独活寄生汤对骨性关节炎软骨细胞增殖与凋亡影响的实验研究
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摘要
目的
探讨独活寄生汤对骨性关节炎大鼠关节软骨细胞增殖与凋亡的影响。
方法
1.健康2月龄清洁级SD大鼠144只,雌雄各半,适应性喂养1周,随机分为空白组(36只)和造模组(108只)。造模组在1,4,7天双膝关节腔注射4%木瓜蛋白酶复制骨性关节炎模型。造模成功后,抽签法随机分为模型组(36只)、治疗组(36只)和阳性组(36只)。空白组和模型组按照10ml·kg-1·d-1的量灌服0.9%生理盐水；治疗组按照9.3g.kg-1·d-1的药量灌服独活寄生汤；阳性组按照0.15g-kg-1d-1的药量灌服奥泰灵胶囊；每2周为1个疗程,中间休息2天,共4个疗程。
2.每2个疗程后,处死一批实验动物,切取胫骨平台内侧软骨组织于4%多聚甲醛中固定、石蜡包埋,HE染色观察软骨组织形态变化；免疫组化检测Cyclin D1、CDK4、 Rb、p16、Bc1-2、Bax、Cytochrome c、caspase-9及caspase-3蛋白表达情况；切取胫骨平台外侧软骨组织于电镜固定液中固定,用于观察软骨细胞超微结构变化；其余软骨组织迅速置入液氮中保存,用于检测Cyclin D1、CDK4、Rb及p16mRNA表达情况。
3.健康2月龄清洁级SD大鼠144只,雌雄各半,抽签法随机分为：不用药血清对照组(空白血清组,36只)和独活寄生汤临床等效剂量组(含药血清组,108只)。空白血清组按10ml·kg-1·d-1的量给予0.9%生理盐水；含药血清组按9.3g-kg-1-d-1的量给予独活寄生汤。连续7天灌胃,最后1天连续2次灌胃,中间间隔2h,采血前禁食12h,分别在末次灌胃1h、2h、3h后,在2%戊巴比妥钠2ml/kg麻醉下腹主动脉采血,制备含药血清。
4.健康4周龄清洁级SD大鼠20～30只,取膝关节的关节软骨,建立软骨细胞体外培养体系。第2代软骨细胞以1.0×104/孔接种于96孔板中,在含10%FBS的DMEM培养基中培养24h后,用不含FBS的DMEM培养液培养24h,同步化软骨细胞。分别加入含10%、15%、20%、30%四个浓度的不同采血时间点的含药血清和空白血清,培养24h、36h、48h、60h、72h后,MTT法检测软骨细胞的活性。
5.用10ng/ml IL-1β诱导第3代软骨细胞复制退变软骨细胞模型,采用倒置相差显微镜观察细胞形态变化和Ⅱ型胶原免疫组化进行鉴定。
6.采用(6)中确定的含药血清最佳干预条件,体外培养退变软骨细胞,分别干预24h,48h,72h后,收集软骨细胞。MTT法检测细胞增殖情况；流式细胞术检测细胞周期变化情况；Annexin V-FITC/PI染色流式检测细胞凋亡情况；JC-1染色流式检测含药血清干预后线粒体膜电位的改变情况; RT-PCR法检测Cyclin D1、CDK4、Rb、p16、Bcl-2、 Bax mRNA表达情况；Western Blot法检测Cyclin D1、CDK4、Rb、p16、Bcl-2、Bax蛋白表达情况；分光光度法检测caspase-9和caspase-3活化情况。
结果
1.膝关节X侧位片示,正常组关节间隙正常、关节面平整、关节边缘可见光滑锐利的线样致密影,软骨下骨密度均匀。模型组关节间隙明显变窄、关节面不平整,软骨破坏区边缘骨质增生硬化,关节边缘骨赘形成。HE染色和电镜观察结果表明独活寄生汤和奥泰灵均可以促进OA软骨的修复,促进软骨细胞增殖,两者无明显差别。独活寄生汤和奥泰灵均能促进Cyclin D1、CDK4、Rb mRNA和蛋白,抑制p16、Cytochrome c、 caspase-9及caspase-3mRNA和蛋白的表达,两者无显著性差异。
2.第3代软骨细胞Ⅱ型胶原免疫组织化学染色,软骨细胞胞浆被染成棕黄色,胞核不着色；阿利新蓝染色,软骨细胞核为浅蓝色,细胞质中可见不少浅蓝色的分泌颗粒和小泡。独活寄生汤含药血清干预软骨细胞增殖最佳采血时间点为2小时,最佳干预浓度为10%。采用10ng/ml IL-1β诱导的软骨细胞,与正常细胞相比,细胞体积变大,边缘出现一些指状突起,核也变大,胞膜和胞浆不清晰,细胞呈多边形改变,折光度下降；Ⅱ型胶原免疫组化显示软骨细胞胞浆棕黄色染色变浅。含药血清干预后,退变软骨细胞G0/G1期比例逐渐降低,S期比例和增殖指数逐渐增加；退变软骨细胞的凋亡率逐渐降低；线粒体膜电位逐渐降低；随着干预时间的延长,caspase-9和caspase-3活性均逐渐降低；含药血清能促进Cyclin D1、CDK4、Rb、Bcl-2mRNA和蛋白表达,抑制p16和Bax mRNA和蛋白的表达。
结论
1.体内外研究表明,独活寄生汤通过促进软骨细胞周期关键限制点G1/S期的切换,促进软骨细胞增殖。
2.体内外研究表明,独活寄生汤通过抑制线粒体凋亡通路的激活,抑制软骨细胞凋亡。
Objective
To discuss the effect of Duhuo Jisheng Decoction (DHJSD) on osteoarthritis chondrocyte proliferation and apoptosis.
Methods
1. After one week of acclimation, healthy and clean, two-month-old Sprague Dawley rats of average gender (n=144) were randomly divided into blank group (n=36) and osteoarthritis model group (n=108). Osteoarthritis model was induced in double knees of rats by injecting0.2ml of4%papain solution on days1,4and7. After successfully establish the model, the animals were randomly divided into three groups:the model group (received an equivalent amount of saline only), the positive control group [received a clinical oral dose of Glucosamine Hydrochloride Capsules (0.15g/kg/day)] and the treatment group [received a clinical oral dose of DHJSD (9.3g/kg/day)]. The blank group also received an equivalent amount of saline only. Every treatment course includes two weeks. There are totally four courses.
2. After every two courses, the animals were sacrificed and tibial plateau cartilage specimens were obtained. The morphological changes were observed under an optical microscope following staining with hematoxylin and eosin (H&E) and by transmission electron microscopy (TEM). The protein levels of Cyclin D1、CDK4、Rb、p16、Bcl-2、Bax、 Cytochrome c、Caspase-9and caspase-3were measured by immunohistochemistry. The mRNA levels of Cyclin D1、CDK4、 Rb and p16were measured by RT-PCR.
3. A total of144two-month-old Sprague Dawley rats of average gender were randomly divided into two groups:the DHJSD group (n=108) treated with a dose of9.3g/kg/day DHJSD, which is the equivalent dosage for human in clinical use and the blank group (n=36) treated with an equivalent amount of saline only. All testing drugs and saline were administered via gastric gavage twice a day in the morning and afternoon for7consecutive days. The daily doses were given2h apart on the seventh day. Animals were anesthetized by intraperitoneal injection of2ml/kg2%pentobarbital sodium, and arterial blood in DHJSD group was collected from the abdominal aorta at1h,2h and3h after the final dose in DHJSD group, respectively and at2h in blank group.
4. Cartilage was isolated from the knee joint of20～30health and clean, four-week-old SD rats, and used to establish a cultivation system of chondrocytes in vitro. The second-generation chondrocytes were seeded into96-well plates at a density of1.0x105cells/ml in0.1ml of medium. After synehronization, the cells were treated with10%、15%、20%、30%concentrations of different sampling time of DHJSD serum for24,36,48,60and72h, respectively. Cell viability was assessed by the MTT colorimetric assay.
5. The degenerative chondrocyte model was established by exposing to10ng/ml IL-1β for24h, and verified by optical microscopy analyses and immunohistochemical staining of Collagen II analyses.
6. After treated degenerative chondrocytes with the best intervention condition for24h,48h and72h, cell proliferation was detected by MTT assay and DNA staining followed by FACS analysis; cell apoptosis was detected by Flow Cytometry Analysis with Annexin V-FITC/PI Staining; mitochonrial membrane potential (ΔΨm) was measured by Flow Cytometry analysis with JC-1staining; Caspase-9,-3activation was analysesed by spectrophotometry; and the mRNA and protein expression levels of Cyclin D1, CDK4, Rb, p16, Bcl-2and Bax were measured by RT-PCR and Western Blot, respectively.
Results
1. The knee lateral X ray showed normal joint space, smooth articular surface, smooth lines dense shadow on joint edge and uniformity bone density in subchondral in normal group, while in model group, the joint space became narrow; the articular surface was not flat; on the edge of damage cartilage appeared bone hyperplasia and sclerosis; joint marginal appeared osteophyte formation. HE staining and transmission electron microscopy (TEM) results showed DHJSD and Glucosamine Hydrochloride Capsules could both promote OA cartilage repair and chondrocytes proliferation with no significant difference. DHJSD and Glucosamine Hydrochloride Capsules could both promote the mRNA and protein expression of Cyclin Dl, CDK4, Rb and inhibit p16, Cytochrome c, caspase-9, caspase-3mRNA and protein expression with no significant difference.
2. Identification of the third generation chondrocytes with immunohistochemical staining of Collagen Ⅱ, the cytoplasm appeared brown and no coloration in the nucleus; with alcian blue staining, the nucleus were stained light blue, a lot of secretion particles and vesicles appeared in the cytoplasm. The best sampling time and concentration of DHJSD serum on promoting chondrocyte proliferation were2hour and10%, respectively. Compared with the normal chondrocytes,10ng/ml IL-1β induced chondrocytes became larger with some finger-like protrusions on the edge, and the cell membrane and cytoplasm was not clear. Meanwhile, the IL-1β-induced cells were polygonal changed, along with declining refraction. Besides, the immunohistochemical staining of Collagen II analyses showed that the brownish yellow staining in cytoplasm became shallow. After treated with DHJSD2h serum, the percentage of G0/G1phase cells was gradually decreased; however, the percentage of S phase cells and the proliferation index were increased. DHJSD2h serum treatment inhibited the apoptosis of degenerative chondrocytes, the loss of plasma membrane asymmetry (externalization of phosphatidylserine), collapse of mitochondrial membrane potential, and activation of caspase-9and caspase-3. DHJSD serum could promote the mRNA and protein expression of Cyclin D1, CDK4, Rb, Bcl-2and inhibit the mRNA and protein expression of p16, Bax.
