温经通络法对类风湿关节炎滑膜血管新生的干预作用研究
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摘要
类风湿关节炎(rheumatoid arthritis, RA)是一种以关节滑膜炎症为特征的慢性全身性自身免疫性疾病,滑膜炎可反复发作,导致关节软骨及骨质破坏,最终导致关节畸形及功能障碍。RA病理过程中,血管新生起着重要作用,且其出现较早,甚至在炎症特异的临床和组织学征象出现前就已经发生,最终造成滑膜炎、血管翳生长、骨和软骨破坏及骨赘形成。络病即是以络脉阻滞为特点的一类病证。“络病”理论认为络病的发生发展与血管病变密切相关。临床上从络辨治痹证一般采用温通络脉,活血通络为常用治则。导师朱跃兰教授在临床工作中治疗类风湿关节炎患者时发现,应用芍药甘草附子汤加味对改善RA患者临床症状和相关炎性指标、血液流变学等方面有一定的作用,该中药复方对于类风湿关节炎改善作用可能与影响滑膜血管异常增生相关。
一、综述
从中医和西医两个方面进行论述,中医方面对从络治痹的病因病机、临床表现特点及治疗等方面进行探讨；西医方面对VEGF及其受体和RA的相关性做一简要概述。
二、临床研究
目的：观察芍药甘草附子汤对RA患者的临床疗效以及对患者血清血管内皮生长因子VEGF的影响。
方法：采用随机对照的方法,将72例中医辨证为寒湿痹阻证RA患者按照随机区组设计(每组4人),随机分成两组,治疗组(芍药甘草附子汤组)36例和对照组(雷公藤多甙片组)36例。采用中医证候疗效判定标准以及国际公认的DAS28标准、ACR反应标准进行疗效评价,并对与风湿病相关的实验室指标及VEGF进行统计分析。
结果：入组72例患者中3例因依从性差退出实验,治疗组1例,对照组2例。两组治疗前基线水平无偏差。两组治疗前后中医疗效观察及症状积分比较,两组均没有发现显著疗效患者,其中治疗组35例,有效25例,占71.43%；10例无效,占28.57%。对照组34例,23例有效,占67.65%；11例无效,占32.35%。两组患者每组治疗前后中医症状积分经配对样本t检验,P<0.01,有显著统计学差异。两组组间在同一时间点比较,P>0.05,无统计学意义。两组在治疗后4周DAS28评分较基线相比较无明显统计学差异(P>0.05),8周后与4周后、12周后较8周后相比也均无明显统计学差异(P>0.05)。治疗组治疗后8周DAS28评分较基线相比较下降有明显统计学意义(P<0.05),12周时DAS28评分较基线和4周时相比较下降均有显著统计学意义(P<0.01)。对照组治疗后8周DAS28评分较基线相比较以及12周时DAS28评分较4周时相比较下降有明显统计学意义(P<0.05),12周时较基线相比较下降有显著统计学意义(P<0.01)。两组组间在同一时间点上的DAS28评分无明显统计学差异(P>0.05)。疗程结束后,治疗组总有效率68.57%,对照组总有效率64.71%。经Ridit分析,治疗组R11=0.492,对照组R2=0.508,u=—0.237,P>0.05,两组比较无统计学差异。两组患者治疗4周、8周、12周后ACR20达标率在同一时间点比较经卡方检验,P>0.05,无统计学显著性差异。治疗结束后即12周末治疗组达到ACR20患者26例,占该组总人数的74.29%；对照组达到ACR20患者24例,占该组总人数的70.59%。两组在治疗8周和12周后ACR50达标率在同一时间点比较经卡方检验,P>0.05,无统计学显著性差异。治疗12周后达到ACR50患者治疗组8例,占22.86%；对照组6例,占17.65%。两组在治疗8周和12周后均未出现ACR70达标患者。实验室指标观察结果显示,两组在治疗干预后各项指标均呈下降趋势。经方差分析,4周后除对照组CRP与治疗前相比下降不显著外(P>0.05),其余均较治疗前明显下降(P<0.05),8周、12周后ESR、CRP与治疗前相比均有显著下降(P<0.01)。两组组间同一时间点相比较无显著差异(P>0.05)。两组患者血清VEGF在治疗12周后较治疗前显著降低(P<0.01),两组组间同一时间点相比较差异不显著(P>0.05)。安全性指标检测结果,两组治疗前后血、尿、便常规均未见明显异常。治疗过程中对照组有1例出现ALT升高,给予葡醛内酯治疗2周后恢复正常,其余未见明显异常。整体前后对照未见明显差异。
结论：应用芍药甘草附子汤加味治疗寒湿痹阻证RA患者临床疗效满意。实验室指标观察结果显示,RA患者血清VEGF在治疗前有不同程度的升高,治疗后也都得到了不同程度的降低。
三、实验研究
目的：观察CIA大鼠血清和滑膜组织中VEGF及其受体VEGFR-2的表达变化,以及芍药甘草附子汤对其干预作用影响。
