[Effect of electroacupuncture on urodynamics and expression of Wnt-1, ß-catenin, and Ngn1 in the spinal cord in rats with bladder detrusor hyperreflexia due to supersacral spinal cord transection].

OBJECTIVE: To investigate the effect of electroacupuncture (EA) at "Dazhui" (GV14) and "Ciliao" (BL32) on rats with bladder detrusor hyperreflexia (DH) after supersacral spinal cord transection, as well as the mechanism of EA in improving the urinary function by regulating the expression of Wnt-1, ß-catenin and Neurogenin 1(Ngn1). METHODS: A total of 48 female Sprague-Dawley rats were randomly divided into sham-operation group, model control group, EA group, and EA control group, with 12 rats in each group. T10 spinal cord transection (SCT) was performed by surgery. The Basso, Beattie and Bresnahan (BBB) score was used to evaluate the motor function of SCT rat, and the Crede technique was used to assist urination. After the urine volume became stable, the urodynamic test was used to determine whether a rat model of DH was successfully established. The rats in the EA group were given EA at GV14 and BL32, and those in the EA control group were given EA (10 Hz/50 Hz, 20 min) at the acupuncture points at 1 cm next to GV14 and BL32 at both sides alternatively. EA was performed once a day for one week. Urodynamic parameters were used to evaluate urinary function. Western blot and immunohistochemistry were used to measure the expression of Wnt-1 and ß-catenin in the spinal cord, and immunofluorescence assay was used to measure the expression of Ngn1 in the spinal cord. RESULTS: The BBB score of the model control group significantly decreased compared with that of the sham-operation group(P<0.01), and the EA group was significantly higher than the model control group and the EA control group. Compared with the sham-operation group, the model control group had significant increases in bladder base pressure, maximum pressure, and leak point pressure (P<0.01) and significant reductions in maximum bladder capacity and compliance (P<0.01). Compared with the model control group, the EA group had significant reductions in bladder base pressure, maximum pressure, and leak point pressure (P<0.01) and significant increases in maximum bladder capacity and compliance (P<0.01, P<0.05). Compared with the EA group, the EA control group had significant increases in bladder base pressure, maximum pressure, and leak point pressure (P<0.01) and significant reductions in maximum bladder capacity and compliance (P<0.01, P<0.05). Compared with the sham-operation group, the model control group had significant increases in the protein expression of Wnt-1 and ß-catenin (P<0.05, P<0.01) and a signi-ficant reduction in the protein expression of Ngn1 in the spinal cord (P<0.01). Compared with the model control group, the EA group had significant increases in the protein expression of Wnt-1, ß-catenin and Ngn1 in the spinal cord (P<0.01). Compared with the EA group, the EA control group had significant reductions in the protein expression of Wnt-1, ß-catenin, and Ngn1 in the spinal cord (P<0.01). CONCLUSION: EA at GV14 and BL32 can significantly improve urinary function in rats with bladder DH due to SCT, partially by activating the Wnt/ß-catenin signaling pathway and promoting the protein expression of Wnt-1, ß-catenin and Ngn1.

Effect of kidney-reinforcing and marrow-beneficial Chinese medicine on bone metabolism-related factors following spinal cord injury in rats.

The present study aimed to investigate the effect of traditional Chinese kidney reinforcing and marrow-beneficial medicine (KRMB) on the prevention and treatment of abnormal bone metabolism and osteoporosis (OP) resulting from spinal cord injury (SCI). Rat models of OP following SCI were surgically established. The rats were randomly divided into five groups: Normal; sham operation + KRMB; normal + KRMB; SCI + KRMB; and SCI model group. Bone mineral density (BMD), and the expression of bone gamma-carboxyglutamic-acid containing protein (BGP), hepcidin mRNA and bone sialoprotein (BSP) were recorded at 1, 2, 4, 6, 8 and 10 weeks after the operation. BMD expression in the SCI model group was significantly lower compared with the normal, sham + KRMB and normal + KRMB groups at 4, 6, 8 and 10 weeks (P<0.01), and was significantly lower than that in the SCI + KRMB group at 6 (P<0.05), 8 and 10 weeks (P<0.01). The level of serum BGP in the SCI model group was significantly higher compared with the normal, sham + KRMB and normal + KRMB groups at each time point (P<0.01), and lower than the SCI + KRMB group (P<0.01). The SCI + KRMB group was significantly higher than the normal, sham operation + KRMB and normal + KRMB groups (P<0.01). Hepcidin mRNA expression in the rat livers in the normal, sham + KRMB and normal + KRMB group was significantly higher than that in the SCI + KRMB group and SCI model group at each time point (P<0.01). Hepcidin mRNA expression in the SCI + KRMB group was significantly higher than that in the SCI model group at 1 week (P<0.01), and significantly higher than the SCI model group at 2, 4, 6, 8 and 10 weeks (P<0.01). BSP expression in the SCI model group was significantly higher than that in the normal, sham + KRMB and normal + KRMB groups at each time point (P<0.01). BSP expression in SCI model group was higher than that in the SCI + KRMB group at 1 (P<0.05), 2, 4, 6, 8 and 10 weeks (P<0.01). In conclusion, KRMB traditional Chinese medicine may have a curative effect on secondary OP resulting from SCI.

Effect of kidney-reinforcing and marrow-beneficial traditional Chinese medicine-intervened serum on the proliferation and osteogenic differentiation of bone marrow stromal cells.

The present study aimed to investigate the effect of kidney-reinforcing and marrow-beneficial traditional Chinese medicine (TCM)-intervened (KRMBTI)-serum on the proliferation and osteogenic differentiation of bone marrow stromal cells (BMSCs) in rats. Rat BMSCs were isolated and cultured in vitro with various concentrations of serum obtained from rats at different time-points following treatment with low, medium and high doses of KRMBT. The alkaline phosphatase (ALP) activity and proliferation of the BMCSs was assessed to determine the optimal serum sampling time-point and serum concentration. Transforming growth factor (TGF)-ß1 expression of the BMSCs was detected using enzyme-linked immunosorbent assay (ELISA), and hepcidin mRNA expression in the rat livers was detected using reverse transcription polymerase chain reaction. The proliferation of BMCSs treated with serum obtained l h after dosing was observed to be significantly higher than that for BMCSs treated with serum obtained at the four other time-points (P<0.05). Furthermore, the proliferation following treatment with 25% KRMBTI-serum was significantly higher than that for the other KRMBTI-serum concentrations (P<0.01). For a 25% concentration of the serum collected at l h, the proliferation in the high- and low-dose KRMBTI-serum groups was significantly higher than that of the medium-dose and control groups (P<0.01) and no statistical significance was observed between the high- and low-dose groups. In the osteogenic differentiation process of the high-dose group, the ALP activity at every time-point was significantly higher than that of the low-dose group and the peak value of the former was achieved at concentrations between 20 and 30%. KRMBTI-serum was shown to promote the expression of TGF-ß1. Furthermore, hepcidin was observed to be expressed at significantly higher levels in the high-dose group than in the control group, and hepcidin expression was significantly higher after 10 weeks compared with that after five weeks. These findings suggest that KRMBTI-serum increases TGF-ß1 and hepcidin expression levels, which may be the mechanism underlying the promotion of osteogenic differentiation induced by KRMBTI-serum in BMSCs.