Conclusion
1. DHJSD could promote osteoarthritis chondrocytes proliferation by promoting G1/S transition, in vivo and vitro.
2. DHJSD could inhibit the mitochondrion-dependent pathway of apoptosis in osteoarthritis chondrocytes, in vivo and vitro.
探讨独活寄生汤对骨性关节炎大鼠关节软骨细胞增殖与凋亡的影响。
方法
1.健康2月龄清洁级SD大鼠144只,雌雄各半,适应性喂养1周,随机分为空白组(36只)和造模组(108只)。造模组在1,4,7天双膝关节腔注射4%木瓜蛋白酶复制骨性关节炎模型。造模成功后,抽签法随机分为模型组(36只)、治疗组(36只)和阳性组(36只)。空白组和模型组按照10ml·kg-1·d-1的量灌服0.9%生理盐水；治疗组按照9.3g.kg-1·d-1的药量灌服独活寄生汤；阳性组按照0.15g-kg-1d-1的药量灌服奥泰灵胶囊；每2周为1个疗程,中间休息2天,共4个疗程。
2.每2个疗程后,处死一批实验动物,切取胫骨平台内侧软骨组织于4%多聚甲醛中固定、石蜡包埋,HE染色观察软骨组织形态变化；免疫组化检测Cyclin D1、CDK4、 Rb、p16、Bc1-2、Bax、Cytochrome c、caspase-9及caspase-3蛋白表达情况；切取胫骨平台外侧软骨组织于电镜固定液中固定,用于观察软骨细胞超微结构变化；其余软骨组织迅速置入液氮中保存,用于检测Cyclin D1、CDK4、Rb及p16mRNA表达情况。
3.健康2月龄清洁级SD大鼠144只,雌雄各半,抽签法随机分为：不用药血清对照组(空白血清组,36只)和独活寄生汤临床等效剂量组(含药血清组,108只)。空白血清组按10ml·kg-1·d-1的量给予0.9%生理盐水；含药血清组按9.3g-kg-1-d-1的量给予独活寄生汤。连续7天灌胃,最后1天连续2次灌胃,中间间隔2h,采血前禁食12h,分别在末次灌胃1h、2h、3h后,在2%戊巴比妥钠2ml/kg麻醉下腹主动脉采血,制备含药血清。
4.健康4周龄清洁级SD大鼠20～30只,取膝关节的关节软骨,建立软骨细胞体外培养体系。第2代软骨细胞以1.0×104/孔接种于96孔板中,在含10%FBS的DMEM培养基中培养24h后,用不含FBS的DMEM培养液培养24h,同步化软骨细胞。分别加入含10%、15%、20%、30%四个浓度的不同采血时间点的含药血清和空白血清,培养24h、36h、48h、60h、72h后,MTT法检测软骨细胞的活性。
5.用10ng/ml IL-1β诱导第3代软骨细胞复制退变软骨细胞模型,采用倒置相差显微镜观察细胞形态变化和Ⅱ型胶原免疫组化进行鉴定。
6.采用(6)中确定的含药血清最佳干预条件,体外培养退变软骨细胞,分别干预24h,48h,72h后,收集软骨细胞。MTT法检测细胞增殖情况；流式细胞术检测细胞周期变化情况；Annexin V-FITC/PI染色流式检测细胞凋亡情况；JC-1染色流式检测含药血清干预后线粒体膜电位的改变情况; RT-PCR法检测Cyclin D1、CDK4、Rb、p16、Bcl-2、 Bax mRNA表达情况；Western Blot法检测Cyclin D1、CDK4、Rb、p16、Bcl-2、Bax蛋白表达情况；分光光度法检测caspase-9和caspase-3活化情况。
结果
1.膝关节X侧位片示,正常组关节间隙正常、关节面平整、关节边缘可见光滑锐利的线样致密影,软骨下骨密度均匀。模型组关节间隙明显变窄、关节面不平整,软骨破坏区边缘骨质增生硬化,关节边缘骨赘形成。HE染色和电镜观察结果表明独活寄生汤和奥泰灵均可以促进OA软骨的修复,促进软骨细胞增殖,两者无明显差别。独活寄生汤和奥泰灵均能促进Cyclin D1、CDK4、Rb mRNA和蛋白,抑制p16、Cytochrome c、 caspase-9及caspase-3mRNA和蛋白的表达,两者无显著性差异。
2.第3代软骨细胞Ⅱ型胶原免疫组织化学染色,软骨细胞胞浆被染成棕黄色,胞核不着色；阿利新蓝染色,软骨细胞核为浅蓝色,细胞质中可见不少浅蓝色的分泌颗粒和小泡。独活寄生汤含药血清干预软骨细胞增殖最佳采血时间点为2小时,最佳干预浓度为10%。采用10ng/ml IL-1β诱导的软骨细胞,与正常细胞相比,细胞体积变大,边缘出现一些指状突起,核也变大,胞膜和胞浆不清晰,细胞呈多边形改变,折光度下降；Ⅱ型胶原免疫组化显示软骨细胞胞浆棕黄色染色变浅。含药血清干预后,退变软骨细胞G0/G1期比例逐渐降低,S期比例和增殖指数逐渐增加；退变软骨细胞的凋亡率逐渐降低；线粒体膜电位逐渐降低；随着干预时间的延长,caspase-9和caspase-3活性均逐渐降低；含药血清能促进Cyclin D1、CDK4、Rb、Bcl-2mRNA和蛋白表达,抑制p16和Bax mRNA和蛋白的表达。
结论
1.体内外研究表明,独活寄生汤通过促进软骨细胞周期关键限制点G1/S期的切换,促进软骨细胞增殖。
2.体内外研究表明,独活寄生汤通过抑制线粒体凋亡通路的激活,抑制软骨细胞凋亡。
Objective
To discuss the effect of Duhuo Jisheng Decoction (DHJSD) on osteoarthritis chondrocyte proliferation and apoptosis.
Methods
1. After one week of acclimation, healthy and clean, two-month-old Sprague Dawley rats of average gender (n=144) were randomly divided into blank group (n=36) and osteoarthritis model group (n=108). Osteoarthritis model was induced in double knees of rats by injecting0.2ml of4%papain solution on days1,4and7. After successfully establish the model, the animals were randomly divided into three groups:the model group (received an equivalent amount of saline only), the positive control group [received a clinical oral dose of Glucosamine Hydrochloride Capsules (0.15g/kg/day)] and the treatment group [received a clinical oral dose of DHJSD (9.3g/kg/day)]. The blank group also received an equivalent amount of saline only. Every treatment course includes two weeks. There are totally four courses.
2. After every two courses, the animals were sacrificed and tibial plateau cartilage specimens were obtained. The morphological changes were observed under an optical microscope following staining with hematoxylin and eosin (H&E) and by transmission electron microscopy (TEM). The protein levels of Cyclin D1、CDK4、Rb、p16、Bcl-2、Bax、 Cytochrome c、Caspase-9and caspase-3were measured by immunohistochemistry. The mRNA levels of Cyclin D1、CDK4、 Rb and p16were measured by RT-PCR.
3. A total of144two-month-old Sprague Dawley rats of average gender were randomly divided into two groups:the DHJSD group (n=108) treated with a dose of9.3g/kg/day DHJSD, which is the equivalent dosage for human in clinical use and the blank group (n=36) treated with an equivalent amount of saline only. All testing drugs and saline were administered via gastric gavage twice a day in the morning and afternoon for7consecutive days. The daily doses were given2h apart on the seventh day. Animals were anesthetized by intraperitoneal injection of2ml/kg2%pentobarbital sodium, and arterial blood in DHJSD group was collected from the abdominal aorta at1h,2h and3h after the final dose in DHJSD group, respectively and at2h in blank group.
4. Cartilage was isolated from the knee joint of20～30health and clean, four-week-old SD rats, and used to establish a cultivation system of chondrocytes in vitro. The second-generation chondrocytes were seeded into96-well plates at a density of1.0x105cells/ml in0.1ml of medium. After synehronization, the cells were treated with10%、15%、20%、30%concentrations of different sampling time of DHJSD serum for24,36,48,60and72h, respectively. Cell viability was assessed by the MTT colorimetric assay.
5. The degenerative chondrocyte model was established by exposing to10ng/ml IL-1β for24h, and verified by optical microscopy analyses and immunohistochemical staining of Collagen II analyses.
6. After treated degenerative chondrocytes with the best intervention condition for24h,48h and72h, cell proliferation was detected by MTT assay and DNA staining followed by FACS analysis; cell apoptosis was detected by Flow Cytometry Analysis with Annexin V-FITC/PI Staining; mitochonrial membrane potential (ΔΨm) was measured by Flow Cytometry analysis with JC-1staining; Caspase-9,-3activation was analysesed by spectrophotometry; and the mRNA and protein expression levels of Cyclin D1, CDK4, Rb, p16, Bcl-2and Bax were measured by RT-PCR and Western Blot, respectively.
Results
1. The knee lateral X ray showed normal joint space, smooth articular surface, smooth lines dense shadow on joint edge and uniformity bone density in subchondral in normal group, while in model group, the joint space became narrow; the articular surface was not flat; on the edge of damage cartilage appeared bone hyperplasia and sclerosis; joint marginal appeared osteophyte formation. HE staining and transmission electron microscopy (TEM) results showed DHJSD and Glucosamine Hydrochloride Capsules could both promote OA cartilage repair and chondrocytes proliferation with no significant difference. DHJSD and Glucosamine Hydrochloride Capsules could both promote the mRNA and protein expression of Cyclin Dl, CDK4, Rb and inhibit p16, Cytochrome c, caspase-9, caspase-3mRNA and protein expression with no significant difference.