方法：选用SPF级Wistar大鼠,采用随机数字表法分为正常组、模型组、治疗组(芍药甘草附子汤组)和对照组(雷公藤多甙片组)。药物干预后进行形态学观察并计算AI、X-ray观察等。药物干预结束后采用ELISA法、免疫组织化学法以及realtime PCR法检测VEGF、sVEGFR-2/VEGFR-2在大鼠血清和滑膜里的表达水平。
结果：①造模后各组大鼠表现出程度不等的炎症反应、足趾关节肿胀,体重也较正常组增长减慢。用药后各组大鼠炎症反应明显好转,对照组和治疗组明显好于模型组(P<0.05),但两组之间无显著差异(P>0.05)。大鼠关节病理评分：正常组双侧踝关节组织学结构正常；模型组滑膜可见程度不等的炎性细胞浸润、水肿、肉芽组织形成,甚至出现较明显的关节软骨破坏；各用药组光镜下病理改变较模型组减轻,炎细胞浸润、滑膜增生减轻,软骨破坏程度减弱。大鼠X-ray表现：CIA大鼠X线表现是以膝关节、踝关节及趾间关节受累为主。呈对称性,具有明显的骨皮质变薄、骨质疏松表现。边缘性骨侵蚀,伴有骨质增生性反应,部分关节骨破坏明显。骨端增生、粗大,关节间隙狭窄或增宽甚至消失。②采用ELISA法检测血清VEGF、VEGFR-2含量,模型组明显高于正常组(P<0.05),用药后对照组和治疗组明显下降(P<0.05),对照组VEGF表达水平略低于治疗组,二者差异无统计学意义(P>0.05)。VEGF、VEGFR-2免疫组化标记均定位于细胞浆,以细胞浆着色呈棕黄色为阳性细胞。正常组大鼠滑膜细胞的胞浆上可见少量均匀分布的棕黄色颗粒；模型组滑膜细胞的胞浆上的棕黄色颗粒明显多而密集,较正常组有明显差异(P<0.05)；对照组和治疗组的阳性细胞数减少,与模型组相比有显著性差异(P<0.05)。③结果经统计学分析,模型组VEGF、 VEGFR-2表达水平与正常组相比较明显偏高(P<0.05),治疗组和对照组VEGF、 VEGFR-2表达水平与模型组相比较下降明显(P<0.05),但两组之间无明显差异(P>0.05)。
结论：芍药甘草附子汤能明显降低CIA大鼠关节炎症指数、减轻足趾肿胀程度及踝关节病理评分；能够降低CIA大鼠血清中VEGF及其受体的水平,明显下调滑膜组织VEGF、VEGFR-2的表达；能够明显抑制CIA大鼠滑膜组织VEGFmRNA、VEGFR-2mRNA的表达,其作用的具体机制尚待进一步研究。
Rheumatoid Arthritis (RA) is an autoimmune disease characterized by synovial hyperplasia and joint deformity. Angiogenesis plays an important role in the process of RA. RA pathology, in the process of angiogenesis plays an important role, and it appears early, even in the presence of specific clinical and histological signs of inflammation, has happened before, resulting in synovitis, pannus growth, bone and cartilage destruction, and osteophyte formation. Block for the characteristics of disease is that Luo Bing."Luo Bing" theory means that the disease are closely associated with vascular lesions. From clinical syndrome differentiation treatment of commonly used WenTong collaterals, activating blood know luo for commonly used therapeutic principle. Mentor professor Zhu Yuelan treatment of rheumatoid arthritis patients in the clinical work, application of peony liquorice monkshood soup taste for improving the clinical symptoms of patients with RA and associated inflammatory indexes, blood rheology have certain effect, to improve the compound Chinese medicine for rheumatoid arthritis may affect synovial hyperplasia of vascular abnormalities related.
