Complex Fangxian Zhen Tang Balance In Postmenopausal Women With Osteoporosis And Its Impact On Osteoblasts

Osteoporosis is one of most frequent diseases of abnormal general bone metabolism at present.As the disease often incorporates fractures of wrist,lumbar and hip,it does harm for human beings,particularly for women and older people.Fall is one of reasons causing fracture.Because the balance ability of elderly people decreases,the rates of fall increases.Chinese herbs have good effects to osteoporosis.Compound Xian Zhen Decoction has been attributed to modify osteoporotic symptoms and increased bone mineral density(BMD).Further researching Compound Xian Zhen Decoction how to treat osteoporosis and anendment the balance ability will give new way to treat osteoporosis.PURPOSE:1.In order to evaluate the effect of preventing falls with Compound Xian Zhen Decoction ,The clinic trail was designed to observe the effect to muscle strength and balance ability in postmenopausal women suffered with osteoporosis.2. In order to investigate the mechanism of action of treating osteoporosis from cell molecular level, the cell experiment was designed to cultivate osteoblast with Compound Xian Zhen Decoction,then test the cell proliferation rate by MTT colorimetric assay and apoptotic proportion by Flow Cycometry (FCM) respectively .METHODS:Clinic trail:Randomized controlled trail was designed. A total of 96 postmenopausal women suffered with osteoporosis, aged 50-80 years and lived in Beijing,were randomly divided into four groups,treatment group,western control group,Chinese herb control group,and Sport group.All groups participated in taking a walk with gait speed of 120 steps per minute 3 days per week. Walking lasted 30 minutes every time.In addition, treatment group received Compound Xian Zhen Decoction one dosis daily,western control group received salmon calcitonin nasal spray 20μg and Caltrate 600 mg daily,Chinese herb control group received Xian Ling Gu Bao Capsule 6 grains/24 hours.All participants were prohibited to receive estrogen or diphosphonate during trails.The course of treatment was 6 months.Balance ability (including walking test on balance beam, timed up and go test, one-leg balance test with eyes closed , strengthening Romberg test),testosterone and estradiol in serum, BMD of lumbar and neck of femur,symptoms were measured before and after treatment.Cell experiment: We cultivated osteoblast strain UMR106 with Minimum Essential Medium (MEM) contained 10% Fetal bovine serum until the osteoblasts spread a thin layer at the bottom of the bottle. Then we digested the cells and adjusted its concentration to 2 x104/ml.The next step, we inoculated the cells into 96 wells of the tissue culture plate.After that,we added the Compound Xian Zhen Decoction whose concentration were 1 mg/L,5 mg/L,10 mg/L,50 mg/L, 100 mg/L,500 mg/L,1000 mg/L respectively to wells.After the osteoblast cultivated for 72 hours, the proliferation rate were tested by MTT colorimetric assay.When apoptosis was observed,osteoblast were cultivated as the same as above method.Then they were inoculated into the tissue culture plate with 6 wells and cultivated by MEM without fetal bovine serum for 14 hours. Next, the osteoblast were divided into four groups: Group 1 was the placebo group and received MEM without fetal bovine serum, Group 2 ,as the model group, received TNF-a 30ng/ml of concentration. Group 3 ,as treatment groups ,divided further into four groups which all received with TNF-a (30ng/ml of concentration) and at the same time received Compound Xian Zhen Decoction whose concentration was 500mg/L, 50mg/L, 10mg/L 1mg/L respectively. Then they were cultivated for 72 hours. At last, after the osteoblast were fixed by 70% icy ethanol and colored by propidium idodide alone, the apoptotic proportion was tested by the flow cycometry.RESULTS:Clinic trail:Balance abilities test:In Compound Xian Zhen Decoction group, time decreased significantly in timed upand go test ( P<0.05 ) and lengthened significantly in strengthening Romberg test(P<0.05)comparing to before treatment.There were no significantly changes among other groups.BMD:Comparing with pretherapy, BMD of lumbar was increased significantly but BMD of neck of femur was no change in Compound Xian Zhen Decoction group in post-treatment.But BMD of lumbar and neck of femur increased significantlyin salmon calcitonin group(both P<0.01).Furthermore . There were no significant difference between Compound Xian Zhen Decoction group with salmon calcitonin group(P>0.05).All BMD of Xian Ling Gu Bao Capsule group and Sport group was no significant change.Testosterone and estradiol:Comparing with prior treatment,testosterone and estradiol were no significant change in all groups in post-treatment. Symptoms:Compound Xian Zhen Decoction had best effect to modify symptoms and was significant difference with orther groups.Cell experiment:Compound Xian Zhen Decoction could promote osteoblast to proliferate and 500mg/L concentration showed the best function(P<0.01﹚.The osteoblast apoptosis proportion induced by TNF-a decreased significantly in Compound Xian Zhen Decoction of 500mg/L and 1mg/L concentration group comparing with model group ( both P < 0.01).1mg/L concentration group played better role.Compound Xian Zhen Decoction of 500mg/L,10mg/L and 1mg/L concentration reduced significantly osteoblast in G0-G1 phase.Of the total groups of Compound Xian Zhen Decoction,osteoblast in S phase increased significantly and 500mg/L concentration showed best function.But in G2-M phase, osteoblast all decreased significantly in Compound Xian Zhen Decoction of 50mg/L and 1mg/L group.CONCLUSIONS:The clinic trail showed Compound Xian Zhen Decoction could partly improve dynamic and static equilibrium in postmenopausal women suffered with osteoporosis. The result was deduced initially to correlate with the improvement of proprioceptive sensibility and muscle power in lower extremity. Therefore the result indicated that Compound Xian Zhen Decoction was benefit for preventing falls.Compound Xian Zhen Decoction amended symptoms of postmenopausal women suffered with osteoporosis,including reducing the pain of lumbar and back.Moreover,it showed better effect than other groups .Compound Xian Zhen Decoction improved BMD of lumbar and retained BMD of neck of femur in postmenopausal women.Compound Xian Zhen Decoction promoted osteoblast-liked to proliferate and 50mg/L concentration performed best effect.Compound Xian Zhen Decoction could inhibit osteoblast apoptosis induced by TNF-a and 1mg/L concentration performed best effect.Different concerntrations of Compound Xian Zhen Decoction effected to different cell cycles.The high concentration effected to G0-G1 and S phases,but lower concerntration effected to G0-G1 , S, and G2-M phases. NEW IDEA:1.The research about Chinese Medicine and movement equilibrium was new at present.The study reviewed a lot of history documents at first,then stated the theory of adjusting dynamic equilibrium in Chinese Medicine and performed clinic trail to research movement equilibrium.The data initially suggested that Chinese herb could influence movement equilibrium by adjusting the balance between Yin and Yang,Zang and Fu,Qi,blood and body fluid in body interior.2.At present,most researches and treatments about osteoporosis were only focused on bone.This study examined the effects of a 6-month Compound Xian Zhen Decoction intervention on muscle strength and balance ability in postmenopausal women .The study researched the contribution of Chinese herb to movement equilibrium of osteoporosis.The data will widen the way of treating osteoporosis.