The Mechanism Of Zuoguiwan In The Prevention And Treatment Of PMOP By Regulating Leptin

ObjectiveObserving bone mineral density(BMD),the concentration of Leptin,βcross-linked C-telopeptide of type 1 collagen(β-CTX)and bone alkaline phosphatase(BALP)in serum,the expression of Leptin mRNA and protein in the hypothalamus,the expression of β2-adrenergic receptor(β2-AR),receptor activator of NK-κB ligand(RANKL),osteoprotegerin(OPG)mRNA and protein in the tibia of ovariectomized rats,to discover the possible mechanism of Zuoguiwan in the treatment of postmenopausal osteoporosis(PMOP)by regulating Leptin,and to supply theoretical and experimental references for other scholars.Methods50 female SD SPF rats were randomly divided into sham group and model group(with 10 and 40 rats).The way of 40 rats9 operation is ovariectomy.After conventional farming in a week,the remaining 36 ovariectomized rats were randomly distributed into model group,estradiol group,low dose Zuoguiwan’s group and high dose Zuoguiwan’s group.12 weeks intragastric administration later,take the femoral tissue to detect BMD.The 36 rates’ femoral morphological changes were detected by Hematoxylin and eosin(HE)staining.Leptin,p-CTX and BALP in serum were tested by enzyme-linked immunosorbent assay(ELISA).The mRNA changes of Leptin in the hypothalamus,β2-AR,OPG,and RANKL in the tibia were tested by qPCR.The protein changes of Leptin in the hypothalamus,β2-AR,OPG and RANKL in the tibia were tested by Western blot(WB).Result(1)BMD:The BMD of femur in the model group was lower than the sham group(P<0.01);compared with the model group,the femoral BMD in the low dose Zuoguiwan’s group,high dose Zuoguiwan’s group and the estradiol group was increased(P<0,01),and there is no obvious difference in the three groups(P>0.05).(2)Bone histomorphology:The bone trabecula observed by HE staining that in the model group with more interspaces,larger spacing,thinner volume,more destroyed structure,a larger number of empty bone lacuna,more irregularly arranged than sham group.Compared with the model group,the bone trabecula is denser,more,wider,with smaller space,more complete structure,less empty bone lacuna,more acceptable arrangement in low dose Zuoguiwan’s group,high dose Zuoguiwan’s group and the estradiol group(,P>0.05).(3)Serum detection index:Compared with the sham group,the serum Leptin and BALP of the model group were decreased(P<0.01),While the β-CTX was increased(P<0.01).Compared with the model group,the serum Leptin and BALP were increased,while β-CTX was decreased(P<0.01)in the low dose Zuoguiwan’s group,high dose Zuoguiwan’s group and the estradiol group,but there is no statistical difference in the three groups(P>0.05).(4)mRNA expression:Compared with the sharu group,the expression of Leptin mRNA in.the hypothalamus,RANKL and β2-AR mRNA in the tibia of the model group were increased(P<0.01;P<0.05),while OPG mRNA in the tibia was decreased(P<0.01);compared with the model group,Leptin mRNA in the hypothalamus,RANKL and β2-AR mRNA in the tibia were decreased,while OPG mRNA in the tibia of the low dose Zuoguiwan’s group,high dose Zuoguiwan’s group and the estradiol group were increased(P<0.01);compared with the estradiol group,β2-AR mRNA in the tibia of the low dose Zuoguiwan’s group and high dose Zuoguiwan’s group were decreased(P<0.01),OPG mRNA in the tibia of the low dose Zuoguiwan’s group was increased(P<0.01),there is no statistical difference in the other medicine intervention groups(P>0.05).(5)Protein expression:Compared with the sham group,Leptin protein in the hypothalamus and RANKL and β2-AR protein in the tibia of the model group was increased(P<0.05),while OPG protein in the tibia was decreased(P<0.05);compared with the model group,Leptin protein in the hypothalamus and RANKL and β2-AR protein in the tibia was decreased(P<0.01),while OPG protein in the tibia of the low dose Zuoguiwan’s group,high dose Zuoguiwan’s group and the estradiol group was increased(P<0.05),there is no statistical difference among the medicine intervention groups(P>0.05).Conclusion(1)Zuoguiwan can modulate molecular markers of bone turnover-β-CTX and BALP-and OPG/RANKL/RANK system to stimulate bone formation,restrain bone resorption and improve BMD.Control PMOP by adjusting Leptin in serum and hypothalamus two pathways.(2)One of the mechanisms of Zuoguiwan’s prevention and treatment of PMOP may be that affect OPG/RANKL/RANK system by increasing the level of serum Leptin so that increase the ratio of osteoblast and osteoclast.(3)One of the mechanisms of Zuoguiwan’s prevention and treatment of PMOP may be that affect OPG/RANKL/RANK system by decreasing the level of Leptin in hypothalamus and restraining the irritability of sympathetic nerve.(4)Zuoguiwan can tonify kidney,strengthen spleen,replenish essence and nourish the bone marrow to prevent and treat the deficiency of yin in women perimenopause and PMOP.