补肾健脾活血方对骨质疏松大鼠肝脏脂肪病变的作用

The effect of Bushen Jianpi Huoxue Fang on hepatic steatosis in osteoporotic rats

目的探索骨质疏松(osteoporosis,OP)模型大鼠的肝脏脂肪病变及补肾健脾活血方的作用。方法选取3月龄SPF级雌性大鼠24只,随机分为假手术组(n=6)、模型组(n=18);去卵巢后确定造模成功,随机抽取模型组12只大鼠,使用阿仑膦酸钠(n=6)和补肾健脾活血方(n=6)灌胃12周。取材后使用双能X线骨密度仪(dual-energy X-ray absorptiometry,DXA)检测骨密度(bone mineral density,BMD);microCT观察骨微结构;测量大鼠体重、肝脏重量及计算脂肪比;生化检测血清中谷丙转氨酶(alanine transaminase,ALT)、谷草转氨酶(aspartate transaminase,AST);油红O染色、HE切片观察肝组织;免疫组化观察谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)。结果与假手术组对比,模型组的ALT、AST水平升高(P<0.05),腰椎骨密度下降(P<0.05);骨小梁数量明显减少,骨微结构受破坏;体重升高,脂肪比升高;肝脏重量升高,肝脏脂肪浸润明显,肝小叶不清晰,肝索不规则,GPX4表达下降;与模型组对比,补肾健脾活血方组脂肪浸润减轻,肝小叶较模型组清晰,肝索较为规则,GPX4表达上升,P<0.05。结论OP可能导致肝的脂肪病变,补肾健脾活血方可减缓去势后大鼠肝脏脂肪病变的能力。GPX4可能在OP与肝脏脂肪病变的发生发展间产生一定的作用。

Objective To explore the hepatic steatosis in osteoporosis(OP)model rats and the effect of the nourishing kidney,invigorating spleen,and promoting blood flow recipe.Methods Twenty-four 3-month-old SPF-grade female rats were optionally allocated into the sham-operation group(n=6)and the mock-up group(n=18).After the ovaries were removed to ascertain the success of modelling,12 rats in the mock-up were randomly sampled,and the rats were gavaged with alendronate(n=6)and the nourishing kidney,invigorating spleen,and promoting blood flow recipe(n=6)for three months.BMD was detected using DXA.Micro CT was used to observe the bone microstructure.Biochemical tests were performed to detect ALT and AST.Oil-red O staining and HE sections were used to observe the changes in the liver.Immunohistochemistry was performed to observe the changes of glutathione peroxidase 4(GPX4).Results Compared to those in the sham group,ALT and AST in the mock-up group increased(P<0.05).BMD of the lumbar spine in the model group was lower than that in the sham-operated group(P<0.05).The number of bone trabeculae in the mock-up group was reduced and the bone microstructure was damaged.Fat infiltration in the liver was obvious in the mock-up group,and the hepatic lobes were unclear and the hepatic cords were far from regular.After intervention with the nourishing kidney,invigorating spleen,and promoting blood flow recipe,the fat infiltration was reduced and the hepatic lobes were clearer,and the hepatic cords were regular.Comparing to that in the mock-up group,the expression of GPX4 decreased in the model group(P<0.05).Conclusion Osteoporosis may also lead to fatty lesions in the liver.The nourishing kidney,invigorating spleen,and promoting blood flow recipe has the ability to slow down the fatty lesions in the liver of the rats.GPX4 may be involved in the relationship between osteoporosis and the progression of fatty liver.