Colorectal cancer cells promote osteoclastogenesis and bone destruction through regulating EGF/ERK/CCL3 pathway.

Bone metastasis of colorectal cancer (CRC) cells leads to osteolysis. Aberrant activation of osteoclasts is responsible for bone resorption in tumor. In general, bone marrow-derived monocytes (BMMs) differentiate into osteoclasts, however, how CRC cells interact with BMMs and how to regulate the differentiation is elusive. We here report that CRC cells promote bone resorption in bone metastasis. Transcriptomic profiling revealed CCL3 up-regulated in MC-38 conditional medium treated BMMs. Further investigation demonstrated that CCL3 produced by BMMs facilitated cell infusion and thus promoted the osteoclastogenesis. In addition, CRC cells derived EGF stimulated the production of CCL3 in BMMs through activation of ERK/CREB pathway. Blockage of EGF or CCL3 can efficiently attenuate the osteolysis in bone metastasis of CRC.

Effect of recombinant human thioredoxin on Coxsackie virus 3m-induced cell injury / 中国地方病学杂志

Objective To observe the protective function of recombinant human thioredoxin(TRX) on HeLa cell injury induced by Coxsackie virus 3m(CVB3m) and to study the inhibiting effect of TRX on viral replication. Methods We infected HeLa cells with 10TCID50 CVB3m and then protected these cells with TRX (2,5,10 mg/L). The protective group of TRX, viral group, control group of TRX, and normal control group were included. Six parallel wells were set up in each group. The cell growth was observed by methyl thiazolyl tetrazolium(MTT) and contrast phase microscope. Results The results of contrast phase microscope revealed that HeLa cells were arranged tightly and polygon in normal control group; untightly, became circle and abscission in viral group; HeLa cells morphous improved by increasing TRX concentration in TRX protective group(2,5,10mg/L). MTT results of the inhibitory ratio on cell growth of TRX(2,5,10 mg/L) control group(1.2%,2.9%,6.3%) were compared with normal control group(0), there was no significant difference(all P > 0.05); and while the inhibitory ratio on cell growth of TRX(2,5,10 mg/L) protective group(32.0%,28.0%,27.0%) was compared with virus infective group(51.7%), there was a significant difference (all P < 0.05). The inhibition study of viral replication showed that compared the inhibitory ratio on cell growth of TRX(2,5,10 mg/L) protective group(26.0%,27.0%, 10.9%) with virus infective group(60.0%), there was a significant difference(all P < 0.05). In the protective groups, there was a significant difference (all P < 0.05) between low dose groups(2,5 mg/L) and high dose groups( 10 mg/L). Conclusions The recombinant TRX(2,5,10 mg/L) may alleviate HeLa cell's injury induced by virus and the construct has no significant toxicity. TRX(2,5,10 mg/L) is effective in inhibiting virus CVB3m replication.

Human thioredoxin exerts cardioprotective effect and attenuates reperfusion injury in rats partially via inhibiting apoptosis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Thioredoxin is one of the most important redox regulating proteins. Although thioredoxin has been shown to protect cells against different kinds of oxidative stress, the role of thioredoxin in myocardial ischemia and reperfusion injury has not been fully understood. This study was conducted to explore the protective role of human thioredoxin on myocardial ischemia and reperfusion injury and its potential mechanisms.</p><p><b>METHODS</b>Purified human thioredoxin was injected into adult Wistar rats, which were subjected to 30 minutes of myocardial ischemia followed by 2 or 24 hours of reperfusion. We detected 1) the infarct size; 2) the level of malondisldehyde (MDA) in serum; 3) the expression of caspase-9, and cytochrome c in/out of mitochondria by Western blotting; 4) apoptosis by terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) assay and caspase-3 and its protein by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blotting; 5) the expression of bcl-2 and bax in cardium by immunohistochemical (IHC) assay.</p><p><b>RESULTS</b>Human thioredoxin reduced myocardial ischemia/reperfusion injury as evidenced by significant decrease of myocardial infarct size (P < 0.01), notable reduction of myocyte apoptosis (P < 0.01), lower systemic oxidative stress level (P < 0.01) after reperfusion for 2 hours, and few inflammatory cell infiltration after reperfusion for 24 hours in rats. Furthermore, treatment with human thioredoxin significantly reduced the release of mitochondrial cytochrome C (P < 0.05), and inhibited the activity of caspase-9 (P < 0.05) and caspase-3 (P < 0.01 in mRNA and P < 0.05 at protein level). Meanwhile, human thioredoxin markedly increased bcl-2 expression (P < 0.05).</p><p><b>CONCLUSIONS</b>These results strongly suggest that human thioredoxin has cardioprotective effects on myocardial ischemia/reperfusion and its anti-apoptotic role may be mediated by modulating bcl-2 and the mitochondria-dependent apoptotic signaling pathway.</p>

