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. 2016;39(1):85-92.
doi: 10.1179/2045772314Y.0000000273. Epub 2014 Oct 20.

Neuroprotective effects of the Buyang Huanwu decoction on functional recovery in rats following spinal cord injury

Dong Xian-Hui, He Xiao-Ping, Gao Wei-Juan

Neuroprotective effects of the Buyang Huanwu decoction on functional recovery in rats following spinal cord injury

Dong Xian-Hui et al. J Spinal Cord Med. 2016.

Abstract

Background: The Buyang Huanwu decoction (BYHWD) is a traditional Chinese herbal prescription and has been used in China to treat spinal cord injury (SCI) for hundreds of years. Clinical trials have shown that BYHWD improves the outcome of SCI in clinical trials, but the mechanisms are not known. This study observed the neuroprotective effects of BYHWD on spinal nerve cells after SCI and investigated possible mechanisms.

Materials and methods: Forty female Wistar rats were randomized equally to four groups treated by sham injury, SCI, BYHWD, or methylprednisolone (MP). The Basso, Beattie, and Bresnahan (BBB) score was used to evaluate hind-limb locomotor function. Neuron apoptosis was assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling staining and caspase-3, Bax, and Bcl-2 mRNA and protein expression were evaluated by real-time quantitative polymerase chain reaction and Western blotting, respectively.

Results: In the sham group, walking was mildly abnormal after anesthesia but recovered completely in 2 days. The BBB score in the SCI model group was significantly different from that in the sham group. The BBB scores of rats in both the BYHWD and MP groups were significantly higher than scores of rats in the SCI group. BYHWD had an antiapoptosis effect, as shown by significant decreases in expression of caspase-3 and Bax and increase in Bcl-2 expression.

Conclusion: BYHWD treatment restored hind-limb motor function of rats with SCI. The neuroprotective effect of BYHWD was associated with modulation of the expression of apoptosis-related proteins.

Keywords: Animal model; Apoptosis; BYHWD; Nerve cells; Spinal cord injury.

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Figures

Figure 1
Figure 1
Powdered R. astragali, R. angelicae sinensis, R. paeoniae rubra, R. chuanxiong, S. persicae, F. carthami, and Lumbricus. All materials were kindly provided by China Resources Sanjiu Medical and Pharmaceutical Co., Ltd. (A) R. astragali, (B) R. angelicae sinensis, (C) R. paeoniae rubra, (D) R. chuanxiong, (E) S. persicae, (F) F. carthami, (G) Lumbricus.
Figure 2
Figure 2
(A) Exposure of the spinal cord at T10–11 by laminectomy. (B) Special impactor equipment made in our laboratory. (C) The spinal cord after injury. (D) The injury site.
Figure 3
Figure 3
Effect of BYHWD on SCI alteration of BBB scores. At 1, 7, 14, 21, and 28 days after surgery, the BBB score of rats in the SCI model group were significantly different than scores of rats in the sham control group. The BBB scores of rats in the BYHWD and MP groups were significantly different than those of rats in the SCI group. n = 10 in each group; *P < 0.05 compared with the sham group; #P < 0.05 compared with the SCI model group.
Figure 4
Figure 4
Effect of BYHWD on SCI alteration of TUNEL assay. (A) Apoptotic cells in the sham group. (B) Apoptotic cells in the SCI group. (C) Apoptotic cells in the BYHWD group. (D) Apoptotic cells in the MP group. (E) Quantification of apoptotic cells. Significantly more apoptotic cells were observed in the SCI group (42.65%) than in the sham group (1.47%). There were significantly fewer apoptotic cells in the BYHWD (11.76%) and MP groups (10.29%) than in the SCI group. n = 4 in each group; *P < 0.05 compared with the sham group; #P < 0.05 compared with the SCI model group.
Figure 5
Figure 5
Effect of BYHWD on expression of caspase-3, Bax, and Bcl-2 mRNA by qPCR. BYHWD decreased caspase-3 and Bax mRNA expression but increased the expression of Bcl-2 mRNA. n = 6 in each group; *P < 0.05 compared with the sham group; #P < 0.05 compared with the SCI model group.
Figure 6
Figure 6
Effect of BYHWD on caspase-3, Bax, and Bcl-2 protein expression by Western blotting. BYHWD decreased expression of caspase-3 and Bax protein but increased the expression of Bcl-2 protein. n = 6 in each group; *P < 0.05 compared with the sham group; #P < 0.05 compared with the SCI model group.
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