[Effects of emodin on oxidative stress and inflammatory response in rats with acute spinal cord injury].

This paper was aimed to investigate the effects of emodin on oxidative stress and inflammatory response in rats with acute spinal cord injury（SCI）, and to explore the protective mechanism of emodin on neurons after SCI. Rat SCI models were established using a modified Allen's method. One hundred and ninety five Sprague-Dawley rats were randomly divded into sham (group A), model (group B), emodin group of 20 mg·kg⁻¹(group C), emodin group of 40 mg·kg⁻¹(group D), emodin group of 80 mg·kg⁻¹(group E). Functional recovery was evaluated using the Basso-Beattie-Bresnahan (BBB) scale and inclined plate test on the 3rd, 7th, 14th and 28th day. On the 7th day after SCI, neuron nylon body was observed with toluidine blue staining. The changes of myelinated nerve fibers were observed by transmission electron microscope. Nrf2, HO-1, NQO1, GFAP, NF-κB protein were detected by Western blot. The content of TNF-α, IL-1ß, IL-6 were detected by ELISA. Immunofluorescence staining was performed to observe the expression of the GFAP, NG2 and ED-1. The BBB and inclined plate scores of group C, D and E were higher than group B on the 7th, 14th, 28th day,the difference was statistically significant (P<0.05). On the 7th day, Nylon Body of group B and C started to fuse,the fusion of group D and E were significantly alleviated than group B and C. Transmission electron microscope showed that the changes of demyelination were obvious in group B and C, group D and E were significantly improved than group B and C. Western blot showed that Nrf2, HO-1, GFAP, NF-κB protein expression,group C, D, E compared with group B, NQO1 protein expression, group D, E compared with group B, the difference was statistically significant (P<0.05). ELISA showed that the content of TNF-α, IL-6,group C, D, E compared with group B,the content of IL-1ß,group D, E compared with group B, the difference was statistically significant (P<0.05)；Immunofluorescence showed that the expressions of GFAP and NG2 in group C, D and E were higher than that in group B, and the group D and E were more obvious. The expression of ED-1 in group C, D, E were decreased significantly compared with group B. Emodin has protective effect on neurons after SCI. The mechanism may connect with activting Nrf2-ARE pathway, reducing the expression of NF-κB, ED-1, TNF-α, IL-1ß, IL-6, and promoting the expression of GFAP and NG2.

[Effects of tanshinone â ¡A sulfonate on nerve regeneration after cryopreserved sciatic nerve allograft in rats].

In order to investigate the protective effect of tanshinone ⅡA sulfonate on the sciatic nerve activty in rats after cryopreservation as well as the nerve regeneration and functional recovery after allograft and its possible mechanism, Sprague-Dawley (SD) rats were divded into four groups at different doses of tanshinone ⅡA sulfonate (A 0 mg·L⁻¹, B 80 mg·L⁻¹, C 160 mg·L⁻¹, D 480 mg·L⁻¹) cryopreserved at -80 °C for 24 weeks. Fresh control group nerve segments were harvested without cryopreservation. The ultrastructure and the viable cells of the nerve segments after cryopreservation were observed by electron microscopy, calcein-AM/propidium iodide staining, respectively. The expression of Bax and Bcl-2 was detected by Western blot. After cryopreservation, the nerve segments were cultured in vitro for one week, the mRNA and protein level of NGF and GDNF were detected by PCR and Western blot respectively. In addition, the above four cryopreserved groups transplanted to the Wistar rats by allografting (A', B', C', D'). At 16-week postoperation, muscle compound action potential latency and nerve conduction velocity were examined by electrophysiological. The number and the thickness of myelinated nerve fibers were analyzed by toluidine blue staining. The ultrastructure of the sciatic nerve by electron microscopy was observed. According to the results, after the cryopreserved for 24 weeks, compared with groups A and B, the nerve demyelination and vacuolation were weak, and the more viable cells, the decreased Bax and increased Bcl-2, the increased NGF and GDNF in group C and D. At 16-week poseoperation, the results demonstrated that the more larger and thickly regenerated myelinated axons, the shorter latency of muscle compound action potentials and higher nerve conduction velocity in groups C' and D' compared with groups A' and B'. According to these results, tanshinone ⅡA sulfonate exerted a significant protective effect on the viability of the nerves during cryopreservation at -80 °C and promoted nerve regeneration and functional recovery after transplantation especially in middle- and high-dose of tanshinone ⅡA sulfonate.

[Effects of electroacupuncture on synovia IL-1beta and TNF-alpha contents in the rabbit with knee osteoarthritis].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on synovia IL-1beta and TNF-alpha contents in rabbits with knee osteoarthritis (KOA). METHODS: A total of 40 New Zealand rabbits were randomly divided into control, model, massage and EA groups. KOA model was established by gypsum fixing method. EA (2 Hz, 1-3 mA) was applied to left "Xiyan" (EX-LE 5), "Yanglingquan" (GB 34), "Xuehai" (SP 10), "Zusanli" (ST 36) and "Liangqiu" (ST 34) for 30 min, once daily and continuously for 21 days. For massage group, the affected knee joint was pressed, kneaded, stretched and rotated repeatedly for 15 min every time, followed by forced running about 100 m. The intra-joint synovia was collected (0.4-0.6 mL) for detecting the contents of IL-1beta and TNF-alpha with radioimmunoassay. RESULTS: No IL-1beta and TNF-alpha were detected in the synovia in control group, while in the other 3 groups, synovia IL-1beta and TNF-alpha levels increased significantly. Before treatment, no significant differences were found among model, EA and massage groups in the levels of IL-1beta and TNF-alpha (P > 0.05), while after the treatment the two indexes both decreased considerably (P < 0.05). Compared with model group, the two indexes were remarkably lower (P < 0.05), but no significant differences were found between EA and massage groups in synovia IL-1beta and TNF-alpha contents (P > 0.05). CONCLUSION: Both EA and massage can effectively suppress the release of synovia IL-1beta and TNF-alpha in KOA of rabbits, which may contribute to the effect of acupuncture in the treatment of osteoarthritis.