| ObjectiveFemale patients with osteoporosis account for the majority of patients with osteoporosis（OP）,and the important reason is the decline of ovarian function,resulting in the decline of estrogen level,resulting in bone loss and bone structure destruction.In recent years,there have been in-depth studies on the study of reverse moxibustion in osteoporosis,but the research on acupoints is not in-depth enough,which brings a lot of inconvenience to clinical application.This study with bone mineral density,bone trabecular structure with hormone levels as the breakthrough point,explores the inverse effect different acupoints reverse moxibustion on to ovarian OP model rats induced by the impact of bone mass and bone structure,looking for inverse moxibustion control key points of the OP and explore the influence of different acupoints for endocrine and bone metabolism,so as to promote,promote acupuncture provide scientific experimental basis for the prevention and treatment of OP.Methods60 clean grade 6-month-old female Sprague Dawley rats were purchased.After 7 days of adaptive feeding,60 rats were randomly numbered,and the last numbers were 1 and 6,2 and 7,3 and 8,4 and 9,5 and 0,respectively,divided into blank group,sham operation group,model group,Weishu group and Shenshu group,with 12 rats in each group.The model group,Weishu group and Shenshu group were used to prepare the osteoporosis model by ovariectomization method.The sham operation group only had partial fat resection on both sides of the ovary,and the other treatments were the same as the model group.Seventy days after the operation,the right femur of rats nos.1-10 and 31-40 was taken and tested by dual-energy X-ray bone density instrument to verify the success of modeling.From the third day of operation,intervention was performed on rats 11~30 and 41~60.The blank group,sham operation group and model group were treated with routine grabbing and self-made rat coat fixation.After skin preparation,reverse moxibustion was suspended at 1~2cm above Weishu point/Shenshu Point in Weishu group and Shenshu group,respectively,for 20min/ day,one course of treatment was performed 6 times,and the rest period was 1 day,a total of 12 courses of treatment.Dual-energy X-ray absorptiometry method were used to detect the femoral bone mineral density（BMD）and HE staining to observe the bone morphology,Micro-CT detection of trabecular bone quantity,connection density index,separating degree,structure model,each to thin,the trabecular bone pattern index factor,structure model,the automatic biochemical analyzer test urine calcium（Ca）,phosphorus（P）urine,urine creatinine（Cr）level,Serum estradiol was detected by ELISA.Results1.Both Shenshu group and Weishu group can improve the bone mineral density of ovariectomy-induced osteoporosis model rats,and the protective effect of Shenshu group is stronger than that of Weishu group.Weishu group and Shenshu group had a tendency to reduce the loss of femur cancellous bone in ovariectomy-induced osteoporosis model rats.（1）Dual-energy X-ray absorption test results: compared with the model group,the BMD of Weishu group and Shenshu group was significantly improved（P<0.05）,and there were also differences between Weishu group and Shenshu group.The BMD of Shenshu group was higher than that of Weishu group（P<0.05）.These results suggest that reverse moxibustion at Shenshu point is stronger than Wei Shu point in preventing bone mineral density reduction in ovariectomized rats with osteoporosis.（2）HE staining results: Comparison of bone cortex: there was no significant difference in the thickness and morphology of bone cortex among all groups.No obvious fracture or gap was found in the internal,external and interplate bone plates.The perforating tube structure was complete and normal in shape,and the overall bone was dense.These results indicate that ovariectomized osteoporosis model rats have little effect on bone.Comparison of Cancellous bone: Compared with the sham operation group,the trabecular fractures in the model group increased,the intermittent distance between trabecular fractures increased,and the trabecular loss appeared in the middle of the model group,showing an osteoporotic trend.Compared with model group,Weishu group and Shenshu group had fewer trabecular fractures,smaller interspace distance between trabecular fractures,trabecular distribution in the middle,and reduced trabecular loss area.These results indicate that reverse moxibustion at Weishu point and Shenshu point can inhibit the loss of bone marrow of femur in ovariectomy-induced osteoporosis model rats.2.Both Weishu group and Shenshu group could prevent and control the bone loss of ovariectomy-induced osteoporosis model rats,and Shenshu group was superior to Weishu group.Both Weishu group and Shenshu group can protect the bone structure of ovariectomy-induced osteoporosis model rats,and Shenshu group is superior to Weishu group.