| ?Objective?To investigate the effect of alcohol extract of Semen Cuscutae on postmenopausal osteoporosis and its possible molecular mechanism by detecting the expression levels of miR-21-5p and FasL in the femoral tissue of postmenopausal osteoporosis rats treated with alcohol extract of Semen Cuscutae.?Method?100 female SPF SD rats were randomly divided into 10 groups,including 10 rats of 2 months old and 90 rats of 6 months old,namely normal group of 2 months old,normal group of 6 months old,sham operation group,model group,estrogen group,alendronate sodium group,Semen Cuscutae extract group,low,middle and high dose group of Semen Cuscutae alcohol extract,except for the normal group of 2 months old,two weeks after adaptive feeding,were sampling.Other groups after 12 weeks of feeding,were sampling.The model group and the medication group were modeled by ovariectomy.The sham operation group only excised 1 cm3 of adipose tissue,and no excised was done.Except for the normal group of 2 months old,the rats in each group started to take 0.5ml/100 g of gastric perfusion volume from the second week after surgery,and the normal group of 6 months old and the sham operation group were given normal saline;low,middle and high dose group of Semen Cuscutae alcohol extract group,respectively,using Semen Cuscutae alcohol extract for gavage,dosing were 0.7g/kg,1.4g/kg,2.8g/kg;For the Semen Cuscutae extract group,the extract of Semen Cuscutae was used for intragastric administration,and the dosage was 0.35g/kg;Alendronate sodium was given to the alendronate sodium group at a dose of 6.3mg/kg.The estrogen group was given diethylstilbestrol at a dose of 0.0225mg/kg.The above 6-month-old group was administered with intragastric administration of No.16 gavage needle,and the alendronate group was intragastrically administered once a week.The other groups were intragastrically administered once a day.After 12 weeks of treatment,the samples were taken.At the 12 th weekend of drug use,3 rats in each group were selected for dual-energy X-ray bone density testing;After the experiment,the index of the uterus and the change of plasma alkaline phosphatase ALP were observed in each group.HE staining of femoral tissue to observe changes in trabecular bone;realtime-PCR was used to determine the relative expression of miR-21-5p in rat femur.Western blotting was used to determine the relative expression of FasL protein in rat femur.?Result?1.Uterine index: Compared with the normal group of 6 months old,the uterus index in the sham operation group had no significant change;compared with the sham operation group,the uterus index in the model group decreased significantly(P<0.05);compared with the model group,the uterus index in the estrogen group,Semen Cuscutae extract group,Semen Cuscutae alcohol extract low dose group,middle dose group and high dose group increased significantly(P<0.05);compared with the Semen Cuscutae extract group,the uterine index of rats in the Semen Cuscutae alcohol extract middle dose group had no significant change.2.Femur bone density: Compared with the normal group of 6 months old,the bone mineral density of femur in the sham operation group had no significant change;compared with the sham operation group,the bone mineral density of femur in the model group decreased significantly(P<0.05);compared with the model group,the bone mineral density of femur in the estrogen group,alendronate sodium group,Semen Cuscutae extract group,Semen Cuscutae alcohol extract low dose group,middle dose group and high dose group increased significantly(P<0.05);Compared with the normal group of 2 months old,the femoral bone mineral density of normal group of 6 months old decreased slightly,with no significant difference(P>0.05).3.Pathological morphology of the femur: Compared with the normal group of 6 months old,there was no significant change in the sham operation group;compared with the sham operation group,the trabecular bone of the model group was significantly thinner,the intervals widened,and there was no connection with each other.There were a few discontinuous,disordered free ends and irregular arrangement.Severe cancellous bone changes appeared in the cortex.Compared with the model group,the trabecular bone spacing of the estrogen group and the alendronate sodium group was widened,and the change of cancellous bone in the cortical area was not obvious.In the high dose group of Semen Cuscutae alcohol extract and Semen Cuscutae extract,the trabecular bone spacing was slightly wider,and the change of cancellous bone in cortical area was more obvious.In the low and middle dose groups of the alcohol extract of Semen Cuscutae,the trabecular bone spacing was widened and mild cancellous bone changes occurred in the cortical area.4.Effects on plasma alkaline phosphatase ALP: Compared with the normal group of 6 months old,the plasma alkaline phosphatase ALP content in the sham operation group had no significant change;compared with the sham operation group,the model group was slightly increased(P>0.05).Compared with the model group,the levels of plasma alkaline phosphatase ALP in the estrogen group,the alendronate sodium group,the Semen Cuscutae extract group and the Semen Cuscutae alcohol extract low,middle and high dose group of rats were increased(P>0.05);Compared with the normal group of 2 months old,the plasma alkaline phosphatase ALP content in the normal group of 6 months old rats was slightly decreased(P>0.05).5.Effect on the relative expression of miR-21-5p: Compared with the normal group of 6 months old,the relative expression of miR-21-5p in the sham operation group had no significant change;compared with the sham operation group,the relative expression of miR-21-5p in the model group was significantly increased,the difference was statistically significant(P<0.05).Compared with the model group,the relative expression levels of miR-21-5p in the femur tissues of the estrogen group,the alendronate sodium group,the Semen Cuscutae extract and the Semen Cuscutae alcohol extract low,middle and high dose group were decreased.And the estrogen group,the Semen Cuscutae alcohol extract low dose group and the middle dose group were significantly different,and the results were statistically significant(P<0.05).6.Effect on the relative expression of FasL protein: Compared with the normal group of 6 months old,the relative expression of FasL protein in the femoral tissue in the sham operation group had no significant change;compared with the sham operation group,the relative expression of FasL protein of the model group was significantly increased,and the results were statistically significant(P<0.05).Compared with the model group,the relative expression levels of FasL protein in the femur tissues of the estrogen group,the alendronate sodium group,the Semen Cuscutae extract group and the Semen Cuscutae alcohol extract low,middle and high dose group were decreased.And the estrogen group,the Semen Cuscutae alcohol extract low dose group and the middle dose group were significantly different,and the results were statistically significant(P<0.05).?Conclusion?The alcohol extract of Semen Cuscutae may down-regulate the expression of miR-21-5p in rat femur and up-regulate the level of FasL protein,thereby achieving the prevention and treatment of postmenopausal osteoporosis in rats. |
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