骨疏康对1型糖尿病骨质疏松大鼠体外培养的破骨细胞影响

Effect of decoction for treatment of osteoporosis on osteoclasts derived from bone marrow

目的:观察骨疏康对1型糖尿病骨质疏松大鼠体外培养破骨细胞的影响。方法:雌性Wistar大鼠60只,2.5～3月龄,按照体重随机分为正常组(n=24)和1型糖尿病组(n=36)两大组,正常组又分为正常对照组(n=8)、正常假手术组(n=8)和正常双侧卵巢切除组(n=8);1型糖尿病组又分为糖尿病对照组(n=12)、糖尿病假手术组(n=12)和糖尿病双侧卵巢切除组(n=12)。单剂量腹腔注射链脲菌素(柠檬酸钠缓冲液制成2％溶液) 60 mg·kg-1制备1型糖尿病大鼠模型;无菌条件下切除大鼠双侧卵巢制备骨质疏松模型。在造模后第0,2,4,8周末时用sRANKL和巨噬细胞集落刺激因子(M-CSF)诱导大鼠骨髓进行体外破骨细胞培养,并观察骨疏康(20μg·mL-1)的影响。结果:糖尿病对照组破骨细胞明显高于对照组(P<0．01),糖尿病双侧卵巢切除组破骨细胞明显高于对照组(P<0．01),糖尿病骨质疏松组破骨细胞明显高于糖尿病对照组和正常双侧卵巢切除组(P<0．01)。经20μg·mL-1骨疏康干预后,各组破骨细胞数均明显减少(P<0．01)。结论:骨疏康明显抑制1型糖尿病骨质疏松大鼠体外培养的破骨细胞生成。

Objective;To assess the effect of decoction for treatment of osteoporosis （ Gushukang） on osteoclasts derived from bone marrow of rats with type 1 diabetic osteoporosis and cultured in vitro. Methods ： Sixty 2.5 to 3-month-old female Wistar rats were randomly assigned to one of two categories： normal control （NC, n = 24） or type 1 diabetic control （ DC, n = 36）. Each category was then randomly 1 ： 1 sub-grouped into control （ NC and DC） , sham-ovariectomy （ NS and DS） or ovariectomy （ NOVX and DOVX） , respectively. The type 1 diabetic model of rats was set up by abdominally injecting a single dose of streptozotocin 60 mg· kg^-1, and the osteoporosis model of rats was induced by ovariectomy. The osteoclasts derived from bone marrow cells of the rats were induced by co-cultures of sRANKL and M-CSF in vitro at 0, 2, 4, 8 weeks post the ovariectomy, and Gusgukang 20 g·mL^-1 was simultaneously added into the cultures to evaluate its effect for the osteoclast formation. Results：The number of osteoclasts in diabetic group was significantly higher than the one in the normal control group. The Gushukang-treated rats experienced the less osteoclasts （ P 〈 0.01 ）. Conclusions ： Kushukang functioned to inhibit the osteoclasts formation in type 1 diabetic osteoporosis of rats in vitro.