2. Identification of the third generation chondrocytes with immunohistochemical staining of Collagen Ⅱ, the cytoplasm appeared brown and no coloration in the nucleus; with alcian blue staining, the nucleus were stained light blue, a lot of secretion particles and vesicles appeared in the cytoplasm. The best sampling time and concentration of DHJSD serum on promoting chondrocyte proliferation were2hour and10%, respectively. Compared with the normal chondrocytes,10ng/ml IL-1β induced chondrocytes became larger with some finger-like protrusions on the edge, and the cell membrane and cytoplasm was not clear. Meanwhile, the IL-1β-induced cells were polygonal changed, along with declining refraction. Besides, the immunohistochemical staining of Collagen II analyses showed that the brownish yellow staining in cytoplasm became shallow. After treated with DHJSD2h serum, the percentage of G0/G1phase cells was gradually decreased; however, the percentage of S phase cells and the proliferation index were increased. DHJSD2h serum treatment inhibited the apoptosis of degenerative chondrocytes, the loss of plasma membrane asymmetry (externalization of phosphatidylserine), collapse of mitochondrial membrane potential, and activation of caspase-9and caspase-3. DHJSD serum could promote the mRNA and protein expression of Cyclin D1, CDK4, Rb, Bcl-2and inhibit the mRNA and protein expression of p16, Bax.
Conclusion
1. DHJSD could promote osteoarthritis chondrocytes proliferation by promoting G1/S transition, in vivo and vitro.
2. DHJSD could inhibit the mitochondrion-dependent pathway of apoptosis in osteoarthritis chondrocytes, in vivo and vitro.
引文
[1].Roush JK, Cross AR, Renberg WC, et al. Evaluation of the effects of dietary supplementation with fish oil omega-3 fatty acids on weight bearing in dogs with osteoarthritis.J Am Vet Med Assoc.2010,236(1):67-73.
[2].李西海,梁文娜.软骨下骨骨重塑与骨关节炎的关系[J].国际骨科学杂志.2007,28(1)：35-37.
[3].Dahm DL, Al-Rayashi W, Dajani K, et al. Patellofemoral arthroplasty versus total knee arthroplasty in patients with isolated patellofemoral osteoarthritis.Am J Orthop (Belle Mead NJ).2010,39(10):487-491.
[4].Vuolteenaho K, Moilanen T, Knowles RG, et al. The role of nitric oxide in osteoarthritis. Scand J Rheumatol.2007,36(4):247-258.
[5].Shakoor MA, Rahman MS, Azad AK, et al. Effects of isometric quadriceps muscle strengthening exercise on chronic osteoarthritis of the knee.Bangladesh Med Res Counc Bull.2010,36(1):20-22.
[6].李宁华,薛庆云,张毅,等.中国六城市中老年人群X线膝骨关节炎流行病学分析[J].实用医学杂志.2008,24(16)：2887-2888.
[7].周青,王宇仁,周之德,等.不同分子量的透明质酸钠治疗膝骨关节炎的比较[J].上海第二医科大学学报.2000,20(4)：361-363.
[8].邱贵兴.骨关节炎流行病学和病因学进展[J].继续医学教育.2005,19(7)：68-69.
[9].周肃陵,刘肖瑜,王正.独活寄生汤在骨伤科的临床应用[J].中医正骨.2006,18(12)：67.
[10].李西海,陈文列,刘献祥.补肾柔肝法防治骨性关节炎作用机制探讨[J].福建中医药大学学报.2011,21(2)：66-69.
[11].郑春松,叶蕻芝,许惠凤,等.分子对接法预测独活寄生汤对INF-α和IL-1β作用的活性成分[J].福建中医药大学学报.2012,22(4)：28-30.
[12].王武炼,叶锦霞,刘献祥,等.独活寄生汤加减内服外洗治疗膝骨性关节炎66例临床观察[J].福建中医药大学学报.2011,21(2)：44-45.
[13].李西海,张翼,叶蕻芝,等.独活寄生汤治疗凤寒湿痹型膝骨性关节炎的机制探讨[J].中医正骨.2012,24(1)：68-71.
[14].吴广文,褚剑锋,许惠风,等.独活寄生汤的药理作用及其在治疗骨性关节炎中的应用[J].中医正骨.2012,24(1)：37-39.
[15].郑春松,叶蕻芝,许惠凤,等.从化学空间角度看独活寄生汤治疗骨性关节炎的多靶点作用[J].福建中医药大学学报.2011,21(5)：35-37.
[16].Zheng CS, X XJ, Y HZ, et al. Computational approaches for exploring the potential synergy and polypharmacology of Duhuo Jisheng Decoction in the therapy of osteoarthritis. Mol Med Report.2013,7(6):1812-1818.
[17].Haq I, Murphy E, Dacre J. Osteoarthritis. Postgrad Med J.2003,79(933):377-383.
[18].Uryu N, Okada K, Kawakita K. Analgesic effects of indirect moxibustion on an experimental rat model od osteoarthritis in the knee.Acupunct Med.2007,25(4):175-183.
[19].Torelli SR, Rahal SC, Volpi RS, et al. Histopathological evaluation of treatment with chondroitin sulphate for osteoarthritis induced by continuous immobilization in rabbits. J Vet Med A Physiol Pathol Clin Med.2005,52(1):45-51.
[20].Legendre F, Heuze A, Boukerrouche K, et al. Rhein, the metabolite of diacerhein, reduces the proliferation of osteoarthritic chondrocytes and synoviocytes without inducing apoptosis.Scand J Rheumatol.2009,38(2):104-111.
[21].Matsumoto T, Cooper GM, Gharaibeh B, et al. Cartilage repair in a rat model of osteoarthritis through intraarticular transplantation of muscle-derived stem cells expressing bone morphogenetic protein 4 and soluble flt-1.Arthritis Rheum.2009,30(5):1390-1405.
[22].Guo D, Ding L, Homandberg GA. Telopeptides of type Ⅱ collagen upregulate proteinases and damage cartilage but are less effective than highly active fibronectin fragments. Inflamm Res.2009,58(3):161-169.
[23].Li X, Ellman M, Muddasani P, et al. Prostaglandin E2 and its cognate EP receptors control human adult articular cartilage homeostasis and are linked to the pathophysiology of osteoarthritis.Arthritis Rheum.2009,30(2):513-523.
[24].Yang X, Chen L, Xu X, et al. TGF-β/Smad3 signals repress chondrocyte hypertrophic differentiation and are required for maintaining articular cartilage.J Cell Biol.2001,153(1):35-46.
[25].Iannone F, De Bari C, Scioscia C, et al. Increased Bcl-2/p53 ratio in human osteoarthritic cartilage:a possible role in regulation of chondrocyte metabolism.Ann Rheum Dis.2005,64(2):217-221.
[26].Zhou HW, Lou SQ, Zhang K. Recovery of function in osteoarthritic chondrocytes induced by p16INK4a-siRNA in vitr. Rheumatology(Oxford).2004,43(5):555-568.
[27].Tetlow LC, Woolley DE. Histamine stimulates the proliferation of human articular chondrocytes in vitro and is expressed by chondrocytes in osteoarthritic cartilage.Ann Rheum Dis.2003,62(10):991-994.
[28].Sharma L, Kapoor D, Issa S. Epidemiology of osteoarthritis:an update.Curr Opin Rheumatol.2006,18(2):147-156.
[29].Sandell LJ, Aigner T. Articular cartilage and changes in arthritis. An introduction:Cell biology of osteoarthritis.Arthritis Res.2001,3(2):107-113.
[30].Malemud CJ. Protein kinases in chondrocyte signaling and osteoarthritis. Clin Orthop Relat Res.2004,(427 Supp 1):145-151.
[31].Mistry D, Oue Y, Chambers MG, et al. Chondrocyte death during murine osteoarthritis[J]. Osteoarthritis Cartilage.2004,12(2):131-141.
[32].Embree MC, Kilts TM, Ono M, et al. Biglycan and fibromodulin have essential roles in regulating chondrogenesis and extracellular matrix turnover in temporomandibular joint osteoarthritis.Am J Pathol.2010,176(2):812-826.
[33].Kunitomo T, Takahashi KA, Arai Y, et al. Influence of extracellular matrix on the expression of inflammatory cytokines, proteases, and apoptosis-related genes induced by hydrostatic pressure in three-dimensionally cultured chondrocytes.J Orthop Sci.2009,14(6):776-783.
[34].Giovannini S, Diaz-Romero J, Aigner T, et al. Population doublings and percentage of S100-positive cells as predictors of in vitro chondrogenicity of expanded human articular chondrocytes.J Cell Physiol.2010,222(2):411-420.
[35].Bottcher RT, Wiesner S, Braun A, et al. Profilin 1 is required for abscission during late cytokinesis of chondrocytes.EMBO J.2009,28(8):1157-1169.
[36].Hullerman E, Bijvelt JJ, Verkleij AJ, et al. Nuclear translocation of mitogen-activated protein kinase p42MAPK during the ongoing cell cycle.J Cell Physiol.1999,180(3):325-333.
[37].Ekholm SV, Zickert P, Reed SI, et al. Accumulation of cyclin E is not a prerequisite for passage through the restriction point.Mol Cell Biol.2001,21(9):3256-3265.
[38].Li J, Poi MJ, Tsai MD. Regulatory mechanisms of tumor suppressor P16INK4A and their relevance to cancer.Biochemistry.2011,50(25):5566-5582.
[39].Witkiewicz AK, Knudsen KE, Dicker AP, et al. The meaning of p16ink4a expression in tumors: functional significance, clinical associations and future developments.Cell Cycle.2011,10(15):2497-2503.
[40].许守明,陈双喜,何艳霞.细胞凋亡与细胞色素C[J].安徽农学通报.2009,15(15)：38-39.
[41].Grishko V, Xu M, Wilson G, et al. Apoptosis and mitochondrial dysfunction in human chondrocytes following exposure to lidocaine, bupivacaine, and ropivacaine.J Bone Joint Surg Am.2010,92(3):609-618.
[42]. Rosler S, Haase T, Claassen H, et al. Trefoil factor 3 is induced during degenerative and inflammatory joint disease, activates matrix metalloproteinases,and enhances apoptosis of articular cartilage chondrocytes.Arthritis Rheum.2010,62(3):815-825.