Effect of insulin on the cognizing function and expression of hippocampal Abeta1-40 of rat with Alzheimer disease / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>A model of simulated Alzheimer's disease (AD) induced by aggregated amyloid protein (Abeta(1-40)) was built in Wistar rats to observe the behavioral and pathological changes of Abeta(1-40) and the effect of hypodermic insulin injected on the function of study and memory and the expression of Abeta(1-40) from the CA1 area of the hippocampus.</p><p><b>METHODS</b>Experimental groups were as follows: contrast, simulated AD model, contrast of Nacl, and insulin treated. The simulated AD model was built by microinjection of aggregated Abeta(1-40) at the CA1 area of the hippocampus, and was hypodermically injected with 0.9% NaCl (1 ml/kg) and insulin (0.1 U/kg) separately the next day. Two weeks after the modeling, the four groups were tested with water maze about the study and memory function of rats. Three weeks after the injection, the expression of Abeta(1-40) at the CA1 area of the hippocampus was examined by pathological tests (HE, Congo red) and immunohistochemical methods.</p><p><b>RESULTS</b>The study and memory abilities of rats were ameliorated significantly by the place navigation test and the spatial probe test after the application of insulin. Insulin could decrease the expression of Abeta(1-40) at the CA1 area of the hippocampus to reduce the pathological damage of Abeta(1-40) to the hippocampal area of rats.</p><p><b>CONCLUSIONS</b>The injection of aggregated Abeta(1-40) to the hippocampal area could simulate the behavioral and pathological features of AD such as the difficulty of study and memory and the damage to neurons. Insulin is effective to improve the function of study and memory and amend the pathological damage of simulated AD model rats. The results give a experimental proof of insulin in the clinical treatment of AD.</p>

Protective effects of recombinant human thioredoxin on myocardium in mice with viral myocarditis / 中国地方病学杂志

Objective To evaluate protective effects of recombinant human thioredoxin(TRX) in myocardial injury of mice with viral myocarditis. Methods We established viral myocarditis models by intraperitoneal injection with 0.1 ml 100TCID50 Coxsackie virus 3m(CVB3m), along with tail vein injection of recombinant human TRX (2 mg/kg) for protection. The control group was given equivalent volume of normal saline. The mice were killed 7 days following the injections. Serum lactate dehydrogenase (LDH) activity was determined and myocardial injury was examined with light microscopy. Results The somm LDH activity in Coxsackie virns-infected mice [(3130.50±390.57)U/L] was higher than that of animals in the control group[ (1617.86±155.42)U/L] and that of TRX protection group[ (1959.43±540.75)U/L], the difference being statistically significant (P<0.05); there was no significant difference between TRX protection group and the control group(P 0.05). Light microscopy showed that five of the eight Coxsackie rims-infected mice had myocardial lesions, including focal myocardial necrosis and inflammatory infiltration. There was no myocardial injury in the TRX protection group. Conclusions Recombinant human TRX can lessen myocardial injuries induced by infection with CVB3m, and so can protect myocardium.

Effects of resveratrol on isolated thoracic aorta rings of rats / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the relaxative characteristics of resveratrol on thoracic aortic artery in the rat and its mechanism.</p><p><b>METHOD</b>We perfused the isolated rings and observed the response of NA-induced artery contraction to resveratrol under the Ca2+-contained and Ca2+-free bath solutions. In the same way were the effect of reveratrol on the vascular smooth muscle observed by adding two different concentration of KCl (30 and 80 mmol x L(-1)), and the effect on the contraction of the vascular smooth muscle depending on the intracellular calcium and extracellular calcium were also observed by adding NA. We also observed the effect of resveratrol on the contraction of rings induced by NA in the presence of L-NNA and Glibenclamide.</p><p><b>RESULT</b>Resveratrol relaxed rat aorta rings precontracted by NA in a dose-dependent manner. The relaxant effect of resveratrol on the rat rings of endothelium-denuded group was reduced compared with that of endothelium-intact group; the relaxant effect of resveratrol on rat rings was higher under the condition of Ca2+-free bath solution than that under the condition of Ca2+-contained bath solution. Resveratrol had a repressive effect on the aorta's contraction induced by intracellular calcium, but had no effect induced by extracellular calcium. Resveratrol relaxed the contractions induced by KCl 30 mmol x L(-1) as well as KCl 80 mmol x L(-1), but the contraction curve of KCl 80 mmol x L(-1) was shifted upward significantly. In the L-NNA group, the relaxant effect was attenuated by (26.0 +/- 4.6) %; but there was no change in the group of Glibenclamide ( P > 0.05).</p><p><b>CONCLUSION</b>The results indicate that resveratrol relaxes vascular smooth muscle in an endothelium dependent manner. The mechanisms for this phenomenon seem to be related with promoting synthesis and release of NO, opening Ca2+ activated K+ channel (KCa channel) as well as the inhibition of Ca2+ influx and release of Ca2+ from intracellular stores.</p>