（1）Bone volume fraction results: Compared with the sham operation group,the bone volume fraction of the model group was significantly reduced（P<0.05）,indicating that the trend-induced osteoporosis model rats had serious bone loss.Compared with the model group,the bone volume fraction of Weishu group and Shenshu group was significantly increased（P<0.05）,indicating that reverse moxibustion at Weishu point and Shenshu point could improve the bone mass of ovariectomy-induced osteoporosis model rats.Compared with Weishu group,Shenshu group had higher bone volume and equal number（P<0.05）,indicating that the effect of reverse moxibustion Shenshu on preventing bone loss in ovariectomy-induced osteoporosis model rats was stronger than Weishu group.（2）Bone structure parameters:Compared with the blank group,there was no significant difference in the femur trabecular bone microstructure indexes in the sham group（P>0.05）.These results suggest that the physiological changes caused by surgical trauma alone will not cause significant damage to the bone structure of the subepiphyseal trabecular rich area of femur.Compared with sham operation group,femoral trabecular thickness,trabecular number and trabecular connection density in model group were significantly decreased（P<0.05）,trabecular separation degree,trabecular pattern factor and model structure index were increased（P<0.05）.These results indicate that the changes of estrogen caused by ovariectomization can significantly destroy the bone structure in the rich area of the subepiphyseal trabecular bone of femur.Compared with model group,femoral trabecular thickness,trabecular number and trabecular connection density of Shenshu group were significantly increased（P<0.05,）,trabecular separation degree and trabecular model factor model structure index were decreased（P<0.05）.The thickness of femoral trabecular bone was increased in Weishu group（P<0.05）,but there were no significant differences in bone structure indexes（P>0.05）.These results suggest that reverse moxibustion at Shenshu point can improve the cancellous structure destruction of femur bone in ovariectomy-induced osteoporosis model rats,while Weishu point can only improve the thickness of cancellous trabecular bone in this model rats.Compared with Weishu group,the number and connection density of femoral trabecular bone in Shenshu group were significantly increased（P<0.05,）,and the separation degree of bone trabecular bone and model structure index were decreased（P<0.05）.These results indicate that reverse moxibustion at Shenshu point is superior to Weishu group in improving the cancellous structure of femur bone in ovariectomy-induced osteoporosis model rats.3.Both Shenshu group and Weishu group can reduce the loss of calcium and phosphorus and increase the level of serum estrogen in ovariection-induced osteoporosis model rats,and the effect of Shenshu group is better than that of Weishu group.（1）Results of automatic biochemical apparatus: Compared with the blank group,there were no significant differences in urinary Ca/Cr and URINARY P/Cr in the sham group（P>0.05）,indicating that trauma factors did not affect bone metabolism in rats 3 months after surgical trauma.Compared with blank group and sham group,urine Ca/Cr and URINE P/Cr in model group increased（P<0.05）.Combined with experiment 1 and experiment 2,it showed that ovariectomized rats would lead to abnormal bone metabolism and relatively hyperactivity of bone absorption,leading to osteoporosis.Compared with the model group,urinary Ca/Cr and URINARY P/Cr in Weishu group and Shenshu group were decreased（P<0.05）.Combined with experiment 1 and 2,it was suggested that reverse moxibustion at Weishu and Shenshu 2 points could reduce bone hyperresorption in ovariectomy-induced osteoporosis model rats.Compared with Weishu group,urine Ca/Cr and URINE P/Cr in Shenshu group were decreased（P<0.05）,indicating that Shenshu point was superior to Weishu point in reducing bone hyperresorption in ovariectomy-induced osteoporosis model rats by reverse reverse moxibustion.（2）Elisa assay results: Compared with blank group,the serum estradiol level in sham group had no significant difference（P>0.05）,indicating that surgical trauma did not affect the estradiol level of rats significantly;Compared with blank group and sham operation group,the serum estradiol level of model group was significantly decreased（P<0.05）,indicating that after ovariectomized,estrogen level in rats decreased sharply due to the loss of ovarian secretion.;Compared with model group,the level of serum estradiol in Weishu group and Shenshu group was increased（P<0.05）,suggesting that reverse moxibustion at Weishu and Shenshu points could improve the level of serum estradiol in rats;Compared with Weishu group,serum estradiol level in Shenshu group was increased（P<0.05）,indicating that the level of serum estradiol in Shenshu group was improved by reverse moxibustion,and Shenshu point was better than Weishu point.ConclusionMoxibustion at Weishu point and Shenshu point can increase estrogen level,reduce bone loss and improve bone density,and Shenshu point is better than Weishu point.However,shenshu point has a significant improvement effect on protecting the integrity of bone microstructure,while Weishu point has a weak improvement effect.Therefore,Shenshu point is the key point for the prevention and treatment of OP by moxibustion at Beishu Point.Shenshu point should be selected as the point of moxibustion in clinical and scientific research. |
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