中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2014年
38期
6110-6115
,共6页
马勇%郭杨%屠娟%成吉华%郭各朴%黄桂成
馬勇%郭楊%屠娟%成吉華%郭各樸%黃桂成
마용%곽양%도연%성길화%곽각박%황계성
组织构建%软骨细胞%威灵仙%低强度脉冲超声%Ⅱ型胶原%转化生长因子β1%江苏省自然科学基金
組織構建%軟骨細胞%威靈仙%低彊度脈遲超聲%Ⅱ型膠原%轉化生長因子β1%江囌省自然科學基金
조직구건%연골세포%위령선%저강도맥충초성%Ⅱ형효원%전화생장인자β1%강소성자연과학기금
chondrocytes%Chinese medicine%ultrasound treatment%col agen,type II%tumor necrosis factor-beta1
背景：研究显示威灵仙可促进软骨细胞增殖，维持和促进软骨细胞合成蛋白多糖和Ⅱ型胶原；低强度脉冲超声能有效促进软骨细胞增殖、提高细胞膜的通透性，促进药物或基因的传递，从而提高药物的生物学效应。<br> 目的：观察低强度脉冲超声介导威灵仙对体外培养的兔膝关节软骨细胞增殖、Ⅱ型胶原及转化生长因子β1表达的影响，探讨低强度脉冲超声介导威灵仙在软骨损伤修复中的作用及机制。<br> 方法：体外分离培养兔膝关节软骨细胞，取处于对数生长期的2代细胞，随机分为4组：对照组，低强度脉冲超声组，威灵仙组，威灵仙+低强度脉冲超声组，干预3 d后分别采用CCK-8比色法检测软骨细胞增殖情况，细胞化学染色法分析Ⅱ型胶原分泌情况及Western blot检测转化生长因子β1的表达情况。<br> 结果与结论：干预培养3 d后，与对照组比较，其他3组细胞数量均显著增加(P <0.05)，威灵仙+低强度脉冲超声组细胞数量最多；威灵仙+低强度脉冲超声组软骨细胞Ⅱ型胶原表达面积、吸光度值均显著大于对照组(P <0.05)，且其差值大于低强度脉冲超声组、威灵仙组与对照组差值之和；威灵仙+低强度脉冲超声组转化生长因子β1表达水平显著，其相对灰度值显著高于其他各组(P <0.05)。结果提示低强度脉冲超声、威灵仙均可促进体外培养兔膝关节软骨细胞增殖及Ⅱ型胶原、转化生长因子β1的分泌，二者联用具有一定的协同作用。
揹景：研究顯示威靈仙可促進軟骨細胞增殖，維持和促進軟骨細胞閤成蛋白多糖和Ⅱ型膠原；低彊度脈遲超聲能有效促進軟骨細胞增殖、提高細胞膜的通透性，促進藥物或基因的傳遞，從而提高藥物的生物學效應。<br> 目的：觀察低彊度脈遲超聲介導威靈仙對體外培養的兔膝關節軟骨細胞增殖、Ⅱ型膠原及轉化生長因子β1錶達的影響，探討低彊度脈遲超聲介導威靈仙在軟骨損傷脩複中的作用及機製。<br> 方法：體外分離培養兔膝關節軟骨細胞，取處于對數生長期的2代細胞，隨機分為4組：對照組，低彊度脈遲超聲組，威靈仙組，威靈仙+低彊度脈遲超聲組，榦預3 d後分彆採用CCK-8比色法檢測軟骨細胞增殖情況，細胞化學染色法分析Ⅱ型膠原分泌情況及Western blot檢測轉化生長因子β1的錶達情況。<br> 結果與結論：榦預培養3 d後，與對照組比較，其他3組細胞數量均顯著增加(P <0.05)，威靈仙+低彊度脈遲超聲組細胞數量最多；威靈仙+低彊度脈遲超聲組軟骨細胞Ⅱ型膠原錶達麵積、吸光度值均顯著大于對照組(P <0.05)，且其差值大于低彊度脈遲超聲組、威靈仙組與對照組差值之和；威靈仙+低彊度脈遲超聲組轉化生長因子β1錶達水平顯著，其相對灰度值顯著高于其他各組(P <0.05)。結果提示低彊度脈遲超聲、威靈仙均可促進體外培養兔膝關節軟骨細胞增殖及Ⅱ型膠原、轉化生長因子β1的分泌，二者聯用具有一定的協同作用。
배경：연구현시위령선가촉진연골세포증식，유지화촉진연골세포합성단백다당화Ⅱ형효원；저강도맥충초성능유효촉진연골세포증식、제고세포막적통투성，촉진약물혹기인적전체，종이제고약물적생물학효응。<br> 목적：관찰저강도맥충초성개도위령선대체외배양적토슬관절연골세포증식、Ⅱ형효원급전화생장인자β1표체적영향，탐토저강도맥충초성개도위령선재연골손상수복중적작용급궤제。<br> 방법：체외분리배양토슬관절연골세포，취처우대수생장기적2대세포，수궤분위4조：대조조，저강도맥충초성조，위령선조，위령선+저강도맥충초성조，간예3 d후분별채용CCK-8비색법검측연골세포증식정황，세포화학염색법분석Ⅱ형효원분비정황급Western blot검측전화생장인자β1적표체정황。<br> 결과여결론：간예배양3 d후，여대조조비교，기타3조세포수량균현저증가(P <0.05)，위령선+저강도맥충초성조세포수량최다；위령선+저강도맥충초성조연골세포Ⅱ형효원표체면적、흡광도치균현저대우대조조(P <0.05)，차기차치대우저강도맥충초성조、위령선조여대조조차치지화；위령선+저강도맥충초성조전화생장인자β1표체수평현저，기상대회도치현저고우기타각조(P <0.05)。결과제시저강도맥충초성、위령선균가촉진체외배양토슬관절연골세포증식급Ⅱ형효원、전화생장인자β1적분비，이자련용구유일정적협동작용。
