| Objective1Through a randomized clinical study designs, to observe the effect of Bone turnover markers, Clinical pain and Kidney yang deficiency syndrome on postmenopausal osteoporosis. to analysis the prevention of osteoporosis clinical efficacy and mechanism of action.2Through the experimental research to detect the effect of moxibustion serum on osteoporosis osteoblast proliferation and apoptosis, From the cellular and molecular level to investigate its role in the treatment of osteoporosis mechanism.Methods1Clinieal study:A total of90patients with postmenopausal osteoporosis in kidney yang deficiency were assigned, and randomly divided into3groups,30cases in each group, the end of the study does not meet the research requirements and elimination after shedding case, were included in the statistical case87examples, respectively:28cases in the Calcium agent group,30cases in the Proprietary Chinese medicine group and29cases in the mild moxibustion group. Patiets in the Calcium agent group were treated by1Calcium carbonate tablets D3once daily, Patiets in the the Proprietary Chinese medicine group were treated by3Xianlinggubao capsules twice daily and1Calcium carbonate tablets D3once daily. Patiets in the mild moxibustion group were treated by mild moxibustion on the The main points as Pishu point, WeiShu point, ShenShu point and acupoints as Mingmen acupoint acupoints, YaoYangGuan acupoint, ZhiYang acupoint Once every other day and1Calcium carbonate tablets D once daily. All the patients were treated for three months. To Observe the change of Kidney yang deficiency syndrome, Clinical pain, Bone mineral density and Body surface skin temperature of infrared thermograph. According to TCM clinic curative effect evaluation of its curative effect. Meanwhile to determinate patients with bone formation index such as Osteocalcin(BGP), serum bone alkaline phosphatase(BALP), bone resorption index such as tartrate-resistant acid phosphatase5b (TRACP-5b), osteoprotegerin (OPG),bone mineralization indicators serum calcium(Ca), phosphorus(P). Comparison of indexes before and after treatment in group was used with paired sample t test, the differences between groups was used with analysis of variance, LSD, SNK method for statistical analysis, with P<0.05was considered statistically significant criteria.2In vitro experiment:Through the observation of moxibustion serum on rat osteoblast proliferation and apoptosis, on mild-warm moxibustion for treatment of osteoporosis in cell biological mechanism.10born only24hours of neonatal SD rat calvarial bone aseptic were taken. DMEM medium heavy suspension cells came from Using pure collagenase digestion method line of osteoblasts culture, after All culture osteoblasts were inoculated into a plurality of37℃,5%CO2saturated humidity flask uniformed by Wind and percussion. HE staining and ALP staining identification were Conducted, experiments were using of the two generation cell. June old female SD rats were randomly divided into2groups, Each group of10. The rats of moxibustion serum group were treated with gentle moxibustion by moxa cone made by old argy wormwood leaf in rat Back-shu points once a day, each of10minutes for21days, To preparated moxibustion serum and blank serum. The same group of serum after mixing, filtration,56degrees,30minutes bath inactivation aseptic packaging, freezing tube, mark-30℃of cryopreserved standby. Using moxibustion serum added in DMEM medium containing10%, made of moxibustion serum medium, also take blank control group rats serum made with10%serum controls medium. The culture fluid is divided into the blank control group (10%FBS, DMEM), Serum control group (including10%blank serum, DMEM) and moxibustion serum group (including10%moxibustion serum, DMEM) in three groups were cultured respectively, in24hours,48hours after the cultivation,Use MTT method to detect the activity of osteoblast proliferation. According to different culture conditions were divided into control group(DMEM,10%FBS), Apoptosis induced by group (DMEM,10%FBS,40ng/ml TNF alpha), The control group, serum (DMEM,10%blank serum,40ng/ml TNF alpha)and moxibustion serum group (DMEM,10%,40ng/ml TNF moxibustion serum alpha) in four groups, continue to the two generation in cell culture, culture at48hours after completion, do DAPI fluorescence staining, Through an inverted fluorescence microscope osteoblast apoptosis.Result1In clinical studies have shown as follows:Kidney yang deficiency syndrome integral determination results:Calcium group of kidney-yang deficiency syndrome total no difference (P=0.676>0.05), Chinese medicine group of aversion to cold syndrome scores showed no significant difference(P=0.083>0.05), More than indicators have obvious differences (P<0.01), with nocturia and fatigue without evidence of climate change (P=0.001and P=0.003), Mild moxibustion group with aversion to cold, thin integral stool pond is reduced to significantly(P=0.003andP=0.001), P<0.05points more than syndrome. After treatment between group variance analysis comparison of two two, Chinese medicine group, mild moxibustion group and calcium in low back pain and kidney yang deficiency total integral improvement have obvious difference(P<0.05), Mild moxibustion and medicine group comparison only aversion to cold syndrome indices are significantly different(P=0.002<0.01), Other syndromes indices compared with no significant difference (P>0.05). Clinical result