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艾灸“关元”“三阴交”对去卵巢大鼠骨形态、代谢及骨髓间充质干细胞ERα的影响

Effect of moxibustion at “Guanyuan”(CV 4) and “Sanyinjiao”(SP 6) on bone morphology,metabolism and ERα of bone marrow mesenchymal stem cells in the ovariectomized rats
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摘要 目的:观察艾灸'三阴交'和'关元'对去卵巢大鼠骨形态、代谢及骨髓间充质干细胞ERα的影响,探讨艾灸关元、三阴交调控骨代谢的内在机制。方法:将60只SD大鼠随机分为正常组(20只)和卵巢切除组(40只),正常组不予处理,卵巢切除组采用经典去卵巢法建立大鼠骨质疏松模型。卵巢切除术后第13周时,正常组和卵巢切除组各取10只进行模型鉴定。模型鉴定成功后,卵巢切除组随机分为模型组、雌二醇组和艾灸组,每组10只。正常组和模型组予2mL 0.9%氯化钠溶液灌胃,每日1次;雌二醇组予17β-雌二醇100μg/(kg·d)灌胃,每日1次;艾灸组予艾灸'关元''三阴交',每穴15 min,每日1次,均6次为一疗程,疗程间休息1 d,共12个疗程。干预12周后,活体状态下采用双能X线测定全身骨骼的骨密度和骨矿质;处死后,分别采用苏木素-伊红染色法(HE)染色观察股骨骨结构,酶联免疫吸附试验测定血清中雌二醇(E2)、碱性磷酸酶(ALP)的吸光度,荧光实时定量PCR法检测胫骨、股骨髓腔中骨髓间充质干细胞ERαmRNA的表达。结果:与模型组比较,艾灸组及雌二醇组大鼠骨盆、脊柱部位的骨密度增加(P<0.05,P<0.01);与模型组比较,艾灸组和雌二醇组大鼠肋骨、骨盆、脊柱等部位的骨矿质含量有升高趋势,但差异无统计学意义(均P>0.05);与模型组比较,艾灸组大鼠骨小梁更加粗壮,相互连接较多;与模型组比较,雌二醇组和艾灸组血清中E2的浓度均升高明显(均P<0.01),雌二醇组和艾灸组血清中ALP的浓度降低明显(均P<0.001),雌二醇组和艾灸组骨髓间充质干细胞中ERαmRNA的表达升高(均P<0.001)。结论:艾灸'三阴交'和'关元'可有效增加去卵巢大鼠的骨密度和骨强度,其机制可能与提高血清中E2浓度,降低血清中ALP浓度,上调骨髓间充质干细胞中ERαmRNA的表达有关。 Objective To observe the effects of moxibustion at 'Guanyuan'(CV 4) and 'Sanyinjiao'(SP 6) on bone morphology, metabolism and ERα of bone marrow mesenchymal stem cells(MSCs) in the ovariectomized rats and explore the underlying mechanism of moxibustion at Guanyuan(CV 4) and Sanyinjiao(SP 6) on the regulation of bone metabolism.Methods A total of 60 SD rats were randomized into a normal group(20 rats) and an ovariectomy group(40 rats). In the normal group, no any treatment was provided. In the ovariectomy group, the classical ovariectomy was adopted to set up the osteoporosis model. In the 13 th week after ovariectomy, 10 rats were collected from each of the normal group and the ovariectomy group for model identification. After model identification, the ovariectomy group was subdivided into a model group, an estradiol group and a moxibustion group, 10 rats in each one. In the normal group and the model group, the gavage was provided with 2 mL 0.9% sodium chloride solution, once a day. In the estradiol group, the gavage was provided with 17β-estradiol 100 μg/(kg·d), once a day. In the moxibustion group, moxibustion was applied at 'Guanyuan'(CV 4) and 'Sanyinjiao'(SP 6), 15 min at each acupoint, once a day. The 6 treatments were as one course, with 1 day of interval, 12 courses were required. After 12-week intervention, the dual-energy X-ray was adopted to determine the bone mineral density(BMD) and bone mineral content of the whole body in living condition. After sacrificed, HE staining was adopted to observe femur structure, the enzyme-linked immunosorbent assay was to determine the absorbance of estradiol(E2) and alkaline phosphatase(ALP) in serum and the real-time quantitative PCR method was to determine the mRNA expressions of ERα of MSCs in tibia and femur. Results Compared with the model group, BMD was increased obviously in the pelvis and spine in the moxibustion group and the estradiol group(P<0.05, P<0.01). Compared with the model group, the bone mineral content was higher in the rib, pelvis and spine
作者 姚长风 胡吴斌 胡玲 王亚玲 王宇 林先刚 刘自兵 YAO Chang-feng;HU Wu-bin;HU Ling;WANG Ya-ling;WANG Yu;LIN Xian-gang;LIU Zi-bing(School of Acupuncture-Moxibustion and Tuina,Anhui University of CM,Hefei 230000,China;the First Affiliated Hospital of Anhui University of CM)
出处 《中国针灸》 CAS CSCD 北大核心 2019年第3期287-292,共6页 Chinese Acupuncture & Moxibustion
基金 安徽省自然科学基金面上项目:1508085MH165 安徽省高校优秀中青年骨干人才海外访学项目:gxfxZD2016123 安徽中医药大学校级教学研究项目:2015xjjy028.
关键词 艾灸 关元 三阴交 去卵巢大鼠 骨密度 moxibustion Point CV 4(Guanyuan) Point SP 6(Sanyinjiao) ovariectomized rats bone mineral density
作者简介 通信作者:胡玲,教授。E-mail:hulingtcm@126.com。
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