TNFAIP8 influences the motor function in mice after spinal cord injury (SCI) through meditating inflammation dependent on AKT.

Spinal cord injury (SCI) is a devastating disease and causes tissue loss and neurologic dysfunction, contributing to high morbidity and disability among human. However, the underlying molecular mechanisms still remain unclear. Tumor necrosis factor-α-induced protein 8 (TNFAIP8) is a member of the TNFAIP8/TIPE family, and has been implicated in different diseases associated with inflammation, infection, and immunity. Nevertheless, its effects on SCI have not been well investigated. In our study, we found time course of TNFAIP8 following SCI in mice, along with time-dependent increases of pro-inflammatory cytokines. The in vitro results confirmed the up-regulation of TNFAIP8 induced by lipopolysaccharide (LPS). Subsequently, we found that reducing TNFAIP8 by transfection with its specific siRNA (siTNFAIP8) markedly alleviated cell viability and inflammatory response caused by LPS in mouse microglial BV2 cells. Importantly, LPS-enhanced activation of inhibitor of κBα/nuclear factor-κB (IκBα/NF-κB) and phosphoinositide 3-kinase/serine-threonine kinase (PI3K/AKT) signaling pathways was considerably blunted by siTNFAIP8. Intriguingly, our results further showed that siTNFAIP8-restrained inflammation and IκBα/NF-κB in LPS-stimulated BV2 cells were almost abolished by the pre-treatment of AKT activator SC-79, demonstrating that TNFAIP8-regulated inflammatory response was largely dependent on AKT activation. Then, the in vivo studies were performed using the wild type (WT) and TNFAIP8-knockout (KO) mice with or without SCI operation. Results showed that TNFAIP8-KO mice exhibited improved neuron injury and locomotor function along with decreased microglial activity. Furthermore, compared with the WT/SCI mice, the expression of pro-inflammatory cytokines in spinal cords was markedly down-regulated by TNFAIP8-deficiency through blocking IκBα/NF-κB and PI3K/AKT signaling pathways. Taken together, these findings elucidated the novel role of TNFAIP8 in regulating SCI via the AKT signaling, and thus TNFAIP8 may be served as a promising therapeutic target for SCI treatment.

2,4-Dihydroxy-3'-methoxy-4'-ethoxychalcone suppresses cell proliferation and induces apoptosis of multiple myeloma via the PI3K/akt/mTOR signaling pathway.

Context: Caragana pruinosa Kom. (Fabaceae), a commonly used folk medicine, has been found to possess antitumor effects. However, the antiproliferative effect of 2,4-dihydroxy-3'-methoxy-4'-ethoxychalcone (DMEC) derived from C. pruinosa against multiple myeloma (MM) has never been investigated. Objective: This study systematically evaluates the antiproliferative effect of DMEC against MM cells. Materials and methods: The antiproliferative effect of DMEC (1, 2, 4, 8, 16, 32, and 64 µM) on MM cells lines, including RPMI8226, MM.1S, and U266, was examined using Cell counting kit-8 (CCK-8) assay after 24 h incubation. The proapoptotic effect of DMEC (20 µM) was determined using fluorescent microscope and flow cytometer, and its possible underlying mechanisms were further studied by using western blotting analysis. Results: The half maximal inhibitory concentrations (IC50) of DMEC on RPMI8226, MM.1S, and U266 cells were calculated as 25.97, 18.36, and 15.02 µM, respectively. The inhibitory effect of DMEC on MM cells was related to mitochondria-mediated apoptosis via upregulation of the cleaved-caspase-3 (C-3), cleaved-caspase-9 (C-9), Bad, and cytochrome C (Cyto C), but downregulation of the Bcl-2 and poly ADP-ribose polymerase (PARP). Furthermore, DMEC (5, 10, and 20 µM) reduced the expression of phosphatidylinositol-3-kinase (PI3K), phosphorylated (p)-protein kinase B (Akt), and p-mammalian target of rapamycin (p-mTOR), which were further evidenced by pretreatment with IGF-1, a PI3K activator. Conclusion: Collectively, our results indicate that the DMEC could be treated as a new candidate for treatment of multiple myeloma in the future. Also, an in vivo study is warranted in the future.

An effective treatment of experimental osteomyelitis using the antimicrobial titanium/silver-containing nHP66 (nano-hydroxyapatite/polyamide-66) nanoscaffold biomaterials.