REVIEW
First, we discuss the etiology, pathogenesis, clinical character-ristics and treatment of Chinese medicine of treating Bi Zheng using Tong Luo.Then,we discuss the correlation of VEGF with RA.
CLINICAL RESEARCH
OBJECTIVE:To study the clinical effect of Shaoyao-Gancao-Fuzi soup of RA patients and the effect on vascular endothelial growth factor (VEGF) of the patients serum.
METHODS:Using randomized controlled methods,60RA cases which of ShenQi XuHan syndrome were divided into treatment group and control group.30cases of treatment group which treated by Shaoyao-Gancao-Fuzi soap and30cases of control group which treated by Tripterygium Glycosides. To evaluate curative effect by using TCM syndrome judgment standard and DAS28curative effect evaluation standard, the ACR reaction, and the laboratory indicators including VEGF associated with rheumatism were statistically analyzed.
RESULTS:72patients in3cases due to poor adherence exit the experiment,1 case in the treatment group,2cases in the control group.The two groups'baseline have no deviation before treatment(P>0.05).Patients of the two groups both have no significant curative effect evaluated by TCM symptoms observation and integral comparison before and after the treatment.Effective rate of treatment group is71.43%, this rate in the control is67.65%.No significant difference in total effective rate was found between the two groups(P>0.05).Accord to standard of DAS28,there is no obvious change compared with baseline in the two groups after4weeks treatment (P>0.05),Same as8weeks treatment compared with4weeks,12weeks treatment compared with8weeks. After8weeks treatment,treatment group has significantly decreased (P<0.05), and has declined significantly after12weeks treatment(P<0.01). Control group also has significantly decreased compared to a baseline after8weeks treatment (P<0.05),and has declined significantly after12weeks treatment(P <0.01). No significant difference was found between the two groups at the same point.(P>0.05). After the treatment, the total effective rate of treatment group is68.57%, this rate in the control is64.71%. The Ridit analysis show that R1=0.492(treatment group), R2=0.508(control group), u=-0.237,P>0.05.There is no significant difference between two groups. There is no statistically significant difference between two groups according to ACR20standard at the same time point(P>0.05).After treatments, number of cases reaching ACR20is26which accounting for74.29%of the total number in this group.The number in control group is24which accounting for70.59%of the total number of the group. The number of cases reaching ACR50has no statistically significant difference in8weeks and12weeks after treatment at the same time point (P>0.05).There is no cases reaching ACR70after the treatment of8weeks and12weeks.Laboratory results showed ESR, CRP and VEGF reduced significantly than baseline data in both groups(P<0.01). There is no statistically significant difference between two groups at the same point in time(P>0.05). Safety index test results showed that there is no obvious abnormal in the blood, urine and stool.There is1case which appeared high ALT,and soon returned to normal after2weeks treatment.The rest has no obvious anomalies. Overall control did not see obvious difference before and after.
CONCLUSION:ShaoYao-GanCao-FuZi soap has good clinical curative effect in the RA patient of ShenQiXuHan type. Laboratory observation results show that RA patients serum VEGF is higher before treatment, and is droped after treatment.
EXPERIMENTAL RESEARCH
OBJECTIVE:To study expression of VEGF and its receptor VEGFR-2in the CIA rats serum and synovial tissue, and the effect of ShaoYao-GanCao-FuZi soap on their expressioins.
