| Purpose:To research the effect of Zuogui Pill on preventing and curing postmenopausal osteoporosis (PMOP),through the PMOP modeled rats Ovariectomized. Consequently,to dwas cuss the therapeutic mechanwas m on signal transduction pathway of TGF-β/Smad. All those practice have been made on baswas of the theory of kindey storing the body essence and in charge of bone and generating marrow. Material and method:Divided 91 SD female rats at 4.5 months to 3 groups temporarily:Normal group with 13 rats,Sham group with 13 rats,for the rest 65 rats were eccoped both ovaries to establish PMOP models. 21 days later,divided the model rats into 5 groups randomly:OVX,N,ZG(H),ZG(M),ZG(L),13 rats per group. ZG(H),ZG(M), ZG(L) were ig. with Zuogui Pill suspl.(high,middle,low dose),N was ig. with Nilestriol as postive control group,Normal,Sham,OVX were ig. with dwas tilled water. Rats were weighten per week to regulate the dose. 120d later,except the dead rats during ig.,84 rats were cut off food and water for 24h,then killed,collected blood from abdominal aorta that was standing for 2h and centrifuged to get serum to test hebin level,stored at -70℃,the left hind limb at the 1/3 of dwas tal femur was taken to test test the expression of TGF-β1,Smad4 mRNA in bone tissues,which should be removed soft tissue and fascia,than treated by DEPC and packaged with aluminium foil,quick freezed in liquid nitrogen and stored at -70℃;The femoral bone of right hind limb was taken to determine the bone mineral density (BMD),which should be removed muscls and fascia but peritosteum,water flushed and packaged with saline water sinked gauze in refreshing bags,stored at -20℃.Results:1. The effect of Zuogui Pill on morphometry parameter of bone(1) Histomorphology of bone tissue Under the light microscope, the bone trabecula of Normal, Sham is complete, close linked in order,net shaped with small cavitas medullaris; bone trabecula of OVX changes to be disorder,thinner,relatively sparse,lack of connection,and with more fracture;the medicine treated rats of N,ZG(H),ZG(M),ZG(L) has more hyperplasy,linked in order and strong,specially the N and ZG(L).(2) Morphometry of bone tissueCompared with Sham, the %Tb·Ar and Tb·Th of OVX rats decreased,Tb·Sp increased significently(P<0.01);compared with OVX,%Tb·Ar and Tb·Th of medicine treated rats increased,and Tb·Sp decreased significantly(P<0.01) without differences between every two groups(P>0.05).2. The effect of Zuogui pill on the BMD of right hind limb at the 1/3 of proximate caput femorisCompared with Sham, the BMD of proximate caput femoris of OVX rats decreased significantly (P<0.01);compared with OVX,the the BMD of proximate caput femoris of medicine treated rats increased significantly (P<0.01),without differences between every two groups (P>0.05).3. The effect of Zuogui Pill on bone metabolism(1) The effect on BGPCompared with Sham,BGP of OVX increased significantly(P<0.01);compared with OVX,BGP of medicine treated rats decreased significantly(P<0.01),specially the ZG(M),ZG(L) but differences (P>0.05).(2) The effect on TRAPCompared with Sham,TRAP of OVX increased significantly (P<0.01);compared with OVX, BGP of medicine treated rats decreased significantly (P<0.01),without differences between every two groups(P>0.05).4. The effect of Zuogui Pill on sex hormonesCompared with Sham,E2 of OVX decreased significantly(P<0.01),FSH,LH increased significantly(P<0.01); compared with OVX,E2 of medicine treated rats increased(P<0.05),and FSH,LH increased(P<0.05) without differences between every two groups. 5. The effect of Zuogui Pill on expression of TGF-β1,Smad4 mRNACompared with Sham, expression of TGF-β1,Smad4 mRNA of OVX decreased significantly(P<0.01);compared with OVX,the expression of medicine treated rats increased significantly(P<0.01),without differences between every two groups(P>0.05).Conclusion:1. Eccope the both ovaries of rats can establish the PMOP models.2. Zuogui Pill can prevent and cure PMOP effectively.3. The down regulation of TGF-β1 and Smad4 mRNA in bone tissues may be the one important machanism of PMOP attacking.4. Zuogui Pill can up regulate the TGF-β1 and Smad4 mRNA in bone tissues, rebalance the bone metabolism, and that may be related to the mechanism of therapy action. |
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