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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:

第24卷

期數(shù):

2012年09期

頁碼:

11-14

欄目:

基礎(chǔ)研究

出版日期:

2012-09-30

文章信息/Info

Title:

Regulation of mRNA expression by traditional Chinese medicine JIEGU DECOCTION during fracture healing

作者:

張媛¹; 何延輝²; 王炳南³; 古展群³

1.河南省洛陽正骨醫(yī)院,河南 洛陽 471002;
2.廣東省深圳市橫崗人民醫(yī)院, 廣東 深圳 503800;
3.廣州中醫(yī)藥大學(xué),廣東 廣州 510006

Author(s):

ZHANG Yuan^*; HE Yan-hui; WANG Bing-nan; GU Zhan-qun.^*

Luoyang Orthopedic-Traumatological Hospital,Luoyang 471002,Henan,China

關(guān)鍵詞:

骨折愈合 骨結(jié)合素 接骨方 基因表達(dá)調(diào)控 RNA; 信使 動(dòng)物實(shí)驗(yàn)

Keywords:

Fracture healing;  Osteonectin;  JIEGU DECOCTION;  Gene expression regulation;  RNA; messenger;  Animal experimentation

摘要:

目的:研究中藥接骨方在骨折愈合過程中對(duì)骨粘連蛋白信使核糖核酸表達(dá)的影響,探討中藥接骨方促進(jìn)骨折愈合的作用機(jī)制。方法:將40只2月齡清潔級(jí)雌性SD大鼠制成右股骨中段骨折模型,并行髓內(nèi)固定。造模成功后將40只大鼠分為治療組和對(duì)照組,每組20只。自造模后第1天開始,治療組以接骨方藥液按10.4 mL·kg^-1體質(zhì)量灌胃,對(duì)照組以等體積生理鹽水灌胃。造模后第14天和第28天分別從2組各取10只大鼠處死,取出骨痂組織。將每個(gè)標(biāo)本的骨痂組織按需要分成2份,1份迅速冰凍切片進(jìn)行原位雜交處理,測(cè)定骨粘連蛋白信使核糖核酸表達(dá)情況; 另1份制成石蠟切片,染色后觀察切片中成骨細(xì)胞、破骨細(xì)胞、成軟骨細(xì)胞的情況。結(jié)果:①細(xì)胞學(xué)觀察。造模后第14天,治療組和對(duì)照組均可見較多骨痂形成。治療組骨痂組織中有大量新生骨小梁和軟骨組織,并連接成片,其間成骨細(xì)胞、成軟骨細(xì)胞較為活躍,有少量破骨細(xì)胞; 對(duì)照組纖維組織增生活躍,有大量成骨細(xì)胞形成,并見有一定的軟骨組織。治療組成骨細(xì)胞及破骨細(xì)胞數(shù)量多于對(duì)照組(t=6.520,P=0.000; t=3.670,P=0.002),但成軟骨細(xì)胞數(shù)量少于對(duì)照組(t=12.490,P=0.000)。造模后第28天,治療組骨痂組織中骨小梁數(shù)量較多,粗細(xì)均勻,排列方向一致,相互連接,折光性強(qiáng)。可見由成骨細(xì)胞轉(zhuǎn)化而成并被鈣化骨基質(zhì)包埋的骨細(xì)胞,細(xì)胞形態(tài)正常,有骨小管與基質(zhì)相連。大量排列較為整齊的成骨細(xì)胞和骨細(xì)胞出現(xiàn),破骨細(xì)胞數(shù)量較少。對(duì)照組骨痂中骨小梁排列紊亂,折光強(qiáng)弱不一,并可見小吸收腔,骨折處可見較多纖維細(xì)胞、成軟骨細(xì)胞、成骨細(xì)胞和破骨細(xì)胞。治療組成骨細(xì)胞、成軟骨細(xì)胞及破骨細(xì)胞數(shù)量均少于對(duì)照組(t=6.640,P=0.000; t=9.940,P=0.000; t=8.330,P=0.000)。②骨粘連蛋白信使核糖核酸表達(dá)情況。造模后第14天,治療組大鼠骨痂骨粘連蛋白信使核糖核酸的表達(dá)水平高于對(duì)照組(t=6.700,P=0.000); 造模后第28天,2組大鼠骨痂組織骨粘連蛋白信使核糖核酸表達(dá)水平比較,差異無統(tǒng)計(jì)學(xué)意義(t=1.830,P=0.084)。治療組造模后第28天骨粘連蛋白信使核糖核酸的表達(dá)水平低于造模后第14天(t=4.330,P=0.000); 對(duì)照組造模后第28天骨粘連蛋白信使核糖核酸的表達(dá)水平高于造模后第14天(t=4.770,P=0.000)。結(jié)論:中藥接骨方通過提高骨粘連蛋白信使核糖核酸的表達(dá)水平,使軟骨修復(fù)提早進(jìn)入骨化及塑形期,從而加速骨折愈合。