[43].Cory S, Adams JM. The Bcl-2 family:regulators of the cellular life-or-death switch. Nat Rev Cancer.2002,2(9):647-656.
[44].Saito M, Korsmeyer SJ, Schlesinger PH. BAX-dependent transport of cytochrome c reconstituted in pure liposomes.Nat Cell Biol.2000,2(8):553-555.
[45].Annis MG, Soucie EL, Dlugosz PJ, et al. Bax forms multispanning monomers that oligomerize to permeabilize membranes during apoptosis.EMBO J.2005,24(12):2096-2103.
[46].Kim PK, Annis MG, Dlugosz PJ, et al. During apoptosis Bcl-2 changes membrane topology at both the endoplasmic reticulum and mitochondria. Mol Cell.2004,14(4):523-529.
[47].Schendel SL, Montal M, Reed JC. Bcl-2 family proteins as ion-channels. Cell Death Differ.1998,5(5):372-380.
[48].Dlugosz PJ, Billen LP, Annis MG, et al. Bcl-2 changes conformation to inhibit Bax oligomerization.EMBO J.2006,25(11):2287-2296.
[49].Peng J, Tan C, Roberts GJ, et al. tBID elicits a conformational alteration in membrane-bound Bcl-2 such that it inhibits Bax pore formation.J Biol Chem.2006,281(47):35802-35811.
[50].Peng J, Ding J, Tan C, et al. Oligomerization of membrane-bound Bcl-2 is involved in its pore formation induced by tBid.Apoptosis.2009,14(10):1145-1153.
[51].黄继汪,黄晓晖,陈志扬,等.药理试验中动物间和动物与人体间的等效剂亮换算[J].中国临床药理学与治疗学.2004,9(9)：1069-1072.
[52].Soslow RA, Dannenberg AJ, Rush D, et al. Cox-2 is expressed in human pulmonary, colonic, and mammary tum ors.Cancer.2000,89(12):2637-2645.
[53].Havdrup T, Telhag H. Papain-induced changes in the knee joints of adult rabbits. Acta Orthop Scand.1977,48(2):143-149.
[54].Kopp S, Mejersjo C, Clemenssson E. Induction of osteoarthrosis in the guinea pig knee by papain.Oral Surg Oral Med Oral Pathol.1983,55(3):259-266.
[55].Marcelon G, Cros J, Guiraud R. Experimental model of osteoarthritis.Agents Actions.1976,6(1-3):191-194.
[56].Murat N, Karadam B, Ozkal S, et al. Quantification of papain-induced rat osteoarthritis in relation to time with the Mankin score.Acta Orthop Traumatol Turc.2007,41(3):233-237,2007.
[57].崔德娟.独活寄生汤临床应用心得[J].临床合理用药.2011,4(1)：67-68.
[58].顾程泷,朱艳伟,孙玉明.独活寄生汤在骨伤科的应用[J].河南中医,2013,33(4)：605-607.
[59].吴广文,褚剑锋,许惠风,等.独活寄生汤的药理作用及其在治疗骨性关节炎中的应用[J].中医正骨.2012,24(1)：37-39.
[60].Saied A, Cherin E, Gancher H, et al. Assessment of artieular-cartilage and subchondral bone.Bone Miner Res.1997,12(9):1378-1386.
[61].谈超,魏伟.骨关节炎的病因研究和治疗的进展[J].颈腰痛杂志.2002,23(2)：166-169.
[62].Giovannini S, Diaz-Romero J, Aigner T, et al. Population doublings and percentage of S100-positive cells as predictors of in vitro chondrogenicity of expanded human articular chondrocytes.J Cell Physiol.2010,222(2):411-420.
[63].Zhang M, Xie R, Hou W, et al. PTHrP prevents chondrocyte premature hypertrophy by inducing cyclin-Dl-dependent Runx2 and Runx3 phosphorylation,ubiquitylation and proteasomal degradation. J Cell Sci.2009,122(Pt 9):1382-1389.
[64].Hartweel LH, Weinert TA. Checkpoints:controls that ensure the order of cell cycle events. Science.1989,246(4930):629-634.
[65].薛丽香,李珺,张宗玉,等.细胞衰老与p16INK4a的转录调控[J].医学分子生物学杂.2006,3(1)：29-31.
[66].胡作为,周燕萍,沈自尹.p16基因与细胞衰老关系的研究进展[J].国外医学遗传学分册.2004,27(4)：200-202.
[67].Clurman BE, Roberts JM. Cell cycle and cancer. J Natl Cancer Inst.1995; 87:1499-1501.
[68].Hirama T, Koeffler HP. Role of the cyclin-dependent kinase inhibitors in the development of cancer.Blood.1995,86:841-854.
[69].Vivar O, Lin CL, Firestone GL, et al.3,3'-Diindolylmethane induces a G(1) arrest in human prostate cancer cells irrespective of androgen receptor and p53 Status. Biochem Pharmacol.2009,78(5):469-476.
[70].Herwig S, Strauss M. The retinoblastoma protein:a master regulator of cell cycle, differentiation and apoptosis.Eur J Biochem.1997,246(3):581-601.
[71].Grishko V, Xu M, Ho R, et al. Effects of hyaluronic acid on mitochondrial function and mitochondria-driven apoptosis following oxidative stress in human chondrocytes. J Biol Chem.2009,284(14):9132-9139.
[72].Milner PI, Wilkins RJ, Gibson JS. The role of mitochondrial reactive oxygen species in pH regulation in articular chondrocytes.Osteoarthritis Cartilage.2007,15(7):735-742.
[73].Teixeira CC, Costas AP, Nemelivsky Y. Apoptosis of growth plate chondrocytes occurs through a mitochondrial pathway.Angle Orthod.2007,77(1):129-134.
[74].Ruiz-Romero C, Lopez-Armada MJ, Blanco FJ. Mitochondrial proteomic charactetization of human normal articular chondrocytes.Osteoarthritis Cartilage.2006,14(6):507-518.
[75].Pabbruwe MB, Stewart K, Chaudhuri JB. A comparison of colorimetrix and DNA quantification assays for the assessment of meniscal fibrochondrocyte proliferation in microcarrier culture.Biotechnol Lett.2005,27(19):1451-1455.
[76].Blanco FJ, Lopez-Armada MJ, Maneiro E. Mitochondrial dysfunction in osteoarthritis. Mitochondrion.2004,4(5-6):715-728.
[77].Maneiro E, Lopez-Armada MJ, de Andres MC, et al. Effect of nitric oxide on mitochondrial respiratory activity of human articular chondrocytes. Ann Rheum Dis.2005,64(3):388-395.
[78].Ma Q, Li X, Vale-Cruz D, et al. Activating transcription factor 2 controls Bcl-2 promoter activity in growth plate chondrocytes.J Cell Biochem.2007,101(2):477-487.
[79].Oshima Y, Akiyama T, Hikita A, et al. Pivotal role of Bcl-2 family proteins in the regulation of chondrocyte apoptosis. J Biol Chem.2008,283(39):26499-26508.
[80].Surendran S, Kim SH, Jee BK, et al. Anti-apoptotic Bcl-2 gene transfection of human articular chondrocytes protects against nitric oxide-induced apoptosis. J Bone Joint Surg Br.2006,88(12):1660-1665.
[81].Pucci B, Adams CS, Fertala J, et al. Development of the terminally differentiated state sensitizes epiphyseal chondrocytes to apoptosis through caspase-3 activation. J Cell Physiol.2007,210(3):609-615.
[82].Shakibaei M, John T, Seifarth C, et al. Resveratrol inhibits IL-1 beta-induced stimulation of caspase-3 and cleavage of PARP in human articular chondrocytes in vitro. Ann N Y Sci.2007,1095:554-563.
[83].彭程,肖涛,罗远明,等.IGF-1对IL-1诱导的兔关节软骨细胞NO和PGE2的影响[J].中南大学学报(医学版).2008,33(3)：197-203.
[84].Park K, Hoffmeister B, Han DK and Hasty K. Therapeutic ultrasound effects on interleukin-1β stimulated cartilage construct in vitro. Ultrasound Med Biol.2007,33(2):286-2957.
[85].Mantymaa P, Siitonen T, Guttorm T, et al. Induction of mitochondrial manganese superoxide dismutase confers resistance to apoptosis in acute myeloblastic leukaemia cells exposed to etoposide. Br J Haematol.2000,108(3):574-581.
[86].Korper S, Nolte F, Rojewski MT, et al. The K+ channel openers diazoxide and NS1619 induce depolarization of mitochondria and have differential effects on cell Ca2+ in CD34+cell line KG-la.Exp Hematol.2003,31 (9):815-823.
[87].Chockalingam PS, Varadarajan U, Sheldon R, et al. Involvement of protein kinase Czeta in interleukin-1 beta induction of ADAMTS-4 and type 2 nitric oxide synthase via NF-kappa B signaling in primary human osteoarthritis chondrocytes.Arthritis Rheum.2007,56(12):4074-4083.
[88].Ho LJ, Lin LC, Hung LF, et al. Retinoic acid blocks pro-inflammatory cytokine-induced matrix metalloproteinase production by down-regulating JNK-AP-1 signaling in human chondrocytes.Biochem Pharmacol.2005,70(2):200-208.
[89].陈文彬,王有赤.诊断学[M].5版.北京：人民卫生出版社,2001：380-381.
[90].张艳,柴岗,刘伟,等.体外培养过程中去分化的人软骨细胞基因表达谱的变化[J].中华整形外科杂志.2007,23(4)：331-334.
[91].Ryu JH, Chun JS. Opposing roles of WNT-5A and WNT-11 in interleukin-1 beta regulation of type II collagen expression in articular chondrocytes.J Bilo Chem.2006,281(31):22039-22047.
[92].张莉,郭全义,眭翔,等.羊软骨细胞在生物反应器中的培养和扩增[J].中国矫形外科杂志.2006,14(6)：446-448.
[93].Gosset M, Berenbaum F, Thirion S, et al. Primary culture and phenotyping of murine chondrocytes.Nat Protoc.2008,3(8):1253-1260.
[94].Irie Y, Mizumoto H, Fujino S, et al. Reconstruction of cartilage tissue using scaffold-free organoid culture technique.J Biosci Bioeng.2008,105(5):450-453.