METHODS:SPF level Wistar rats were randomly divided into normal group、 model group、treatment group (treated by ShaoYao-GanCao-FuZi soup) and control group (treated by Tripterygium Glycosides). Arthritis index and X-ray abnormalities were evaluated by two independent observers blinded to the groups of the animals. The level of VEGF and sVEGFR-2/VEGFR-2in serum and synovium were studied by ELISA、immunohistochemical staining and realtime PCR respectively.
RESULTS:①The CIA model rats showed varying degrees of joint inflammation, swelling of the toes and joint, slow growth of the weight compared with the normal group. Degrees of inflammation droped apparently after treatment (P <0.05),but there was no significant difference between two groups (P>0.05).Rats joint pathology score show that normal histological structure in the normal bilateral ankle group.Inflammatory cell infiltration, edema, granulation tissue formation and even more apparent destruction of articular cartilage showed in the synovial of Model group.All about it would decreased after treatmend. X-ray of rats showed changes in the knee joint, ankle and toe joints. Cortical thinning of bone, osteoporosis,Marginal bone erosion, bone proliferative reaction, joint bone destruction can be seen. Even bone hyperplasia, bulky, joint space narrowing or widening disappear.②Using the ELISA method to detect VEGF and VEGFR-2of the serum, the model group was obviously higher than normal group (P<0.05).After the treatment, control group and treatment group significantly decreased (P<0.05). Expression of VEGF in the control group is slightly lower than the treatment group, and there was no statistically significant difference between them (P>0.05).VEGF and VEGFR-2are located in the cytoplasm and cytoplasmic staining for a tan for positive cells by the immune histochemical.Synovial cell cytoplasm of normal group rats is visible on a small amount of evenly distributed brownish-yellow granules.Synovial cell cytoplasm brownish-yellow granules is significantly more dense on model group which is obvious differences compare with the normal group (P<0.05).The number of positive cells decreased in the both Control group and treatment group which are significant difference compared with the model group (P<0.05).③By statistics analysis,the expression of VEGF and VEGFR-2on the model group is significantly higher than the normal group (P<0.05).The expression of VEGF and VEGFR-2on the treatment group and control group is decreased significantly compared with the model group (P <0.05), but there is no significant difference between two groups (P>0.05).
