论新止骨增生丸通过抑制IL-1β对TGF-β/Smad信号通路下TGF-βR1、Smad2表达影响及作用机制研究  

Effect and Mechanism of Xinzhigu Zengsheng Pill(新止骨增生丸)on the Expression of TGF-β R1 and Smad2 in TGF-β/Smad Signaling Pathway by Inhibiting IL-1β

作　　者：马楠 金连峰[2] 武爽 MA Nan;JIN Lianfeng;WU Shuang(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,Liaoning,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847 [2]辽宁中医药大学附属医院,辽宁沈阳110032

出　　处：《辽宁中医药大学学报》2022年第3期22-26,共5页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81574050);辽宁省自然科学基金(201602482);沈阳市科技计划项目(18013083)。

摘　　要：目的利用细胞、分子生物学技术,采用蛋白免疫印迹(Western blot,WB)检测方法,观察新止骨增生丸对大鼠膝骨关节炎(knee osteoarthritis,KOA)软骨细胞的作用机制及对细胞凋亡的影响,并讨论新止骨增生丸治疗膝骨关节炎的作用机制。方法SPF级6月龄大鼠24只,随机分为两组,空白对照组6只,模型复制组18只。模型复制组大鼠采用前后交叉韧带离断术复制骨性关节炎大鼠模型,模型复制成功后,将该组大鼠再次随机分为3组:模型对照组(6只)、中药组(6只)及西药对照组(6只)。连续治疗6周后,取整个膝关节,分离软骨组织。膝关节病理切片HE染色法观察各组软骨细胞状态,采用WB法检测各组大鼠膝关节软骨组织中白细胞介素-1β(IL-1β)、转化生长因子β受体1(TGF-βR1)、白细胞抑制因子2(Smad2)蛋白表达,采用SPSS 16.0软件包对WB法检测结果进行方差分析,P<0.05为差异有统计学意义。结果(1)膝关节病理切片结果:经新止骨增生丸治疗后KOA症状减轻。(2)WB检测结果:模型对照组IL-1β、TGF-βR1、Smad2蛋白表达水平较空白对照组明显上调(均P<0.05);中药组、西药对照组IL-1β蛋白表达水平较模型对照组呈现明显下调(均P<0.05),且中药组变化最为显著;中药组、西药对照组TGF-βR1、Smad2蛋白表达水平较模型对照组呈现明显上调趋势(均P<0.05)。结论新止骨增生丸能够抑制IL-1β表达,激活TGF-β/Smad信号通路以上调TGF-βR1、Smad2蛋白表达,且中药组变化最为显著,达到治疗膝骨性关节炎的效果。Objective To observe the effect of Xinzhigu Zengsheng Pill(新止骨增生丸)on chondrocytes and apoptosis in rats with knee osteoarthritis(KOA)by Western blot(WB),and to discuss the mechanism of Xinzhigu Zengsheng Pill in the treatment of knee osteoarthritis.Methods Twenty four SPF grade 6-month-old rats were randomly divided into two groups,the blank group(n=6),18 rats in model replication group.The rats in the model replication group were treated with anterior and posterior cruciate ligament transection to reproduce the rat model of osteoarthritis.After the successful replication of the model,the reorganized rats were randomly divided into three groups:model group(n=6),traditional Chinese medicine group(n=6)and Western medicine group(n=6).After 6 weeks of continuous treatment,the whole knee joint was removed and the cartilage tissue was separated.HE staining was used to observe the state of chondrocytes in the pathological section of knee joint,and WB method was used to detect IL-1β,TGF-β R1,Smad2 protein in the cartilage tissue of knee joint,SPSS 16.0 software package was used to analyze the variance of WB method,P<0.05 with statistically significant.Results(1)Pathological section of knee joint results:KOA symptoms were relieved after treatment with Xinzhigu Zengsheng Pill.(2)Western Blot results:Compared with the blank control group,the expression levels of IL-1β,TGF-β R1 and Smad2 proteins in the model control group were significantly up-regulated(all P<0.05).Compared with model control group,the expression level of IL-1βprotein in TCM group and Western medicine control group was significantly down-regulated(P<0.05),and the change of IL-1β protein in TCM group was the most significant.Compared with model control group,the protein expression levels of TGF-βR1 and Smad2 in TCM group and Western medicine control group were significantly up-regulated(all P<0.05),and the change of TGF-β R1 and Smad2 protein in TCM group was the most significant.Conclusion Xinzhigu Zengsheng Pill can inhibit IL-1β expre

关 键 词：膝骨关节炎 新止骨增生丸 信号通路 IL-1Β TGF-ΒR1 SMAD2 

分 类 号：R684.3[医药卫生—骨科学]