Antiosteoporotic activity of Du–Zhong–Wan water extract in ovariectomized rats

Figures & data

Figure 1. HPLC analysis of the plant extract reveals a complex composition of DZW water extract (Table 1).

Table 1. Characterization of DZW water extract compounds from HPLC chromatogram.

Retention time	Area (%)	Compound class	Compound name
3.861	2.532	Flavonoids	Isoferulic acid
9.498	4.727	Lignans	Geniposidic acid
10.589	2.075	Flavonoids	Oleuropein
13.692	2.798	Flavonoids	Cycloolivil
18.670	5.044	Phenylpropanoids	Chlorogenic acid
20.075	3.080	Lignans	Geniposide
20.913	2.454	Iridoid glycosides	Loganin
21.538	3.016	Organic acid	Caffeic acid
24.715	3.986	Lignans	Pinoresinol
25.873	2.616	Lignans	Pinoresinol diglucoside
27.082	1.879	Lignans	Dihydrodehydrodiconiferylalcohol
37.478	31.140	Iridoid glycosides	Eucommiol
38.508	2.711	Organic acid	3,4-Di-O-caffeoylquinic acid
40.997	2.455	Flavonoids	Astragalin
42.383	1.899	Organic acid	3,4-Dicaffeoylquininic acid
43.075	5.791	Triterpene Glycosides	Macranthoidin A
57.859	4.642	Triterpene Glycosides	Asperosaponin VI

Table 2. Body weight and uterus wet weight, serum parameters after 12 weeks administration of DZW (n = 10, mean ± SD).

Group	SHAM	OVX	E2	DZW – L	DZW – M	DZW – H
Body weight
Initial body weight (g)	272.500 ± 11.928	266.375 ± 8.105	272.375 ± 14.648	263.875 ± 7.200	269.585 ± 8.422	268.875 ± 9.643
End body weight (g)	300.750 ± 9.794	346.250 ± 12.163##	313.125 ± 19.172**	316.375 ± 13.999**	317.862 ± 11.396**	316.125 ± 13.747**
Uterus wet weight
Uterus wet weight (g)	0.404 ± 0.051	0.112 ± 0.012##	0.170 ± 0.012**	0.143 ± 0.018**	0.146 ± 0.015**	0.149 ± 0.017**
Serum parameters
OCN (pg/mL)	285.576 ± 39.005	217.949 ± 25.295##	248.458 ± 34.600*	251.169 ± 41.597*	254.174 ± 34.511*	258.119 ± 26.749**
TRACP-5b (U/L)	0.304 ± 0.026	0.375 ± 0.054##	0.266 ± 0.090**	0.283 ± 0.070**	0.277 ± 0.052**	0.247 ± 0.052**
Serum oestrogen (pmol/L)	2.702 ± 0.597	2.206 ± 0.334##	3.858 ± 0.612**	2.669 ± 0.502*	2.749 ± 0.408**	2.865 ± 0.699**

The data are expressed as mean ± SD, n = 10.

## p < 0.01 versus sham group,

* p < 0.05

** p < 0.01 versus the OVX group at the same time point as evaluated by ANOVA.

Table 3. BMD, Micro-CT properties of femoral trabeculae and biomechanical test of femur after 12 weeks administration of DZW (n = 10, mean ± SD).

Group	SHAM	OVX	E2	DZW – L	DZW – M	DZW – H
BMD (g/cm2)	0.265 ± 0.022	0.224 ± 0.007##	0.255 ± 0.007**	0.252 ± 0.011**	0.255 ± 0.009**	0.261 ± 0.019**
BV/TV (%)	58.470 ± 7.790	15.170 ± 2.880##	33.920 ± 3.870**	23.120 ± 4.210*	25.140 ± 3.780**	26.240 ± 3.970**
Tb.N (1/mm)	3.990 ± 0.150	2.040 ± 0.480##	3.030 ± 0.100*	2.760 ± 0.200*	2.760 ± 0.230**	2.790 ± 0.130*
Tb.Sp (μm)	101.230 ± 12.640	468.410 ± 69.420##	222.220 ± 10.740**	361.220 ± 34.940*	357.990 ± 29.490*	247.700 ± 54.350*
Tb.Th (μm)	161.550 ± 15.170	84.510 ± 8.730##	115.890 ± 9.100*	101.090 ± 6.480*	105.230 ± 7.810*	111.680 ± 11.220*
SMI	0.830 ± 0.070	2.930 ± 0.460##	1.510 ± 0.210**	2.090 ± 0.090*	2.040 ± 0.090*	2.020 ± 0.290*
maximum load (N)	87.329 ± 5.072	60.764 ± 3.165##	89.810 ± 5.656**	85.554 ± 11.158**	89.123 ± 9.375**	94.976 ± 13.169**
stiffness (N/mm)	193.538 ± 31.510	148.199 ± 10.667#	189.486 ± 15.733**	179.834 ± 18.773**	184.426 ± 16.125**	188.208 ± 16.327**
energy to ultimate load  (millijoules, mJ)	17.333 ± 2.582	12.000 ± 1.789##	18.667 ± 3.011**	25.833 ± 3.601**	27.479 ± 2.994**	28.167 ± 2.994**

The data are expressed as mean ± SD, n = 6.

# p < 0.05

## p < 0.01 versus sham group

* p < 0.05

**p < 0.01 versus the OVX group at the same time point as evaluated by ANOVA.

Figure 2. Representative Micro-CT images of trabecular bone microarchitecture in the distal femurs. (a) SHAM group, (b) OVX group, (c) E2 group, (d) DZW-L group, (e) DZW-M group and (f) DZW-H group. The OVX rats presented notable reduction in the trabecular number, trabecular area compared with the SHAM rats. DZW and E2 partially prevented OVX-induced trabecular bone loss and significantly improved trabecular bone mass and microarchitecture.

Figure 3. Representative ER expression of uterus in the endometrium. (a) SHAM group, (b) OVX group, (c) E2 group, (d) DZW-L group, (e) DZW-M group and (f) DZW-H group. The OVX rats presented notable reduction of ER labelling compared with the SHAM rats. DZW and E2 partially prevented OVX-induced ER expression reduction.