Effective treatment of osteomyelitis remains a formidable clinical challenge. The rapid emergence of multidrug-resistant bacteria has renewed interest in developing antimicrobial biomaterials using antiseptic silver ions to treat osteomyelitis. However, inadequate local retention and severe cytotoxic effects have limited the clinical use of ionic silver for bone grafts. We recently developed novel porous nano-hydroxyapatite/polyamide 66 (nHP66)-based nanoscaffold materials containing varied concentrations of silver ions (Ag+) (TA-nHAPA66) and oxidized titanium (TiO2), which was added as a second binary element to enhance antibacterial activity and biocompatibility. In this study, we establish a large cohort of rabbit model of experimental osteomyelitis and investigate the in vivo antimicrobial and therapeutic effects of TA-nHP66 biomaterials and their in vivo silver release kinetics. We find the TA-nHP66 scaffolds exhibit potent antibacterial activities against E. coli and S. aureus, support cell adhesion and cell proliferation of pre-osteoblasts, and stimulate osteogenic regulator/marker expression. Moreover, the TA2-nHP66 scaffold exerts potent antibacterial/anti-inflammation effects in vivo and promotes bone formation at the lesion site of osteomyelitis. We further demonstrate that TA2-nHP66 exhibits excellent biosafety profile without apparent systemic toxicities. Therefore, the TA-nHP66 scaffold biomaterials may be further explored as an effective adjuvant therapy for infected bone defects and/or osteomyelitis debridement.

[PROTECTIVE EFFECT OF GLUCOSAMINE-HYDROCHLORIDE ON CARTILAGE IN BLOOD-INDUCED JOINT DAMAGE IN VIVO].

OBJECTIVE: To discuss the effect of glucosamine-hydrochloride (Glu/Ch) in protecting and repairing the cartilage in blood-induced joint damage (BJD) in vivo. METHODS: Thirty-two adult New Zealand rabbits were randomly divided into 4 groups (n=8):high-dose Glu/Ch treated group (group A), low-dose Glu/Ch treated group (group B), positive control group (group C), and negative control group (group D). A joint bleeding model was established by blood injection into articular cavity in groups A, B, and C. Glu/Ch was given by gavage in groups A (250 mg/kg) and B (21.5 mg/kg) once a day for 8 weeks, and the same dosage of saline was given in groups C and D. The serum cartilage oligomeric matrix protein (COMP), serum chondroitin sulfate 846(CS846), and urinary C-terminal telopepide of type II collagen (CTX-II) were measured at 3 days, 7 days, 2 weeks, and 8 weeks after modeling. The expressions of cytokines such as interleukin 1ß (IL-1ß) and tumor necrosis factor α (TNF-α) in synovial fluid were analyzed by ELISA at 8 weeks after modeling. The expression of matrix metalloproteinase 13(MMP-13) was detected by immunohistochemistry. Alcian blue staining and Safranin-O staining were performed to calculate the percentage of the positive staining areas. The proteoglycan content was detected by semi-quantitative analysis in the articular cartilage. RESULTS: The COMP concentration was significantly higher in groups A, B, and C than group D, and in groups B and C than group A at 3 days after modeling (P<0.05); no significant difference was found among groups A, B, and D at 7 days (P>0.05), and it was significantly lower in groups A, B, and D than group C (P<0.05); there was no significant difference among 4 groups after 2 and 8 weeks (P>0.05). Difference in CS846 concentration had no significance among 4 groups at each time point (P>0.05). The CTX-II concentration of groups A, B, and C was significantly higher than that of group D at each time point (P<0.05); it was significantly lower in group A than groups B and C at 7 days, 2 weeks, and 8 weeks (P<0.05). The TNF-α concentration of groups A and B was significantly higher than group D, and was significantly lower than group C at 8 weeks (P<0.05), but no significant difference was observed between groups A and B (P>0.05). The IL-1ß concentration was significantly higher in group C than the other groups (P<0.05), and in group B than groups A and D (P<0.05), but there was no significant difference between groups A and D (P>0.05). The MMP-13 expression was significantly higher in group C than groups A, B, and D (P<0.05), in groups A and B than group D (P<0.05). A significant decrease in the area stained with Alcian blue and Safranin-O was observed in group C. There were significant differences in the percentage of the positive stained areas of Alcian blue and Safranin-O among 4 groups (P<0.05). The relative quantities of proteoglycan from small to large in order was groups C, B, A, and D, respectively, showing significant differences (P<0.05). CONCLUSIONS: The metabolism disorder of cartilage matrix and synovium inflammatory reaction can be observed in rat joint bleeding model. Glu/Ch has certain protective effect on the cartilage after BJD by down-regulating IL-1ß, TNF-α, and MMP-13, as well as increasing proteoglycan content in the cartilage.

[BIOMECHANICAL STUDY ON KIDNEY-SHAPED NANO-HYDROXYAPATITE/POLYAMIDE 66 CAGE].