CONCLUSION:ShaoYao-GanCao-FuZi soup can obviously reduce the CIA rat' articular inflammation index, reduce its swelling degree、pathological grading、levels of VEGF and its receptor on serum and expression of VEGF and VEGFR-2on synovial tissue. The role of the specific mechanism remains to be further research.
一、综述
从中医和西医两个方面进行论述,中医方面对从络治痹的病因病机、临床表现特点及治疗等方面进行探讨；西医方面对VEGF及其受体和RA的相关性做一简要概述。
二、临床研究
目的：观察芍药甘草附子汤对RA患者的临床疗效以及对患者血清血管内皮生长因子VEGF的影响。
方法：采用随机对照的方法,将72例中医辨证为寒湿痹阻证RA患者按照随机区组设计(每组4人),随机分成两组,治疗组(芍药甘草附子汤组)36例和对照组(雷公藤多甙片组)36例。采用中医证候疗效判定标准以及国际公认的DAS28标准、ACR反应标准进行疗效评价,并对与风湿病相关的实验室指标及VEGF进行统计分析。
结果：入组72例患者中3例因依从性差退出实验,治疗组1例,对照组2例。两组治疗前基线水平无偏差。两组治疗前后中医疗效观察及症状积分比较,两组均没有发现显著疗效患者,其中治疗组35例,有效25例,占71.43%；10例无效,占28.57%。对照组34例,23例有效,占67.65%；11例无效,占32.35%。两组患者每组治疗前后中医症状积分经配对样本t检验,P<0.01,有显著统计学差异。两组组间在同一时间点比较,P>0.05,无统计学意义。两组在治疗后4周DAS28评分较基线相比较无明显统计学差异(P>0.05),8周后与4周后、12周后较8周后相比也均无明显统计学差异(P>0.05)。治疗组治疗后8周DAS28评分较基线相比较下降有明显统计学意义(P<0.05),12周时DAS28评分较基线和4周时相比较下降均有显著统计学意义(P<0.01)。对照组治疗后8周DAS28评分较基线相比较以及12周时DAS28评分较4周时相比较下降有明显统计学意义(P<0.05),12周时较基线相比较下降有显著统计学意义(P<0.01)。两组组间在同一时间点上的DAS28评分无明显统计学差异(P>0.05)。疗程结束后,治疗组总有效率68.57%,对照组总有效率64.71%。经Ridit分析,治疗组R11=0.492,对照组R2=0.508,u=—0.237,P>0.05,两组比较无统计学差异。两组患者治疗4周、8周、12周后ACR20达标率在同一时间点比较经卡方检验,P>0.05,无统计学显著性差异。治疗结束后即12周末治疗组达到ACR20患者26例,占该组总人数的74.29%；对照组达到ACR20患者24例,占该组总人数的70.59%。两组在治疗8周和12周后ACR50达标率在同一时间点比较经卡方检验,P>0.05,无统计学显著性差异。治疗12周后达到ACR50患者治疗组8例,占22.86%；对照组6例,占17.65%。两组在治疗8周和12周后均未出现ACR70达标患者。实验室指标观察结果显示,两组在治疗干预后各项指标均呈下降趋势。经方差分析,4周后除对照组CRP与治疗前相比下降不显著外(P>0.05),其余均较治疗前明显下降(P<0.05),8周、12周后ESR、CRP与治疗前相比均有显著下降(P<0.01)。两组组间同一时间点相比较无显著差异(P>0.05)。两组患者血清VEGF在治疗12周后较治疗前显著降低(P<0.01),两组组间同一时间点相比较差异不显著(P>0.05)。安全性指标检测结果,两组治疗前后血、尿、便常规均未见明显异常。治疗过程中对照组有1例出现ALT升高,给予葡醛内酯治疗2周后恢复正常,其余未见明显异常。整体前后对照未见明显差异。
结论：应用芍药甘草附子汤加味治疗寒湿痹阻证RA患者临床疗效满意。实验室指标观察结果显示,RA患者血清VEGF在治疗前有不同程度的升高,治疗后也都得到了不同程度的降低。
三、实验研究
目的：观察CIA大鼠血清和滑膜组织中VEGF及其受体VEGFR-2的表达变化,以及芍药甘草附子汤对其干预作用影响。
方法：选用SPF级Wistar大鼠,采用随机数字表法分为正常组、模型组、治疗组(芍药甘草附子汤组)和对照组(雷公藤多甙片组)。药物干预后进行形态学观察并计算AI、X-ray观察等。药物干预结束后采用ELISA法、免疫组织化学法以及realtime PCR法检测VEGF、sVEGFR-2/VEGFR-2在大鼠血清和滑膜里的表达水平。
结果：①造模后各组大鼠表现出程度不等的炎症反应、足趾关节肿胀,体重也较正常组增长减慢。用药后各组大鼠炎症反应明显好转,对照组和治疗组明显好于模型组(P<0.05),但两组之间无显著差异(P>0.05)。大鼠关节病理评分：正常组双侧踝关节组织学结构正常；模型组滑膜可见程度不等的炎性细胞浸润、水肿、肉芽组织形成,甚至出现较明显的关节软骨破坏；各用药组光镜下病理改变较模型组减轻,炎细胞浸润、滑膜增生减轻,软骨破坏程度减弱。大鼠X-ray表现：CIA大鼠X线表现是以膝关节、踝关节及趾间关节受累为主。呈对称性,具有明显的骨皮质变薄、骨质疏松表现。边缘性骨侵蚀,伴有骨质增生性反应,部分关节骨破坏明显。骨端增生、粗大,关节间隙狭窄或增宽甚至消失。②采用ELISA法检测血清VEGF、VEGFR-2含量,模型组明显高于正常组(P<0.05),用药后对照组和治疗组明显下降(P<0.05),对照组VEGF表达水平略低于治疗组,二者差异无统计学意义(P>0.05)。VEGF、VEGFR-2免疫组化标记均定位于细胞浆,以细胞浆着色呈棕黄色为阳性细胞。正常组大鼠滑膜细胞的胞浆上可见少量均匀分布的棕黄色颗粒；模型组滑膜细胞的胞浆上的棕黄色颗粒明显多而密集,较正常组有明显差异(P<0.05)；对照组和治疗组的阳性细胞数减少,与模型组相比有显著性差异(P<0.05)。③结果经统计学分析,模型组VEGF、 VEGFR-2表达水平与正常组相比较明显偏高(P<0.05),治疗组和对照组VEGF、 VEGFR-2表达水平与模型组相比较下降明显(P<0.05),但两组之间无明显差异(P>0.05)。
