不同疗程热敏灸干预骨质疏松症模型大鼠穴位周围结缔组织及脊髓的形态学变化

Effect of different courses of heat-sensitive moxibustion on the morphology of surrounding connective tissue and spinal cord in a rat model of osteoporosis

背景:通过临床实践和基础实验证明针灸“命门”穴对治疗原发性骨质疏松症疗效显著,热敏灸作为一种新型治疗手段,在临床治疗中收效甚益,但对其机制的研究仍处于空白阶段。目的:拟从结缔组织及神经系统角度对热敏灸治疗机制进行研究,并在一定时间范围内探讨热敏灸的最佳治疗时间。方法:将SPF级雌性SD大鼠随机分为5组:正常组、模型组、热敏灸4 d组、热敏灸7 d组、热敏灸14 d组,后4组通过双侧卵巢摘除术制备骨质疏松症模型,给予不同时间热敏灸治疗,治疗结束后取以“命门”穴为中心区附近0.5-1.5 cm内的皮下浅筋膜、脊髓分别进行荧光染色、苏木精-伊红染色;对热敏灸14 d治疗前后的血清磷、钙离子和碱性磷酸酶浓度进行测定。结果与结论:①荧光染色观察筋膜内纤维形态改变:模型组的胶原纤维排布交错杂乱呈网状排列,而热敏灸4 d组胶原纤维排列几乎没有任何改变,随着热敏灸时间延长至14 d变化逐渐显著,胶原纤维在热敏灸作用下被拉伸牵引,张力增加,总体呈向心性排布,即朝向“命门”穴附近;②苏木精-伊红染色观察脊髓改变:热敏灸4 d组大鼠脊髓较模型组无明显变化,热敏灸7 d组前角处神经元细胞数量有所增加且体积增大,后角处神经胶质细胞数量增加,热敏灸14 d组前角神经元细胞数目增加显著,且体积增大,细胞核较大而圆,核仁明显,神经纤维增粗增长,排列规律,后角处神经胶质细胞数目明显增加且更加致密;③血清指标:经热敏灸14 d治疗前后对比发现大鼠血清磷离子、碱性磷酸酶浓度明显上升(P<0.01),钙离子浓度明显下降(P<0.05);④结果提示,热敏灸可以通过结缔组织形态学改变以及神经-体液调节来实现热敏信号通路的传导,进而改善骨质疏松的症状,以热敏灸14 d为理想治疗时间。

BACKGROUND:It has been proved in clinical practice and basic research that acupuncture and moxibustion at Mingmen acupoint is an effective treatment for primary osteoporosis.As a new therapeutic method,heat-sensitive moxibustion has achieved good results in clinical practice,but the research on its mechanism is still in the blank stage.OBJECTIVE:To study the therapeutic mechanism of heat-sensitive moxibustion from the perspective of connective tissue and nervous system,and to explore the optimal therapeutic time of heat-sensitive moxibustion in a certain time range.METHODS:Female Sprague-Dawley rats of SPF grade were randomly divided into five groups:normal group,model group,4-,7-,and 14-day heat-sensitive moxibustion groups.Osteoporosis models were prepared by ovariectomy and treated with heat-sensitive moxibustion at different times.After treatment,the superficial subcutaneous fascia 0.5-1.5 cm around the Mingmen acupoint and the spinal cord were stained with fluorescence and hematoxylin-eosin staining.The concentrations of phosphorus ion,calcium ions and alkaline phosphatase in rat serum were detected before and after 14 days of heat-sensitive sensitive moxibustion.RESULTS AND CONCLUSION:(1)Morphological changes of intrafascial fibers by fluorescence staining:In the normal and model groups,collagen fibers were interlaced and arranged in a network.In the 4-day heat-sensitive moxibustion group,the arrangement of collagen fibers showed no changes.After 14 days of intervention the collagen fibers were stretched and pulled significantly under the effect of heat-sensitive moxibustion,and showed a centripetal arrangement toward the Mingmen acupoint.(2)Hematoxylin-eosin staining results of the spinal cord:After 4 days of heat-sensitive moxibustion,the rat spinal cord had no obvious change compared with the model group.After 7 days of heat-sensitive moxibustion,anterior cornual neurons increased in number and size,and the number of glial cells in the spinal dorsal horn increased.After 14 days of heat-sensitive moxibustion,anterior horn neurons increased significantly in number and in size,with large and round nuclei,obvious nucleoli,and enlarged and regularly arranged nerve fibers;glial cells in the posterior horn increased significantly in number and arranged densely.(3)Serum concentration determination:After 14 days of heat-sensitive moxibustion,the concentrations of phosphorus ions and alkaline phosphatase in rat serum increased significantly(P<0.01),and the concentration of calcium ions decreased significantly(P<0.05).These findings indicate that heat-sensitive moxibustion can ameliorate the symptoms of osteoporosis through the morphological changes of connective tissue and nerve-body fluid regulation.Heat-sensitive moxibustion for 14 days can achieve ideal results.