BACKGROUND:Studies have shown that Clematis chinensis Osbeck can promote articular chondrocyte proliferation, maintain and promote the synthesis of cartilage proteoglycan and type II col agen in chondrocytes. Low-intensity pulsed ultrasound can effectively improve the carticular chondrocyte proliferation, increase the cellmembrane permeability, and promote chemicals or genes delivery, so as to improve the biological effects of chemicals. <br> OBJECTIVE:To observe the effect of low-intensity pulsed ultrasound mediated Clematis chinensis Osbeck on the proliferation of cartilage cells and the expression of type II col agen and transforming growth factor (TGF)-β1 of rabbit knee articular chondrocytes cultured in vitro, and explore the effect and mechanism of Clematis chinensis Osbeck mediated by low-intensity pulsed ultrasound on cartilage damage. <br> METHODThe rabbit knee articular chondrocytes were isolated and cultured in vitro. cells of the second generation in logarithmic growth phase were randomly divided into four groupcontrol group, low-intensity pulsed ultrasound (LIPUS) group, Clematis chinensis Osbeck (CCO) group, and CCO+LIPUS group. After cells were cultured under different conditions for 3 days, cellproliferation was measured with cellCount Kit-8 assay. The secretion of type II col agen was detected by cytochemical staining method, and the expression of TGF-β1 was assayed by western blot analysis. <br> RESULTS AND CONCLUSION:The number of cells in the other three groups were significantly higher than that in control group at 3 days after culture (P<0.05), while the CCO+LIPUS group had the most cells. Cytochemical staining showed that the area and absorbance value of type II col agen in cartilage cells were significantly increased in CCO+LIPUS group, compared with control group (P<0.05), and the difference with control group was larger than the amount of that of LIPUS group and CCO group with control group. Western blot analysis showed that TGF-β1 was significantly expressed in CCO+LIPUS group, and the relative gray value was significantly higher than the other groups (P<0.05). Both low-intensity pulsed ultrasound and Clematis chinensis Osbeck can promote the proliferation of cartilage cells and expression of type II col agen and t-β1 of rabbit knee articular chondrocytes cultured in vitro. The combined method could produce synergistic effects.