结论：芍药甘草附子汤能明显降低CIA大鼠关节炎症指数、减轻足趾肿胀程度及踝关节病理评分；能够降低CIA大鼠血清中VEGF及其受体的水平,明显下调滑膜组织VEGF、VEGFR-2的表达；能够明显抑制CIA大鼠滑膜组织VEGFmRNA、VEGFR-2mRNA的表达,其作用的具体机制尚待进一步研究。
Rheumatoid Arthritis (RA) is an autoimmune disease characterized by synovial hyperplasia and joint deformity. Angiogenesis plays an important role in the process of RA. RA pathology, in the process of angiogenesis plays an important role, and it appears early, even in the presence of specific clinical and histological signs of inflammation, has happened before, resulting in synovitis, pannus growth, bone and cartilage destruction, and osteophyte formation. Block for the characteristics of disease is that Luo Bing."Luo Bing" theory means that the disease are closely associated with vascular lesions. From clinical syndrome differentiation treatment of commonly used WenTong collaterals, activating blood know luo for commonly used therapeutic principle. Mentor professor Zhu Yuelan treatment of rheumatoid arthritis patients in the clinical work, application of peony liquorice monkshood soup taste for improving the clinical symptoms of patients with RA and associated inflammatory indexes, blood rheology have certain effect, to improve the compound Chinese medicine for rheumatoid arthritis may affect synovial hyperplasia of vascular abnormalities related.
REVIEW
First, we discuss the etiology, pathogenesis, clinical character-ristics and treatment of Chinese medicine of treating Bi Zheng using Tong Luo.Then,we discuss the correlation of VEGF with RA.
CLINICAL RESEARCH
OBJECTIVE:To study the clinical effect of Shaoyao-Gancao-Fuzi soup of RA patients and the effect on vascular endothelial growth factor (VEGF) of the patients serum.
METHODS:Using randomized controlled methods,60RA cases which of ShenQi XuHan syndrome were divided into treatment group and control group.30cases of treatment group which treated by Shaoyao-Gancao-Fuzi soap and30cases of control group which treated by Tripterygium Glycosides. To evaluate curative effect by using TCM syndrome judgment standard and DAS28curative effect evaluation standard, the ACR reaction, and the laboratory indicators including VEGF associated with rheumatism were statistically analyzed.
