ELISA with Annexin V-PI double labeled method. The lactate dehydrogenase (LDH) releasing level was measured with ultraviolet spectrophotometer. The NO concentration was determined by modified Yu method and the concentration of thiobarbituric acid response substance (TBARS) was tested by Ohkawa method. RESULTS: BYHWD contained rats' serum could significantly prevent cardiomyocyte from apoptosis induced by hypoxia and reoxygenation. After hypoxia-reoxygenation, the NO, LDH and TBARS levels in the supernatant of cultured liquid treated with BYHWD were significantly lower than those in non-treated cultured liquid, the effect of BYHWD was dose-dependent. CONCLUSION: BYHWD can prevent cardiomyocytes from apoptosis induced by hypoxia and reoxygenation, its mechanism might be related with oxygen free radical and NO scavenging produced during the hypoxia-reoxygenation process.