[95].刘小荣,张笠,高武,等.新西兰兔关节软骨细胞分离培养与鉴定的实验研究[J].国际检验医学杂志.2012,33(19)：2307-2312.
[96].Karsdal MA, Sumer EU, Wulf H, et al. Induction of increased cAMP levels in articular chondrocytes blocks matrix metalloproteinase-mediated cartilage degradation, but not aggrecanase-mediated cartilage degradation.Arthritis Rheum.2007,56(5):1549-1558.
[97].Seki K, Fujimori T, Savagner P, et al. Mouse Snail family transcription repressors regulate chondrocyte, extracellular matrix, type II collagen, and aggrecan.Am J Pathol.2003,278(43):41862-41870.
[98].Enomoto H, Inoki I, Komiya K, et al. Vascular endothelial growth factor isoforms and their receptors are expressed in human osteoarthritis.Am J Pathol.2003,162(1):171-181.
[99].Khan AA, Suits JM, Kandel RA, et al. The effect of continuous culture on the growth and structure of tissue-engineered cartilage.Biotechnol Prog.2009,25(2):508-515.
[100].Kaushik AP, Martin JA, Zhang Q, et al. Cartilage abnormalities associated with defects of chondrocytic primary clilia in Bardet-Biedl syndrome mutant mice.J Orthop Res.2009,27(8):1093-1099.
[101].刘振峰,方锐,孟庆才.Ⅱ型胶原酶消化法可短时间获得大量纯化大鼠关节软骨细胞[J].中国组织工程研究与临床康复.2011,15(50)：9323-9326.
[102].Iwama H, Amagaya S, Ogihara Y. Effect of shosaikoto, a Japanese and Chinese traditional herbal medical mixture, on the mitogenic activity of lipopolysaccharide:a new pharmacological testing method.J Ethnopharmacol.1987,21(1):45-53.
[104].张群豪,陈可冀.血清药理学在中药及复方研究中应用的评价[J].中国中西医结合杂志.1996,16(3)：131.
[105].王筠,张军平.中药血清药理学实验方法研究概况[J].天津中医药.2005,22(1)：86-88.
[106].杜新亮,隋峰,张畅斌,等.桑菊饮含药血清对小鼠巨噬细胞Toll样受体表达的影响[J].中国实验方剂学杂志.2010,16(1)：57-61.
[107].韩林.中药血清药理学研究方法探讨与思考[J].浙江中西医结合杂志.2009,19(5)：319-322.
[108].崔晓兰,贺玉琢,高英杰,等.中药复方血清药理研究方法学探讨-I[J].中国实验方剂学杂志.1998,4(2)：13-15.
[109].赵保胜,李兰芳,马悦颖,等.参苏饮含药血清对小鼠巨噬细胞Toll样受体及其下游信号转导通路的影响[J].中国中药杂志.2007,32(4)：327-332.
[110].Lyons-Giordano B, Pratta MA, et al. Interleukin-1 differentially modulates chondrocyte expression of cycloxygenase-2 and phospholipase A2.Exp Cell Res.1993,206(1):58-62.
[111].焦岩涛,王大章,胡静.白介素-1及地塞米松对人髁状突软骨细胞增殖、代谢及前胶原基因表达的影响[J].中华创伤杂志.2000,16(2)：94-95.
[112].潘海乐,王耶.IL-1与骨性关节炎研究新进展[J].中国老年性杂志.2002,22(1)：76-78.
[113].刘世清,陈庆,杜予民,等.羧甲基壳聚糖对白细胞介素-1β刺激的软骨细胞增殖能力和表型的保护作用[J].中华风湿病学杂志.2005,9(11)：672-676.
[114].苏友新,林学义,陈秀明,等.IL-1β对大鼠关节软骨细胞"Wnt/0-catenin"通路的影响[J].福建中医药.2012,43(6)：46-48.
[115].Christelle S, Maranne MH, Michelle AD, et al. Effects of rhein on human articular chondrocytes in alginate beads. Bioche Pharmacology.2003,65(3):377-388.
[116].Wu SQ, Otero M, Unger FM, et al. Anti-inflammatory activity of an ethanolic Caesalpinia sappan extract in human chondrocytes and macrophagesJournal of Ethnopharmacology.2011,138(2):364-372.
[117].Sanchez C, Mathy-Hartert M, Deberg MA, et al. Effects of rhein on human articular chondrocytes in alginate beads. Biochem Pharmacol.2003,65(3):377-388.
[118].Kusuzaki K, Sugimoto S, Takeshita H, et al. DNA cytofluorometric analysis of chondrocytes in human articular cartilages under normal aging or arthritic conditions. Osteoarthritis Cartilage.2001,9(7):664-670.
[119].Duman-Scheel M, Weng L, Xin S, et al. Hedgehog regulates cell growth and proliferation by inducing cyclin D and cyclin E. Nature.2002,417(6886):299-304.
[120].Cossarizza A, Baccarani-Contri M, Kalashnikova G, et al. A new method for the cytofluorimetric analysis of mitochondrial membrane potential using the J-aggregate forming lipophilic cation 5,5',6,6'-tetrachloro-1,1',3,3' tetraeth-ylbenzimidazolylcarbocyanine iodide (JC-1).Biochem Biophys Res Commun.1993,197(1):40-45.
[121].Smiley ST, Reers M, Mottola-Hartshom C, et al. Intracellular heterogeneity in mitochondrial membrane potentials revealed by a J-aggregate forming lipophilic cation JC-1. Proc Natl Acad Sci USA.1991,88(9):3671-3675.
[1].王爱武,刘娅,雒琪,等.独活寄生汤抗炎、镇痛作用的药效学研究[J].中国实验方剂学杂志,2008,14(12)：61-64.
[2].赵玉珍,傅正宗,卢春凤,等.复方独活寄生合剂抗炎、镇痛作用观察[J].黑龙江医药科学,2003,26(3)：3-4.
[3].方剑乔,刘金洪,赵天征,等.独活寄生汤对小鼠胶原性关节炎治疗作用的研究[J].中国中医药科技,2000,7(5)：289-290.
[4].刘英杰,马利杰,王相利.独活寄生汤对兔膝骨性关节炎关节液白细胞介素-1和肿瘤坏死因子的影响[J].河北中医,2007,29(8)：748-750.
[5],李爱萍,何昌谋.独活寄生汤对兔膝骨关节炎体液中NO、SOD水平的影响[J].陕西中医,2010,31(10)：1430-1431.
[6].程维,张玉辉.独活寄生汤对膝骨性关节炎患者关节滑液中细胞因子的影响[J].辽宁中医药大学学报,2011,13(10)：216-217.
[7].朱自平.独活寄生汤对微循环的影响[J].中成药,1991,13(3)：26.
[8].许青嫒,谢人明,任军鹏等.独活寄生汤对麻醉动物脑循环的作用[J].陕西中医,1989,10(9)：425.
[9].陈筑红,李悦,杨凡,等.独活寄生汤对激素性股骨头缺血坏死的治疗作用[J].现代中西医结合杂志,2006,15(11)：1493.
[10].张若楠,王三虎,任东青.独活寄生汤对荷瘤小鼠的抗肿瘤作用研究[J].中国实验方剂学杂志,2007,13(10)：28-31.
[11].段泾云.独活寄生汤抗炎免疫药理作用研究[J].中成药研究,1988,(5)：28.
[12].王爱武,刘娅,林晓燕,等.独活寄生汤及其配方颗粒对胶原诱导型关节炎大鼠滑膜组织病理学变化的影响[J].中国中医药科技,2010,17(3)：202-203.
[13]].孙学东,姚华,姚杰.独活寄生汤加减治疗膝骨关节炎[J].北京中医药,2008,27(5)：362-363.
[14].李宝春.独活寄生汤加减治疗膝关节骨性关节炎100例临床观察[J].中医中药,2010,17(15)：80-81.
[15]].周友连.独活寄生汤加减治疗膝关节骨性关节炎116例疗效观察[J].浙江中医杂志,2007,42(10)：591.
[16].余建华,张衡.独活寄生汤治疗膝骨关节炎临床观察[J].中国实验方剂学杂志,2010,16(7)：215-217.
[17].高云.独活寄生汤加减治疗膝关节骨性关节炎临床疗效分析[J].中医中药,2010,7(20)：73.
[18].岑祖怡,周剑鹏.独活寄生汤治疗膝关节骨性关节炎50例疗效观察[J].新中医,2010,42(8)：50-51.
[19]].顾玉潜.独活寄生汤治疗膝关节骨性关节炎60例疗效观察[J].长春中医药大学学报,2008,24(2)：202-203.
[20].朱健儿.独活寄生汤治疗退行性膝骨关节炎60例[J].浙江中医学院学报,2005,29(4)：40.
[21].李培豪.独活寄生汤配合玻璃酸钠关节腔注射治疗膝骨性关节炎86例[J].中国中医急症,2010,19(7)：1236.
[22].管亦坚.关节腔内注射配合独活寄生汤治疗膝骨关节炎滑膜积液疗效观察[J].浙江中医药大学学报,2010,34(3)：399.
[23].于泽来,王伟,林一峰.独活寄生汤配合局部手法理伤治疗膝关节退行性骨关节炎[J].中国临床康复,2005,9(38)：124-125.
[24].邓泽前,张荣龙,俞宙.中药配合理疗手法治疗膝关节骨性关节炎106例分析[J].中国误诊学杂志,2008,8(34)：8466-8467.
[25].江飞,张清文,贺飞,等.独活寄生汤配合针刺治疗膝关节骨性炎症[J].中华全科医学,2010,8(9)：1156-1157.
[26].杨梦林,华刚,刘守新.中药加针灸治疗膝关节骨性关节炎164例[J].吉林中医药,2007,27(2)：41.
[27].汪利合.独活寄生汤加减结合运动疗法治疗膝骨关节炎的临床研究[J].中医正骨,2011,23(9)：3-5.
[28].刘俊峰,张卫华.独活寄生汤加减配合中药薰蒸治膝骨性关节炎50例[J].江西中医药,2011,42(4)：42-44.
[29].王武炼,叶锦霞,刘献祥.独活寄生汤加减内服外洗治疗膝骨性关节炎66例临床观察[J].福建中医药大学学报,2011,21(2)：44-45.