RESULTS:72patients in3cases due to poor adherence exit the experiment,1 case in the treatment group,2cases in the control group.The two groups'baseline have no deviation before treatment(P>0.05).Patients of the two groups both have no significant curative effect evaluated by TCM symptoms observation and integral comparison before and after the treatment.Effective rate of treatment group is71.43%, this rate in the control is67.65%.No significant difference in total effective rate was found between the two groups(P>0.05).Accord to standard of DAS28,there is no obvious change compared with baseline in the two groups after4weeks treatment (P>0.05),Same as8weeks treatment compared with4weeks,12weeks treatment compared with8weeks. After8weeks treatment,treatment group has significantly decreased (P<0.05), and has declined significantly after12weeks treatment(P<0.01). Control group also has significantly decreased compared to a baseline after8weeks treatment (P<0.05),and has declined significantly after12weeks treatment(P <0.01). No significant difference was found between the two groups at the same point.(P>0.05). After the treatment, the total effective rate of treatment group is68.57%, this rate in the control is64.71%. The Ridit analysis show that R1=0.492(treatment group), R2=0.508(control group), u=-0.237,P>0.05.There is no significant difference between two groups. There is no statistically significant difference between two groups according to ACR20standard at the same time point(P>0.05).After treatments, number of cases reaching ACR20is26which accounting for74.29%of the total number in this group.The number in control group is24which accounting for70.59%of the total number of the group. The number of cases reaching ACR50has no statistically significant difference in8weeks and12weeks after treatment at the same time point (P>0.05).There is no cases reaching ACR70after the treatment of8weeks and12weeks.Laboratory results showed ESR, CRP and VEGF reduced significantly than baseline data in both groups(P<0.01). There is no statistically significant difference between two groups at the same point in time(P>0.05). Safety index test results showed that there is no obvious abnormal in the blood, urine and stool.There is1case which appeared high ALT,and soon returned to normal after2weeks treatment.The rest has no obvious anomalies. Overall control did not see obvious difference before and after.
CONCLUSION:ShaoYao-GanCao-FuZi soap has good clinical curative effect in the RA patient of ShenQiXuHan type. Laboratory observation results show that RA patients serum VEGF is higher before treatment, and is droped after treatment.
EXPERIMENTAL RESEARCH
OBJECTIVE:To study expression of VEGF and its receptor VEGFR-2in the CIA rats serum and synovial tissue, and the effect of ShaoYao-GanCao-FuZi soap on their expressioins.
METHODS:SPF level Wistar rats were randomly divided into normal group、 model group、treatment group (treated by ShaoYao-GanCao-FuZi soup) and control group (treated by Tripterygium Glycosides). Arthritis index and X-ray abnormalities were evaluated by two independent observers blinded to the groups of the animals. The level of VEGF and sVEGFR-2/VEGFR-2in serum and synovium were studied by ELISA、immunohistochemical staining and realtime PCR respectively.
RESULTS:①The CIA model rats showed varying degrees of joint inflammation, swelling of the toes and joint, slow growth of the weight compared with the normal group. Degrees of inflammation droped apparently after treatment (P <0.05),but there was no significant difference between two groups (P>0.05).Rats joint pathology score show that normal histological structure in the normal bilateral ankle group.Inflammatory cell infiltration, edema, granulation tissue formation and even more apparent destruction of articular cartilage showed in the synovial of Model group.All about it would decreased after treatmend. X-ray of rats showed changes in the knee joint, ankle and toe joints. Cortical thinning of bone, osteoporosis,Marginal bone erosion, bone proliferative reaction, joint bone destruction can be seen. Even bone hyperplasia, bulky, joint space narrowing or widening disappear.②Using the ELISA method to detect VEGF and VEGFR-2of the serum, the model group was obviously higher than normal group (P<0.05).After the treatment, control group and treatment group significantly decreased (P<0.05). Expression of VEGF in the control group is slightly lower than the treatment group, and there was no statistically significant difference between them (P>0.05).VEGF and VEGFR-2are located in the cytoplasm and cytoplasmic staining for a tan for positive cells by the immune histochemical.Synovial cell cytoplasm of normal group rats is visible on a small amount of evenly distributed brownish-yellow granules.Synovial cell cytoplasm brownish-yellow granules is significantly more dense on model group which is obvious differences compare with the normal group (P<0.05).The number of positive cells decreased in the both Control group and treatment group which are significant difference compared with the model group (P<0.05).③By statistics analysis,the expression of VEGF and VEGFR-2on the model group is significantly higher than the normal group (P<0.05).The expression of VEGF and VEGFR-2on the treatment group and control group is decreased significantly compared with the model group (P <0.05), but there is no significant difference between two groups (P>0.05).
CONCLUSION:ShaoYao-GanCao-FuZi soup can obviously reduce the CIA rat' articular inflammation index, reduce its swelling degree、pathological grading、levels of VEGF and its receptor on serum and expression of VEGF and VEGFR-2on synovial tissue. The role of the specific mechanism remains to be further research.
引文
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