Sun Guang-Hua, Liao Ying, Liao Yuan, Ni Zhi-Yong, Li Neng, Zhong Pei-Rui, Li Xin-Hong, Zhou Jun
Department of Rehabilitation, the First Affiliated Hospital of University of South China, Hengyang 421001, Hunan Province, China.
Department of Rehabilitation, the First Affiliated Hospital of University of South China, Hengyang 421001, Hunan Province, China; Clinical Medical College of Acupuncture-moxibustion and Rehabilitation, Guangzhou University of Chinese Medicine, Guangzhou 510405.
Zhen Ci Yan Jiu. 2018 Dec 25;43(12):781-7. doi: 10.13702/j.1000-0607.170591.
To observe the effect of electroacupuncture (EA) on histopathological changes of cartilage and subchondral bone and osteoprotegerin (OPG),receptor activator of nuclear factor-κB (RANK), and RANK ligand (RANKL) (OPG/RANK/RANKL) signaling and the expression of matrix metalloproteinase-13 (MMP-13) in ovariectomized(OVX)rats, so as to explore its mechanism underlying improvement of osteoporosis.
Three-month female SD rats were randomly divided into sham operation, model and EA groups (=8 in each group). The ovoariectomy model was established by resection of bilateral ovaries. Rats of the sham group were treated by simple removal of a piece of adipose tissue around the bilateral ovaries. EA (3 Hz/15 Hz, 1 mA) was applied to bilateral "Sanyinjiao" (SP 6), "Yanglingquan" (GB 34), "Yinlingquan" (SP 9) and "Zusanli" (ST 36) for 30 min, once daily (except the weekends) for 12 weeks. The histopathological changes of the subchondral bone of the right knee-joint were observed after Saffron O dyeing and evaluated by Mankin's score, and its anatomical structure including the bone volume fraction (bone volume/tissue volume, BV/TV), trabecular bone number (Tb. N) and trabecular thickness (Tb.Th), trabecular separation (Tb.Sp) was observed by using Micro CT imaging. The urine C-terminal cross-linking telopeptide of type Ⅰ collagen (CTX-Ⅰ), CTX-Ⅱ (two bone resorption markers) and serum estrogen (E 2) contents were assayed by ELISA, and the expression levels of OPG, RANKL and MMP-13 mRNAs in the cartilage tissue of the left knee-joint were detected by quantitative RT-PCR.
Following modeling, the BV/TV, Tb. N and Tb.Th levels were significantly decreased (<0.01) and Tb.Sp and Mankin's score obviously increased in the model group compared with the sham group (<0.01), suggesting a formation of osteoporosis and degeneration of the cartilage tissue. The serum E 2 content and OPG mRNA level in the cartilagetissue were significantly down-regulated (<0.01), and urine CTX-Ⅰ and CTX-Ⅱ contents and RANKL mRNA and MMP-13 mRNA expression levels cartilagetissue were considerably up-regulated in the model group relevant to the sham group (<0.01). After EA intervention, modeling-induced decrease of BV/TV, Tb.N, Tb.Th, E 2 and OPG mRNA levels and OVX-induced increase of Tb.Sp, Mankin's score, CTX-Ⅰ, CTX-Ⅱ, RANKL mRNA and MMP-13 mRNA levels were all completely reversed in the EA group relevant to the model group (<0.01,<0.05)..
EA intervention can inhibit subchondral bone osteoporosis and articular cartilage degeneration of knee-joint in OVX rats, which is closely associated with its effects in inhibiting the down-regulation of serum E 2 and OPG mRNA expression and up-regulation of CTX-Ⅰ, CTX-Ⅱ, RANKL mRNA and MMP-13 mRNA levels, including adjusting OPG/RANK/RANKL signaling.
观察电针(EA)对去卵巢(OVX)大鼠软骨及软骨下骨组织病理学变化、骨保护素(OPG)、核因子κB受体激活剂(RANK)、RANK配体(RANKL)(OPG/RANK/RANKL)信号通路及基质金属蛋白酶-13(MMP-13)表达的影响,以探讨其改善骨质疏松症的作用机制。
将3月龄雌性SD大鼠随机分为假手术组、模型组和电针组(每组8只)。通过切除双侧卵巢建立去卵巢模型。假手术组大鼠仅切除双侧卵巢周围的一片脂肪组织。采用3Hz/15Hz、1mA的电针刺激双侧“三阴交”(SP6)、“阳陵泉”(GB34)、“阴陵泉”(SP9)和“足三里”(ST36),每次30分钟,每日1次(周末除外),共治疗12周。番红花O染色后观察右膝关节软骨下骨的组织病理学变化,采用Mankin评分进行评价,并用Micro CT成像观察其解剖结构,包括骨体积分数(骨体积/组织体积,BV/TV)、骨小梁数量(Tb.N)、骨小梁厚度(Tb.Th)和骨小梁间距(Tb.Sp)。采用酶联免疫吸附测定法(ELISA)检测尿Ⅰ型胶原C端交联肽(CTX-Ⅰ)、CTX-Ⅱ(两种骨吸收标志物)和血清雌激素(E2)含量,采用定量逆转录-聚合酶链反应(RT-PCR)检测左膝关节软骨组织中OPG、RANKL和MMP-13 mRNA的表达水平。
造模后,与假手术组比较,模型组BV/TV水平、Tb.N和Tb.Th水平显著降低(P<0.01),Tb.Sp和Mankin评分明显升高(P<0.01),提示骨质疏松形成及软骨组织退变。与假手术组比较,模型组血清E2含量及软骨组织中OPG mRNA水平显著下调(P<0.01),尿CTX-Ⅰ、CTX-Ⅱ含量及软骨组织中RANKL mRNA和MMP-13 mRNA表达水平显著上调(P<0.01)。电针干预后,与模型组比较,电针组造模引起的BV/TV、Tb.N、Tb.Th、E2和OPG mRNA水平降低及OVX引起的Tb.Sp、Mankin评分、CTX-Ⅰ、CTX-Ⅱ、RANKL mRNA和MMP-13 mRNA水平升高均得到完全逆转(P<0.01,P<0.05)。
电针干预可抑制OVX大鼠膝关节软骨下骨骨质疏松和关节软骨退变,这与其抑制血清E2下调和OPG mRNA表达、上调CTX-Ⅰ、CTX-Ⅱ、RANKL mRNA和MMP-13 mRNA水平的作用密切相关,包括调节OPG/RANK/RANKL信号通路。