摘要
目的:探讨雪莲强筋壮骨方对膝骨关节炎(knee osteoarthritis,KOA)软骨细胞p38丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)信号传导通路的影响。方法:将24只3月龄雄性新西兰大白兔随机分为空白组、模型组、p38阻断剂组及含药血清组,每组6只。除空白组外,其余各组均采用改良Hulth法行KOA造模。造模成功后含药血清组以雪莲强筋壮骨方水煎剂灌胃(10 g·kg^(-1)),其余各组均以等量生理盐水灌胃,每天2次,连续干预7 d后静脉采血,分别制成10%含药血清和10%空白血清的DMEM培养液保存备用。血清制备完成后处死动物,分离膝关节软骨进行软骨细胞培养。空白组和模型组以空白血清进行干预,p38阻断剂组以空白血清和SB203580进行干预,含药血清组以含药血清进行干预。血清干预结束后,分别采用免疫荧光技术和蛋白免疫印迹法检测各组软骨细胞磷酸化p38MAPK水平。结果:免疫荧光检测结果显示,模型组磷酸化p38MAPK表达呈强阳性,p38阻断剂组和含药血清组次之,空白组最弱。蛋白免疫印迹检测结果显示,空白组磷酸化p38MAPK表达最弱,模型组磷酸化p38MAPK表达最强,p38阻断剂组和含药血清组磷酸化p38MAPK表达水平相当,介于空白组和模型组之间。经扫描定量检测,空白组、模型组、p38阻断剂组和含药血清组条带的光密度值分别为0.31±0.06、0.83±0.11、0.58±0.45、0.55±0.56,差异有统计学意义(F=3.872,P=0.034)。空白组的光密度值小于模型组、p38阻断剂组及含药血清组(P=0.007,P=0.031,P=0.038);模型组的光密度值大于p38阻断剂组和含药血清组(P=0.038,P=0.028);p38阻断剂组和含药血清组的光密度值比较,差异无统计学意义(P=0.066)。结论:雪莲强筋壮骨方能阻滞KOA软骨细胞p38MAPK信号传导通路,其效果与p38MAPK阻断剂SB203580相当。
Objective ：To explore the effects of Xuelian Qiangjin Zhuanggu Fang（ 雪莲强筋壮骨方 ,XLQJZGF）on p38 mitogen-activated protein kinase （MAPK） signaling transduction pathway in chondrocytes of patients with knee osteoarlhritis （KOA）. Methods ：Twenly four three-month-old male New Zealand rabbits were randomly divided into blank group, model group, p38 blocker group and medicated serum group,6 eases in each group. The KOA rabbit models were created in model group,p38 blocker group and medicated serum group by using modified Hulth method. After successful modeling, the rabbits in medicated serum group were intragastric administrated with XLQJZGF decoction（ 10 g/kg） ,while the rabbits in blank group,model group and p38 blocker group were intragastric administrated with isodose normal saline ,twice a day for consecutive 7 days. Then their blood were fetched out from veins for making DMEM culture fluid added with 10% medicated serum and DMEM culture fluid added with 10% blank serum respectively for future use. After successful serum preparation,the rabbits were executed, and their chondrocytes were isolated from knee articular cartilages and were cultured in vitro. The chondrocytes were intervened with blank serum（ blank group and model group） ,blank serum and SB203580（ p38 blocker group）and medicated serum（ medi- cated serum group）respectively. After the end of serum intervention,the levels of phosphorylated-p38 MAPK in chondrocytes were detected by using immunofluorescence technique and Western blot assay respectively. Results：The results of immunofluorescence assay showed that the phosphorylated-p38 MAPK exhibited high-to-low expression in model group,p38 blocker group and medicated serum group and blank group in turn. The results of Western blot assay showed that the phosphorylated-p38 MAPK exhibited low-to-high expression in blank group,p38 blocker group and medicated serum group and model group. The scanning and quantitative detection results showed that there was statistical difference in optical densityvalues of strips between blank group, model group, p38 blocker group and medicated serum group （0.31 ＋/- 0.06,0.83 ＋/- 0.11,0.58 ＋/- 0.45,0.55 ＋/- 0.56 ; F = 3. 872, P = 0. 034）. The optical density values were smaller in blank group compared to model group, p38 blocker group and medicated serum group （P = 0. 007, P = 0. 031, P = 0.038 ） , and were larger in model group compared to p38 blocker group and medicated serum group （P = 0. 038,P = 0.028 ） , and there was no statistical differences in optical density values between p38 blocker group and medicated serum group（P = 0. 066）. Conclusion：XLQJZGF can block the p38 MAPK signaling transduction pathway in chondrocytes in patients with KOA, and it is sinfilar to SB203580（ p38 MAPK blocker）in curative effects.
作者
赵永胜
李发东
杨新成
孟刚
郭鑫
赵辉
潘有龙
王晓乐
ZHAO Yongsheng;LI Fadong;YANG Xincheng;MENG Gang;GUO Xin;ZHAO Hui;PAN Youlong;WANG Xiaole(Changji Hospital of Traditional Chinese Medicine,Changji 831100,Xinjiang,China)
出处
《中医正骨》
2018年第7期1-5,共5页
The Journal of Traditional Chinese Orthopedics and Traumatology
基金
新疆维吾尔自治区卫生计生委青年科技人才专项科研项目(2015Y08)
关键词
骨关节炎
雪莲强筋壮骨方
软骨细胞
38丝裂原活化蛋白激酶类
信号传导
osteoarthritis
Xuelian Qiangjin Zhuanggu Fang
ehondroeytes
p38 nfitogen - activated protein kinases
signal transduetion