Abstract:

Objective:To study on the effect of traditional Chinese medicine(TCM)JIEGU DECOCTION on the expression of osteonectin(ON)mRNA during fracture healing,and to explore the mechanism of action of TCM JIEGU DECOCTION in promoting fracture healing.Methods:Forty clean femal SD rats of 2 months old were built models of right mid-femur fractures with intramedullary fixation.After successful molding,40 rats were divided into treatment group and control group,20 cases in each group.Since the first day from molding,rats in the treatment group were intragastric administrated with JIEGU DECOCTION(10.4 mL·kg^-1),while the others in the control group were intragastric administrated with equal volume of normal saline.On the 14th day and 28th day after molding,10 rats selected from each group were executed,and their bony callus tissues were fetched out. Bony callus tissue of each specimen was divided into 2 parts.One part was made into frozen section and processed with in-situ hybridization,then ON mRNA expression was evaluated,while another part was made into paraffin section,and the morphology and amount of osteoblast,osteoclast and chondroblast in the paraffin sections were observed after staining.Results:On the 14th day after molding,massive bony callus tissues were found in treatment group and control group.In the bony callus tissue of treatment group,there were a large amount of new bone trabecula and cartilaginous tissue that connected into pieces,among which the osteoblast and chondroblast were active relatively while only a few number of osteoclasts were found.In the control group,the proliferation of fibrous tissue was active,and massive osteoblasts and a certain number of cartilaginous tissues were found.The amount of osteoblast and osteoclast in the treatment group was larger than that of control group(t=6.520,P=0.000; t=3.670,P=0.002),but the amount of chondroblast in the treatment group was smaller than that of control group(t=12.490,P=0.000).On the 28th day after molding,a large amount of of bone trabeculas were found in the bony callus tissue in treatment group,and they were uniform in thickness and arranged in consistent direction and interconnected with strong light refraction.It was showen that osteocytes transformed from osteoblasts were embedded by calcified bone matrix,the shape of cells were normal,and there were connections between bone canalicules and matrix.Massive osteoblasts and osteocytes appeared and arranged in regular order,while a few amount of osteoclasts were shown.In the control group,bone trabeculas in the bony callus were in a disordered arrangement with uniform strength in refraction,and small absorption cavity was found.Massive fibroblasts,chondroblast,osteoblast and osteoclast were found in the broken ends of fractured bone.The amount of osteoblast,chondroblast and osteoclast in the treatment group were all smaller than those of control group(t=6.640,P=0.000; t=9.940,P=0.000; t=8.330,P=0.000).On the 14th day after molding,ON mRNA expression level in bony callus tissue of rats in the treatment group was higher than that of control group(t=6.700,P=0.000).On the 28th day after molding,there was no statistical difference in ON mRNA expression level in bony callus tissue of rats between the 2 groups(t=1.830,P=0.084).ON mRNA expression level of treatment group on the 28th day after molding was lower than that on the 14th day after molding(t=4.330,P=0.000).ON mRNA expression level in control group on the 28th day after molding was higher than that on the 14th day after molding(t=4.770,P=0.000).Conclusion:JIEGU DECOCTION can raise ON mRNA expression level.As a result,cartilage repair can enter into ossification and remodeling period earlier and the fracture healing is promoted.

備注/Memo

備注/Memo:

2012-02-22收稿 2012-06-14修回

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更新日期/Last Update: 2012-09-30