OBJECTIVE: To compare the biomechanical differences between the kidney-shaped nano-hydroxyapatite/polyamide 66 (n-HA/PA66) Cage and the bullet-shaped n-HA/PA66 Cage. METHODS: L2-L5 spinal specimens were selected from 10 adult male pigs. L2, L3 and L4, L5 served as a motor unit respectively, 20 motor units altogether. They were divided into 4 groups (n = 5): no treatment was given as control group (group A); nucleus pulposus resection was performed (group B); bullet-shaped Cage (group C), and kidney-shaped Cage (group D) were used in transforaminal lumbar interbody fusion (TLIF) through left intervertebral foramen and supplemented by posterior pedicle screw fixation. The intervertebral height (IH) and the position of Cages were observed on the X-ray films. The range of motion (ROM) was measured. RESULTS: There was no significant difference in the preoperative IH among 4 groups (F = 0.166, P = 0.917). No significant change was found in IH between at pre- and post-operation in group B (P > 0.05); it increased after operation in groups C and.D, but difference was not statistically significant (P > 0.05). There was no significant difference in the postoperative IH among groups B, C, and D (P > 0.05). The distance from Cage to the left margin was (3.06 ± 0.51) mm in group C (close to the left) and (5.68 ± 0.69) mm in group D (close to the middle), showing significant difference (t = 6.787, P = 0.000). The ROM in all directions were significantly lower in groups C and D than in groups A and B (P < 0.05), and in group A than in group B (P < 0.05). The right bending and compression ROM of group C were significantly higher than those of group D (P < 0.05), but no statistically significant difference was found in the other direction ROM (P > 0.05). CONCLUSION: The bullet-shaped and kidney-shaped Cages have similar results in restoring IH and maintaining the stability of the spine assisted by internal fixation. Kidney-shaped Cage is more stable than bullet-shaped Cage in the axial compression and the bending load opposite implant, it can be placed in the middle and back of the vertebral body more ideally.

Neuroprotective effect of Buyang Huanwu Decoction on spinal ischemia-reperfusion injury in rats is linked with inhibition of cyclin-dependent kinase 5.

BACKGROUND: Buyang Huanwu Decoction (BYHWD), a traditional Chinese medicine formula, has been shown to exert a variety of pharmacological effects including neuroprotective properties. However, the mechanism of neuroprotection is not fully understood. This study was designed to explore the mechanism of BYHWD in the treatment of spinal ischemia-reperfusion injury in rats. METHODS: Twenty-eight male Sprague-Dawley rats, weighting 250-280 g, were used, and were randomly divided into four groups with 7 animals in each: sham operation group (Control), spinal ischemia with saline (SI + Saline), spinal ischemia with BYHWD (SI + BYHWD), and spinal ischemia with roscovitine (SI + R). After 60 minutes of spinal ischemia followed by 72 hours of reperfusion, motor function of hind limbs, spinal ischemic infarction volume, the number of apoptotic cells, and cyclin-dependent kinase 5 (Cdk5) were examined. RESULT: Ischemia-reperfusion resulted in injury of the spines, while BYHWD significantly improved spinal function. The spinal infarction volume, number of apoptotic cells, and Cdk5 were decreased by administration of BYHWD. The similar improvements were seen with the pre-treatment of roscovitine. CONCLUSIONS: BYHWD prevented the ischemia-reperfusion-induced spinal injury in rats. The protective function of BYHWD was, in part, linked with inhibition of Cdk5.

Neuroprotective effect of Buyang Huanwu Decoction on spinal ischemia/reperfusion injury in rats.

AIM OF THE STUDY: The aim of this study was to investigate the protective effect of Buyang Huanwu Decoction, a traditional Chinese medicine formula, on spinal ischemia/reperfusion injury and explore the possible mechanism of the protective effect. MATERIALS AND METHODS: The spinal ischemia/reperfusion injury model was conducted in male Sprague-Dawley rats, and 40 g/kg Buyang Huanwu Decoction was administered by introgastric infusion. Motor function of hind limbs and apoptosis index were measured 72 h after reperfusion was started. The expression of thioredoxin and thioredoxin reductase was examined at 6h and at 24h after reperfusion. RESULTS: Motor function scores and apoptosis indices were significantly improved in the Buyang Huanwu Decoction group, as compared to the saline-infused control group. Spinal ischemia/reperfusion injury resulted in a decrease in the expression of thioredoxin, while Buyang Huanwu Decoction administration greatly elevated the expression of thioredoxin-1/thioredoxin-2 mRNA and thioredoxin reductase-1/thioredoxin reductase-2 mRNA. CONCLUSIONS: Our results suggest that administration of Buyang Huanwu Decoction may reduce spinal ischemia/reperfusion damage. This neuroprotective effect may be mediated, in part, by an increase in the transcription of thioredoxin.