Systems pharmacology-based investigation of Sanwei Ganjiang Prescription: related mechanisms in liver injury / 中国天然药物

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.

Systems pharmacology-based investigation of Sanwei Ganjiang Prescription: related mechanisms in liver injury / 中国天然药物

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.

FAK-related non-kinase plasmid transfection inhibited hepatic stellate cells proliferation / 中国应用生理学杂志

<p><b>AIM</b>To observe the effect of FAK-related non-Kinase (FRNK) plasmid on hepatic stellate cell (HSC) proliferation stimulated by fibronectin (FN).</p><p><b>METHODS</b>FRNK plasmid was transfected into HSC with transient liposomal transfection. The proteins of FRNK, FAK and p-FAK(Tyr397) were assayed by Western blotting analysis. The proliferation of HSC was evaluated by improved MTT assay, and cell cycle pattern was determined by flow cytometry (FCM).</p><p><b>RESULTS</b>(1) The expression of FRNK protein increased after FRNK transfected HSC, and it was at 48 h that the expression of FRNK protein was the highest (P < 0.01). The protein level of FAK was no significant difference between before FRNK plasmid transfection and after transfection (P > 0.05). The expression of p-FAK(Tyr397) protein was down-regulated after FRNK had been transfected in HSC, (P < 0.01). (2) The HSC proliferation inhibition rates at 12 h, 24 h and 48 h after FRNK transfection were 20.07%, 26.16%, 29.77%, respectively (P < 0.01). (3) Compared with the non-FRNK plasmid group, the FRNK-transfected HSCs almost arrested in G0/G1 phase (71.4 +/- 2.81 vs 48.9 +/- 1.66, P < 0.01).</p><p><b>CONCLUSION</b>After FRNK were transfected successfully in HSCs using lipofectamine, the phosphorylation of FAK was inhibited. The HSC proliferation was restrained in a time-dependent manner and the HSC was arrested in G0/G1 phase.</p>

The role of FAK-ERK signal transduction pathway in apoptosis of hepatic stellate cell / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To investigate the effects of FAK-related non-kinase (FRNK) on the apoptosis of hepatic stellate cells (HSC) in vitro and on the extracellular signal-regulated kinase (ERK) signal transduction pathway.</p><p><b>METHODS</b>HSC were stimulated by fibronectin (FN), and then they were transfected with FAK-related non-kinase (FRNK) plasmids mediated by cationic liposome. The apoptosis of FRNK-induced HSC was examined by annexin-V/propidium iodide double-labeled flow cytometry (FCM), gel electrophoresis and transmission electron microscopy. Levels of FRNK, FAK, p-FAK (Tyr397), ERK1 and p-ERK in HSC were assayed by Western blot on the protein level, and by RT-PCR on the mRNA level.</p><p><b>RESULTS</b>The expression of FRNK was enhanced after FRNK plasmids were transiently transfected into the HSC. The apoptotic rate of the HSC exposed to FRNK plasmids for 48 h was higher than that in the non-FRNK plasmid group (25.37%+/-1.92% vs 9.28%+/-1.05%, P less than 0.01), and was accompanied by a significantly higher activity of caspase-3 both in the protein and in the mRNA levels [(264.17+/-12.60 vs 185.82+/-9.69), P less than 0.01; (4.19+/-0.48 vs 1.07+/-0.27), P less than 0.01]. After exposure of HSC to FRNK plasmids, compared with the non-FRNK plasmid group, the expressions of p-FAK, ERK1 and p-ERK in protein and mRNA levels were lower; on the contrary, compared with the control group, the expressions of p-FAK, ERK1 and p-ERK in the FN group were higher.</p><p><b>CONCLUSION</b>The expression of FRNK was enhanced and the phosphorylation of FAK was inhibited after FRNK was transiently transfected into HSC in vitro. FRNK induces apoptosis of HSC. FAK-ERK signal transduction pathway perhaps is involved in the process.</p>

The effects of FRNK on expressions of MMP-2 mRNA and TIMP-2 mRNA in hepatic stellate cells / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To investigate the effects of FAK-related non-kinase (FRNK) on expressions of type I collagen and matrix metalloproteinase-2 (MMP-2) mRNA and tissue inhibitor of metalloproteinase-2 (TIMP-2) mRNA in rat hepatic stellate cells (HSC).</p><p><b>METHODS</b>Using in vitro cell culture technique, FRNK plasmids were transfected into HSC mediated by cationic liposome. Type I collagen synthesis capability in HSC was examined by 3H-Pro incorporation assay. The levels of FRNK in HSC were assayed by Western blot, and the expressions of MMP-2 and TIMP-2 were assayed by RT-PCR on mRNA levels.</p><p><b>RESULTS</b>The exposure of HSC to FRNK caused the expression of FRNK protein to be up-regulated, and the FRNK protein contents reached the highest point at 48 h after the transfection, P less than 0.05. The expressions of MMP-2 mRNA were up-regulated by FRNK; the expressions of TIMP-2 mRNA were down-regulated by FRNK; the ratios of MMP-2 mRNA/TIMP-2 mRNA were enhanced by FRNK.</p><p><b>CONCLUSION</b>After FRNK was transfected, the capability of type I collagen synthesis in HSC was inhibited, which may be related to the up-regulation of MMP-2 mRNA/TIMP-2 mRNA.</p>

Comparative study of Buyang Huanwu Decoction and the different combinations of its ingredients on neurogenesis following ischemic stroke in rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effect of Buyang Huanwu Decoction (BYHWD) and the different combinations of its ingredients on neurogenesis following ischemic stroke in rats.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSION</b>BYHWD has a effect in promoting neurogenesis better than DGBXD.</p>

Effects of Buyanghuanwu decoction on nerve proliferation in rats with sequelae of ischemic stroke / 南方医科大学学报

<p><b>OBJECTIVE</b>To examine the effect of Buyanghuanwu decoction (BYHWD) in inducing nerve proliferation in rats with sequelae of ischemic stroke.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>The number of BrdU-immunoreactive cells in the cerebral tissues of BYHWD-treated rats was significantly greater than that in the untreated model group.</p><p><b>CONCLUSION</b>BYHWD can promote nerve proliferation in rats with ischemic stroke sequelae.</p>

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 / 中国中西医结合杂志

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>BHD could significantly inhibit the neuron apoptosis induced by hypoxia and down-regulate the expressions of p53 and p21 genes.</p><p><b>CONCLUSION</b>BHD shows inhibition on neuron hypoxia apoptosis and down-regulating of the p53 and p21 gene expression is one of its mechanisms.</p>

Study on preventive effect of buyang huanwu decoction on cardiomyocyte apoptosis induced by hypoxia-reoxygenation in rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>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).</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>