of measurement of VAS:Comparison before and after treatment, calcium group for low back pain patients with subjective symptoms do not improve, VAS integral has no obvious change, and Chinese patent medicine group and mild moxibustion group were improved significantly(P=0<0.01). Between group after treatment between the three groups, calcium group VAS integral small changes, Chinese medicine group and mild moxibustion group compared with the VAS integral, a larger decrease(P=0.015<0.05).Infrared thermography of skin temperature measurement results:The results showed, each group before treatment comparison of local temperature had no significant differences within the group, after treatment, mild moxibustion group of local temperature increased significantly (P=0.001<0.01), the other two groups had no significant difference. Comparison between groups, mild moxibustion group of patients than in the other two groups of patients with localized temperature increases, with significant difference (calcium group P=001, P=0.014compared with traditional Chinese Medicine), calcium group and the Chinese medicine group had no significant difference(P=0.389>0.05). Clinical effect of Chinese medicine determination results:Clinical effect of mild moxibustion group accounted for31%, the total efficiency of86.2%. Chinese patent medicine group was33%, the total efficiency of86.7%, Mild moxibustion group and the traditional Chinese medicine curative effect had no significant difference, But were significantly better than the calcium group (P=0.007<0.01).Determination of bone mineral density BMD results:the group after treatment, There were no significant differences.Bone turnover markers determination results:The three groups before treatment comparison, each index showed no significant differences, with comparable (P>0.05), BGP group, Chinese medicine group and mild moxibustion group were significantly different(P=0<0.01), calcium group had no significant difference. BGP comparison between groups, mild moxibustion group than the calcium group increased significantly(P=0.004<0.01),a proprietary Chinese medicine group was significantly (P=0.075>0.05). BALP group, calcium groups before and after treatment was not significantly different,proprietary Chinese medicine group BALP increased (P=0.001<0.01), Mild moxibustion increased amplitude slightly larger (P=0<0.01).Between group and calcium compared, there are some differences(P<0.01). Chinese medicine and moxibustion group significantly increased compared to the value have no significant difference,both in the normal range(P=0.306).Three groups of TRACP5b decreased calcium groups before and after treatment, no differences, Chinese medicine and moxibustion group before and after treatment are compared, have significant difference (P<0.01), the three group after treatment, calcium group and there are significant differences between Chinese medicine group(P=0.005<0.01), and mild moxibustion group comparison difference(P=0.013<0.05), Chinese medicine group and mild moxibustion groups have not obvious difference. OPG group compared esultst:there were significant differences of proprietary Chinese medicine group, mild moxibustion has significant difference. But after treatment were compared between groups, mild moxibustion group and calcium group, have a certain degree of down(P=0.044<0.05), Chinese medicine group and calcium group differences(P=0.046<0.05).The three groups before and after treatment and compared between groups, serum calcium, phosphorus levels have no significant difference.2Experimental studies have shown, the blank control component of bone cell apoptosis rate and cell apoptosis of mean lower than the other three groups, moxibustion serum composition of bone cell apoptosis, apoptosis and apoptosis induced by serum mean group and the comparison group, there are significant differences (P<0.05), serum controls and apoptosis induced by group comparison no significant difference(P>0.05). Three groups in the24hours of osteoblast proliferation od no difference (P>0.05), At48hours after each OD570obviously increased, The proliferation of moxibustion serum group greatly, and the blank control group, and serum in comparison to the control group, there were significant differences (P<0.05), Serum control group compared with the control group no significant difference(P>0.05). In different periods, the blank control group and control group the OD value of serum were increased, but there is no obvious difference in comparison within groups (P>0.05), moxibustion serum groups in different time slots within group comparisons, osteoblast OD570value were significantly increased.Conclusion1Mild moxibustion can obviously improve the kidney yang deficiency in postmenopausal osteoporosis patients clinical symptoms, relief of pain in osteoporosis, and is superior to other treatment group.2Mild moxibustion can improve of osteoporosis with lumbar pain around the site and the infrared skin temperature value, effectively improve the kidney yang deficiency syndrome.3Mild moxibustion can improve bone formation related indexes, decreased bone resorption related indicators, and bone turnover the kidney yang deficiency in postmenopausal osteoporosis patients.4Moxibustion serum can promote the proliferation of osteoblasts, can reduce the TNF alpha induced apoptosis of osteoblastic cells number. |
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