[30].李振荣,吕长青.独活寄生汤加减内服外用治疗膝骨关节炎36例[J].陕西中医,2011,32(9)：1168-1169.
[31].徐新玉,包力,韩明霞,等.独活寄生汤合中药离子导入疗膝关节骨性关节炎83例[J].中国中医药科技,2005,12(4)：259.
[32].杨芳,王月琴.独活寄生汤加减配合中药雾化治疗骨性关节炎163例[J].现代中医药,2007,27(5)：23-24.
[33].钟利进,陈懋生.独活寄生汤联合维生素D3治疗膝关节骨性关节炎的疗效[J].中医药导报,2009,15(9)：22-23.
[34].刘志文,赵淑平,陈茵,等.天灸联合独活寄生汤治疗膝骨关节炎96例[J].广东医学,2010,31(13)：1754-1755.
[2].李西海,梁文娜.软骨下骨骨重塑与骨关节炎的关系[J].国际骨科学杂志.2007,28(1)：35-37.
[3].Dahm DL, Al-Rayashi W, Dajani K, et al. Patellofemoral arthroplasty versus total knee arthroplasty in patients with isolated patellofemoral osteoarthritis.Am J Orthop (Belle Mead NJ).2010,39(10):487-491.
[4].Vuolteenaho K, Moilanen T, Knowles RG, et al. The role of nitric oxide in osteoarthritis. Scand J Rheumatol.2007,36(4):247-258.
[5].Shakoor MA, Rahman MS, Azad AK, et al. Effects of isometric quadriceps muscle strengthening exercise on chronic osteoarthritis of the knee.Bangladesh Med Res Counc Bull.2010,36(1):20-22.
[6].李宁华,薛庆云,张毅,等.中国六城市中老年人群X线膝骨关节炎流行病学分析[J].实用医学杂志.2008,24(16)：2887-2888.
[7].周青,王宇仁,周之德,等.不同分子量的透明质酸钠治疗膝骨关节炎的比较[J].上海第二医科大学学报.2000,20(4)：361-363.
[8].邱贵兴.骨关节炎流行病学和病因学进展[J].继续医学教育.2005,19(7)：68-69.
[9].周肃陵,刘肖瑜,王正.独活寄生汤在骨伤科的临床应用[J].中医正骨.2006,18(12)：67.
[10].李西海,陈文列,刘献祥.补肾柔肝法防治骨性关节炎作用机制探讨[J].福建中医药大学学报.2011,21(2)：66-69.
[11].郑春松,叶蕻芝,许惠凤,等.分子对接法预测独活寄生汤对INF-α和IL-1β作用的活性成分[J].福建中医药大学学报.2012,22(4)：28-30.
[12].王武炼,叶锦霞,刘献祥,等.独活寄生汤加减内服外洗治疗膝骨性关节炎66例临床观察[J].福建中医药大学学报.2011,21(2)：44-45.
[13].李西海,张翼,叶蕻芝,等.独活寄生汤治疗凤寒湿痹型膝骨性关节炎的机制探讨[J].中医正骨.2012,24(1)：68-71.
[14].吴广文,褚剑锋,许惠风,等.独活寄生汤的药理作用及其在治疗骨性关节炎中的应用[J].中医正骨.2012,24(1)：37-39.
[15].郑春松,叶蕻芝,许惠凤,等.从化学空间角度看独活寄生汤治疗骨性关节炎的多靶点作用[J].福建中医药大学学报.2011,21(5)：35-37.
[16].Zheng CS, X XJ, Y HZ, et al. Computational approaches for exploring the potential synergy and polypharmacology of Duhuo Jisheng Decoction in the therapy of osteoarthritis. Mol Med Report.2013,7(6):1812-1818.
[17].Haq I, Murphy E, Dacre J. Osteoarthritis. Postgrad Med J.2003,79(933):377-383.
[18].Uryu N, Okada K, Kawakita K. Analgesic effects of indirect moxibustion on an experimental rat model od osteoarthritis in the knee.Acupunct Med.2007,25(4):175-183.
[19].Torelli SR, Rahal SC, Volpi RS, et al. Histopathological evaluation of treatment with chondroitin sulphate for osteoarthritis induced by continuous immobilization in rabbits. J Vet Med A Physiol Pathol Clin Med.2005,52(1):45-51.
[20].Legendre F, Heuze A, Boukerrouche K, et al. Rhein, the metabolite of diacerhein, reduces the proliferation of osteoarthritic chondrocytes and synoviocytes without inducing apoptosis.Scand J Rheumatol.2009,38(2):104-111.
[21].Matsumoto T, Cooper GM, Gharaibeh B, et al. Cartilage repair in a rat model of osteoarthritis through intraarticular transplantation of muscle-derived stem cells expressing bone morphogenetic protein 4 and soluble flt-1.Arthritis Rheum.2009,30(5):1390-1405.
[22].Guo D, Ding L, Homandberg GA. Telopeptides of type Ⅱ collagen upregulate proteinases and damage cartilage but are less effective than highly active fibronectin fragments. Inflamm Res.2009,58(3):161-169.
[23].Li X, Ellman M, Muddasani P, et al. Prostaglandin E2 and its cognate EP receptors control human adult articular cartilage homeostasis and are linked to the pathophysiology of osteoarthritis.Arthritis Rheum.2009,30(2):513-523.
[24].Yang X, Chen L, Xu X, et al. TGF-β/Smad3 signals repress chondrocyte hypertrophic differentiation and are required for maintaining articular cartilage.J Cell Biol.2001,153(1):35-46.
[25].Iannone F, De Bari C, Scioscia C, et al. Increased Bcl-2/p53 ratio in human osteoarthritic cartilage:a possible role in regulation of chondrocyte metabolism.Ann Rheum Dis.2005,64(2):217-221.
[26].Zhou HW, Lou SQ, Zhang K. Recovery of function in osteoarthritic chondrocytes induced by p16INK4a-siRNA in vitr. Rheumatology(Oxford).2004,43(5):555-568.
[27].Tetlow LC, Woolley DE. Histamine stimulates the proliferation of human articular chondrocytes in vitro and is expressed by chondrocytes in osteoarthritic cartilage.Ann Rheum Dis.2003,62(10):991-994.
[28].Sharma L, Kapoor D, Issa S. Epidemiology of osteoarthritis:an update.Curr Opin Rheumatol.2006,18(2):147-156.
[29].Sandell LJ, Aigner T. Articular cartilage and changes in arthritis. An introduction:Cell biology of osteoarthritis.Arthritis Res.2001,3(2):107-113.
[30].Malemud CJ. Protein kinases in chondrocyte signaling and osteoarthritis. Clin Orthop Relat Res.2004,(427 Supp 1):145-151.
[31].Mistry D, Oue Y, Chambers MG, et al. Chondrocyte death during murine osteoarthritis[J]. Osteoarthritis Cartilage.2004,12(2):131-141.
[32].Embree MC, Kilts TM, Ono M, et al. Biglycan and fibromodulin have essential roles in regulating chondrogenesis and extracellular matrix turnover in temporomandibular joint osteoarthritis.Am J Pathol.2010,176(2):812-826.
[33].Kunitomo T, Takahashi KA, Arai Y, et al. Influence of extracellular matrix on the expression of inflammatory cytokines, proteases, and apoptosis-related genes induced by hydrostatic pressure in three-dimensionally cultured chondrocytes.J Orthop Sci.2009,14(6):776-783.
[34].Giovannini S, Diaz-Romero J, Aigner T, et al. Population doublings and percentage of S100-positive cells as predictors of in vitro chondrogenicity of expanded human articular chondrocytes.J Cell Physiol.2010,222(2):411-420.
[35].Bottcher RT, Wiesner S, Braun A, et al. Profilin 1 is required for abscission during late cytokinesis of chondrocytes.EMBO J.2009,28(8):1157-1169.
[36].Hullerman E, Bijvelt JJ, Verkleij AJ, et al. Nuclear translocation of mitogen-activated protein kinase p42MAPK during the ongoing cell cycle.J Cell Physiol.1999,180(3):325-333.
[37].Ekholm SV, Zickert P, Reed SI, et al. Accumulation of cyclin E is not a prerequisite for passage through the restriction point.Mol Cell Biol.2001,21(9):3256-3265.
[38].Li J, Poi MJ, Tsai MD. Regulatory mechanisms of tumor suppressor P16INK4A and their relevance to cancer.Biochemistry.2011,50(25):5566-5582.
[39].Witkiewicz AK, Knudsen KE, Dicker AP, et al. The meaning of p16ink4a expression in tumors: functional significance, clinical associations and future developments.Cell Cycle.2011,10(15):2497-2503.
[40].许守明,陈双喜,何艳霞.细胞凋亡与细胞色素C[J].安徽农学通报.2009,15(15)：38-39.
[41].Grishko V, Xu M, Wilson G, et al. Apoptosis and mitochondrial dysfunction in human chondrocytes following exposure to lidocaine, bupivacaine, and ropivacaine.J Bone Joint Surg Am.2010,92(3):609-618.
[42]. Rosler S, Haase T, Claassen H, et al. Trefoil factor 3 is induced during degenerative and inflammatory joint disease, activates matrix metalloproteinases,and enhances apoptosis of articular cartilage chondrocytes.Arthritis Rheum.2010,62(3):815-825.
[43].Cory S, Adams JM. The Bcl-2 family:regulators of the cellular life-or-death switch. Nat Rev Cancer.2002,2(9):647-656.
[44].Saito M, Korsmeyer SJ, Schlesinger PH. BAX-dependent transport of cytochrome c reconstituted in pure liposomes.Nat Cell Biol.2000,2(8):553-555.
[45].Annis MG, Soucie EL, Dlugosz PJ, et al. Bax forms multispanning monomers that oligomerize to permeabilize membranes during apoptosis.EMBO J.2005,24(12):2096-2103.
[46].Kim PK, Annis MG, Dlugosz PJ, et al. During apoptosis Bcl-2 changes membrane topology at both the endoplasmic reticulum and mitochondria. Mol Cell.2004,14(4):523-529.
[47].Schendel SL, Montal M, Reed JC. Bcl-2 family proteins as ion-channels. Cell Death Differ.1998,5(5):372-380.
[48].Dlugosz PJ, Billen LP, Annis MG, et al. Bcl-2 changes conformation to inhibit Bax oligomerization.EMBO J.2006,25(11):2287-2296.
[49].Peng J, Tan C, Roberts GJ, et al. tBID elicits a conformational alteration in membrane-bound Bcl-2 such that it inhibits Bax pore formation.J Biol Chem.2006,281(47):35802-35811.
[50].Peng J, Ding J, Tan C, et al. Oligomerization of membrane-bound Bcl-2 is involved in its pore formation induced by tBid.Apoptosis.2009,14(10):1145-1153.
[51].黄继汪,黄晓晖,陈志扬,等.药理试验中动物间和动物与人体间的等效剂亮换算[J].中国临床药理学与治疗学.2004,9(9)：1069-1072.
[52].Soslow RA, Dannenberg AJ, Rush D, et al. Cox-2 is expressed in human pulmonary, colonic, and mammary tum ors.Cancer.2000,89(12):2637-2645.
[53].Havdrup T, Telhag H. Papain-induced changes in the knee joints of adult rabbits. Acta Orthop Scand.1977,48(2):143-149.
[54].Kopp S, Mejersjo C, Clemenssson E. Induction of osteoarthrosis in the guinea pig knee by papain.Oral Surg Oral Med Oral Pathol.1983,55(3):259-266.
[55].Marcelon G, Cros J, Guiraud R. Experimental model of osteoarthritis.Agents Actions.1976,6(1-3):191-194.
[56].Murat N, Karadam B, Ozkal S, et al. Quantification of papain-induced rat osteoarthritis in relation to time with the Mankin score.Acta Orthop Traumatol Turc.2007,41(3):233-237,2007.
[57].崔德娟.独活寄生汤临床应用心得[J].临床合理用药.2011,4(1)：67-68.
[58].顾程泷,朱艳伟,孙玉明.独活寄生汤在骨伤科的应用[J].河南中医,2013,33(4)：605-607.
[59].吴广文,褚剑锋,许惠风,等.独活寄生汤的药理作用及其在治疗骨性关节炎中的应用[J].中医正骨.2012,24(1)：37-39.
[60].Saied A, Cherin E, Gancher H, et al. Assessment of artieular-cartilage and subchondral bone.Bone Miner Res.1997,12(9):1378-1386.
[61].谈超,魏伟.骨关节炎的病因研究和治疗的进展[J].颈腰痛杂志.2002,23(2)：166-169.
[62].Giovannini S, Diaz-Romero J, Aigner T, et al. Population doublings and percentage of S100-positive cells as predictors of in vitro chondrogenicity of expanded human articular chondrocytes.J Cell Physiol.2010,222(2):411-420.
[63].Zhang M, Xie R, Hou W, et al. PTHrP prevents chondrocyte premature hypertrophy by inducing cyclin-Dl-dependent Runx2 and Runx3 phosphorylation,ubiquitylation and proteasomal degradation. J Cell Sci.2009,122(Pt 9):1382-1389.
[64].Hartweel LH, Weinert TA. Checkpoints:controls that ensure the order of cell cycle events. Science.1989,246(4930):629-634.
[65].薛丽香,李珺,张宗玉,等.细胞衰老与p16INK4a的转录调控[J].医学分子生物学杂.2006,3(1)：29-31.
[66].胡作为,周燕萍,沈自尹.p16基因与细胞衰老关系的研究进展[J].国外医学遗传学分册.2004,27(4)：200-202.
[67].Clurman BE, Roberts JM. Cell cycle and cancer. J Natl Cancer Inst.1995; 87:1499-1501.
[68].Hirama T, Koeffler HP. Role of the cyclin-dependent kinase inhibitors in the development of cancer.Blood.1995,86:841-854.
[69].Vivar O, Lin CL, Firestone GL, et al.3,3'-Diindolylmethane induces a G(1) arrest in human prostate cancer cells irrespective of androgen receptor and p53 Status. Biochem Pharmacol.2009,78(5):469-476.
[70].Herwig S, Strauss M. The retinoblastoma protein:a master regulator of cell cycle, differentiation and apoptosis.Eur J Biochem.1997,246(3):581-601.
[71].Grishko V, Xu M, Ho R, et al. Effects of hyaluronic acid on mitochondrial function and mitochondria-driven apoptosis following oxidative stress in human chondrocytes. J Biol Chem.2009,284(14):9132-9139.
[72].Milner PI, Wilkins RJ, Gibson JS. The role of mitochondrial reactive oxygen species in pH regulation in articular chondrocytes.Osteoarthritis Cartilage.2007,15(7):735-742.
[73].Teixeira CC, Costas AP, Nemelivsky Y. Apoptosis of growth plate chondrocytes occurs through a mitochondrial pathway.Angle Orthod.2007,77(1):129-134.
[74].Ruiz-Romero C, Lopez-Armada MJ, Blanco FJ. Mitochondrial proteomic charactetization of human normal articular chondrocytes.Osteoarthritis Cartilage.2006,14(6):507-518.
[75].Pabbruwe MB, Stewart K, Chaudhuri JB. A comparison of colorimetrix and DNA quantification assays for the assessment of meniscal fibrochondrocyte proliferation in microcarrier culture.Biotechnol Lett.2005,27(19):1451-1455.
[76].Blanco FJ, Lopez-Armada MJ, Maneiro E. Mitochondrial dysfunction in osteoarthritis. Mitochondrion.2004,4(5-6):715-728.
[77].Maneiro E, Lopez-Armada MJ, de Andres MC, et al. Effect of nitric oxide on mitochondrial respiratory activity of human articular chondrocytes. Ann Rheum Dis.2005,64(3):388-395.
[78].Ma Q, Li X, Vale-Cruz D, et al. Activating transcription factor 2 controls Bcl-2 promoter activity in growth plate chondrocytes.J Cell Biochem.2007,101(2):477-487.
[79].Oshima Y, Akiyama T, Hikita A, et al. Pivotal role of Bcl-2 family proteins in the regulation of chondrocyte apoptosis. J Biol Chem.2008,283(39):26499-26508.
[80].Surendran S, Kim SH, Jee BK, et al. Anti-apoptotic Bcl-2 gene transfection of human articular chondrocytes protects against nitric oxide-induced apoptosis. J Bone Joint Surg Br.2006,88(12):1660-1665.
[81].Pucci B, Adams CS, Fertala J, et al. Development of the terminally differentiated state sensitizes epiphyseal chondrocytes to apoptosis through caspase-3 activation. J Cell Physiol.2007,210(3):609-615.
[82].Shakibaei M, John T, Seifarth C, et al. Resveratrol inhibits IL-1 beta-induced stimulation of caspase-3 and cleavage of PARP in human articular chondrocytes in vitro. Ann N Y Sci.2007,1095:554-563.
[83].彭程,肖涛,罗远明,等.IGF-1对IL-1诱导的兔关节软骨细胞NO和PGE2的影响[J].中南大学学报(医学版).2008,33(3)：197-203.
[84].Park K, Hoffmeister B, Han DK and Hasty K. Therapeutic ultrasound effects on interleukin-1β stimulated cartilage construct in vitro. Ultrasound Med Biol.2007,33(2):286-2957.
[85].Mantymaa P, Siitonen T, Guttorm T, et al. Induction of mitochondrial manganese superoxide dismutase confers resistance to apoptosis in acute myeloblastic leukaemia cells exposed to etoposide. Br J Haematol.2000,108(3):574-581.
[86].Korper S, Nolte F, Rojewski MT, et al. The K+ channel openers diazoxide and NS1619 induce depolarization of mitochondria and have differential effects on cell Ca2+ in CD34+cell line KG-la.Exp Hematol.2003,31 (9):815-823.
[87].Chockalingam PS, Varadarajan U, Sheldon R, et al. Involvement of protein kinase Czeta in interleukin-1 beta induction of ADAMTS-4 and type 2 nitric oxide synthase via NF-kappa B signaling in primary human osteoarthritis chondrocytes.Arthritis Rheum.2007,56(12):4074-4083.
[88].Ho LJ, Lin LC, Hung LF, et al. Retinoic acid blocks pro-inflammatory cytokine-induced matrix metalloproteinase production by down-regulating JNK-AP-1 signaling in human chondrocytes.Biochem Pharmacol.2005,70(2):200-208.
[89].陈文彬,王有赤.诊断学[M].5版.北京：人民卫生出版社,2001：380-381.
[90].张艳,柴岗,刘伟,等.体外培养过程中去分化的人软骨细胞基因表达谱的变化[J].中华整形外科杂志.2007,23(4)：331-334.
[91].Ryu JH, Chun JS. Opposing roles of WNT-5A and WNT-11 in interleukin-1 beta regulation of type II collagen expression in articular chondrocytes.J Bilo Chem.2006,281(31):22039-22047.
[92].张莉,郭全义,眭翔,等.羊软骨细胞在生物反应器中的培养和扩增[J].中国矫形外科杂志.2006,14(6)：446-448.
[93].Gosset M, Berenbaum F, Thirion S, et al. Primary culture and phenotyping of murine chondrocytes.Nat Protoc.2008,3(8):1253-1260.
[94].Irie Y, Mizumoto H, Fujino S, et al. Reconstruction of cartilage tissue using scaffold-free organoid culture technique.J Biosci Bioeng.2008,105(5):450-453.
[95].刘小荣,张笠,高武,等.新西兰兔关节软骨细胞分离培养与鉴定的实验研究[J].国际检验医学杂志.2012,33(19)：2307-2312.
[96].Karsdal MA, Sumer EU, Wulf H, et al. Induction of increased cAMP levels in articular chondrocytes blocks matrix metalloproteinase-mediated cartilage degradation, but not aggrecanase-mediated cartilage degradation.Arthritis Rheum.2007,56(5):1549-1558.
[97].Seki K, Fujimori T, Savagner P, et al. Mouse Snail family transcription repressors regulate chondrocyte, extracellular matrix, type II collagen, and aggrecan.Am J Pathol.2003,278(43):41862-41870.
[98].Enomoto H, Inoki I, Komiya K, et al. Vascular endothelial growth factor isoforms and their receptors are expressed in human osteoarthritis.Am J Pathol.2003,162(1):171-181.
[99].Khan AA, Suits JM, Kandel RA, et al. The effect of continuous culture on the growth and structure of tissue-engineered cartilage.Biotechnol Prog.2009,25(2):508-515.
[100].Kaushik AP, Martin JA, Zhang Q, et al. Cartilage abnormalities associated with defects of chondrocytic primary clilia in Bardet-Biedl syndrome mutant mice.J Orthop Res.2009,27(8):1093-1099.
[101].刘振峰,方锐,孟庆才.Ⅱ型胶原酶消化法可短时间获得大量纯化大鼠关节软骨细胞[J].中国组织工程研究与临床康复.2011,15(50)：9323-9326.
[102].Iwama H, Amagaya S, Ogihara Y. Effect of shosaikoto, a Japanese and Chinese traditional herbal medical mixture, on the mitogenic activity of lipopolysaccharide:a new pharmacological testing method.J Ethnopharmacol.1987,21(1):45-53.
[104].张群豪,陈可冀.血清药理学在中药及复方研究中应用的评价[J].中国中西医结合杂志.1996,16(3)：131.
[105].王筠,张军平.中药血清药理学实验方法研究概况[J].天津中医药.2005,22(1)：86-88.
[106].杜新亮,隋峰,张畅斌,等.桑菊饮含药血清对小鼠巨噬细胞Toll样受体表达的影响[J].中国实验方剂学杂志.2010,16(1)：57-61.
[107].韩林.中药血清药理学研究方法探讨与思考[J].浙江中西医结合杂志.2009,19(5)：319-322.
[108].崔晓兰,贺玉琢,高英杰,等.中药复方血清药理研究方法学探讨-I[J].中国实验方剂学杂志.1998,4(2)：13-15.
[109].赵保胜,李兰芳,马悦颖,等.参苏饮含药血清对小鼠巨噬细胞Toll样受体及其下游信号转导通路的影响[J].中国中药杂志.2007,32(4)：327-332.
[110].Lyons-Giordano B, Pratta MA, et al. Interleukin-1 differentially modulates chondrocyte expression of cycloxygenase-2 and phospholipase A2.Exp Cell Res.1993,206(1):58-62.
[111].焦岩涛,王大章,胡静.白介素-1及地塞米松对人髁状突软骨细胞增殖、代谢及前胶原基因表达的影响[J].中华创伤杂志.2000,16(2)：94-95.
[112].潘海乐,王耶.IL-1与骨性关节炎研究新进展[J].中国老年性杂志.2002,22(1)：76-78.
[113].刘世清,陈庆,杜予民,等.羧甲基壳聚糖对白细胞介素-1β刺激的软骨细胞增殖能力和表型的保护作用[J].中华风湿病学杂志.2005,9(11)：672-676.
[114].苏友新,林学义,陈秀明,等.IL-1β对大鼠关节软骨细胞"Wnt/0-catenin"通路的影响[J].福建中医药.2012,43(6)：46-48.
[115].Christelle S, Maranne MH, Michelle AD, et al. Effects of rhein on human articular chondrocytes in alginate beads. Bioche Pharmacology.2003,65(3):377-388.
[116].Wu SQ, Otero M, Unger FM, et al. Anti-inflammatory activity of an ethanolic Caesalpinia sappan extract in human chondrocytes and macrophagesJournal of Ethnopharmacology.2011,138(2):364-372.
[117].Sanchez C, Mathy-Hartert M, Deberg MA, et al. Effects of rhein on human articular chondrocytes in alginate beads. Biochem Pharmacol.2003,65(3):377-388.
[118].Kusuzaki K, Sugimoto S, Takeshita H, et al. DNA cytofluorometric analysis of chondrocytes in human articular cartilages under normal aging or arthritic conditions. Osteoarthritis Cartilage.2001,9(7):664-670.
[119].Duman-Scheel M, Weng L, Xin S, et al. Hedgehog regulates cell growth and proliferation by inducing cyclin D and cyclin E. Nature.2002,417(6886):299-304.
[120].Cossarizza A, Baccarani-Contri M, Kalashnikova G, et al. A new method for the cytofluorimetric analysis of mitochondrial membrane potential using the J-aggregate forming lipophilic cation 5,5',6,6'-tetrachloro-1,1',3,3' tetraeth-ylbenzimidazolylcarbocyanine iodide (JC-1).Biochem Biophys Res Commun.1993,197(1):40-45.
[121].Smiley ST, Reers M, Mottola-Hartshom C, et al. Intracellular heterogeneity in mitochondrial membrane potentials revealed by a J-aggregate forming lipophilic cation JC-1. Proc Natl Acad Sci USA.1991,88(9):3671-3675.
[1].王爱武,刘娅,雒琪,等.独活寄生汤抗炎、镇痛作用的药效学研究[J].中国实验方剂学杂志,2008,14(12)：61-64.
[2].赵玉珍,傅正宗,卢春凤,等.复方独活寄生合剂抗炎、镇痛作用观察[J].黑龙江医药科学,2003,26(3)：3-4.
[3].方剑乔,刘金洪,赵天征,等.独活寄生汤对小鼠胶原性关节炎治疗作用的研究[J].中国中医药科技,2000,7(5)：289-290.
[4].刘英杰,马利杰,王相利.独活寄生汤对兔膝骨性关节炎关节液白细胞介素-1和肿瘤坏死因子的影响[J].河北中医,2007,29(8)：748-750.
[5],李爱萍,何昌谋.独活寄生汤对兔膝骨关节炎体液中NO、SOD水平的影响[J].陕西中医,2010,31(10)：1430-1431.
[6].程维,张玉辉.独活寄生汤对膝骨性关节炎患者关节滑液中细胞因子的影响[J].辽宁中医药大学学报,2011,13(10)：216-217.
[7].朱自平.独活寄生汤对微循环的影响[J].中成药,1991,13(3)：26.
[8].许青嫒,谢人明,任军鹏等.独活寄生汤对麻醉动物脑循环的作用[J].陕西中医,1989,10(9)：425.
[9].陈筑红,李悦,杨凡,等.独活寄生汤对激素性股骨头缺血坏死的治疗作用[J].现代中西医结合杂志,2006,15(11)：1493.
[10].张若楠,王三虎,任东青.独活寄生汤对荷瘤小鼠的抗肿瘤作用研究[J].中国实验方剂学杂志,2007,13(10)：28-31.
[11].段泾云.独活寄生汤抗炎免疫药理作用研究[J].中成药研究,1988,(5)：28.
[12].王爱武,刘娅,林晓燕,等.独活寄生汤及其配方颗粒对胶原诱导型关节炎大鼠滑膜组织病理学变化的影响[J].中国中医药科技,2010,17(3)：202-203.
[13]].孙学东,姚华,姚杰.独活寄生汤加减治疗膝骨关节炎[J].北京中医药,2008,27(5)：362-363.
[14].李宝春.独活寄生汤加减治疗膝关节骨性关节炎100例临床观察[J].中医中药,2010,17(15)：80-81.
[15]].周友连.独活寄生汤加减治疗膝关节骨性关节炎116例疗效观察[J].浙江中医杂志,2007,42(10)：591.
[16].余建华,张衡.独活寄生汤治疗膝骨关节炎临床观察[J].中国实验方剂学杂志,2010,16(7)：215-217.
[17].高云.独活寄生汤加减治疗膝关节骨性关节炎临床疗效分析[J].中医中药,2010,7(20)：73.
[18].岑祖怡,周剑鹏.独活寄生汤治疗膝关节骨性关节炎50例疗效观察[J].新中医,2010,42(8)：50-51.
[19]].顾玉潜.独活寄生汤治疗膝关节骨性关节炎60例疗效观察[J].长春中医药大学学报,2008,24(2)：202-203.
[20].朱健儿.独活寄生汤治疗退行性膝骨关节炎60例[J].浙江中医学院学报,2005,29(4)：40.
[21].李培豪.独活寄生汤配合玻璃酸钠关节腔注射治疗膝骨性关节炎86例[J].中国中医急症,2010,19(7)：1236.
[22].管亦坚.关节腔内注射配合独活寄生汤治疗膝骨关节炎滑膜积液疗效观察[J].浙江中医药大学学报,2010,34(3)：399.
[23].于泽来,王伟,林一峰.独活寄生汤配合局部手法理伤治疗膝关节退行性骨关节炎[J].中国临床康复,2005,9(38)：124-125.
[24].邓泽前,张荣龙,俞宙.中药配合理疗手法治疗膝关节骨性关节炎106例分析[J].中国误诊学杂志,2008,8(34)：8466-8467.
[25].江飞,张清文,贺飞,等.独活寄生汤配合针刺治疗膝关节骨性炎症[J].中华全科医学,2010,8(9)：1156-1157.
[26].杨梦林,华刚,刘守新.中药加针灸治疗膝关节骨性关节炎164例[J].吉林中医药,2007,27(2)：41.
[27].汪利合.独活寄生汤加减结合运动疗法治疗膝骨关节炎的临床研究[J].中医正骨,2011,23(9)：3-5.
[28].刘俊峰,张卫华.独活寄生汤加减配合中药薰蒸治膝骨性关节炎50例[J].江西中医药,2011,42(4)：42-44.
[29].王武炼,叶锦霞,刘献祥.独活寄生汤加减内服外洗治疗膝骨性关节炎66例临床观察[J].福建中医药大学学报,2011,21(2)：44-45.
[30].李振荣,吕长青.独活寄生汤加减内服外用治疗膝骨关节炎36例[J].陕西中医,2011,32(9)：1168-1169.
[31].徐新玉,包力,韩明霞,等.独活寄生汤合中药离子导入疗膝关节骨性关节炎83例[J].中国中医药科技,2005,12(4)：259.
[32].杨芳,王月琴.独活寄生汤加减配合中药雾化治疗骨性关节炎163例[J].现代中医药,2007,27(5)：23-24.
[33].钟利进,陈懋生.独活寄生汤联合维生素D3治疗膝关节骨性关节炎的疗效[J].中医药导报,2009,15(9)：22-23.
[34].刘志文,赵淑平,陈茵,等.天灸联合独活寄生汤治疗膝骨关节炎96例[J].广东医学,2010,31(13)：1754-1755.
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