扁蒴藤素调节YAP/TAZ信号通路对创伤性关节炎大鼠的治疗作用研究OACSTPCD
Study on Therapeutic Effects of Pristimerin on Post-Traumatic Osteoarthritis Rats by Regulating the YAP/TAZ Signaling Pathway
目的 探究扁蒴藤素(PM)调节Yes相关蛋白(YAP)/转录共激活因子PDZ结合基序(TAZ)信号通路对创伤性关节炎(PTOA)大鼠的治疗作用.方法 将SD雄性大鼠随机分为对照组、PTOA组、低剂量PM组(PM-L组,100 mg/kg PM)、高剂量PM组(PM-H组,200 mg/kg PM)和高剂量PM+YAP抑制剂VTPF组(PM-H+VTPF组,200 mg/kg PM+10 mg/kg VTPF),每组12 只.记录大鼠热痛阈值、压痛阈值的变化.HE染色观察大鼠软骨组织病理学变化,并进行Mankin's评分,酶联免疫吸附试验(ELISA)检测大鼠软骨组织中炎症因子一氧化氮(NO)、白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)和基质金属蛋白酶(MMP)-1、MMP-3 水平.TUNEL染色观察大鼠软骨组织细胞凋亡的情况.实时荧光定量PCR(RT-qPCR)实验检测大鼠软骨组织TAZ、IL-1β、TNF-α mRNA水平.Western Blot检测大鼠软骨组织YAP/TAZ信号通路相关蛋白的表达情况.结果 与对照组相比,PTOA组大鼠热痛阈值、压痛阈值、软骨组织YAP磷酸化水平、TAZ蛋白水平与mRNA水平显著降低(P<0.05),软骨组织IL-1β、TNF-α及其mRNA水平、NO、MMP-1、MMP3 水平、细胞凋亡率、Mankin's评分显著升高(P<0.05),HE显示软骨层变薄,结构不清晰,软骨细胞大量丢失、排列紊乱.与PTOA组相比,PM-L组、PM-H组大鼠相关指标变化与上述相反(P<0.05),软骨组织病变减轻.YAP抑制剂VTPF减轻了PM对PTOA的治疗作用.结论 PM可能通过激活YAP/TAZ信号通路,对PTOA大鼠具有一定的治疗作用.
Objective To investigate therapeutic effects of pristimerin(PM)on post-traumatic osteoarthritis(PTOA)rats by regulating the Yes-associated protein(YAP)/transcriptional co-activator with a PDZ-binding domain(TAZ)signaling pathway.Methods SD male rats were randomly separated into a control group,a PTOA group,a low-dose PM group(PM-L group,100 mg/kg PM),a high-dose PM group(PM-H group,200 mg/kg PM),and a high-dose PM+YAP inhibitor VTPF group(PM-H+VTPF group,200 mg/kg PM+10 mg/kg VTPF),with 12 rats in each group.Changes in the thermal pain threshold and tenderness threshold of rats were recorded.HE staining was applied to observe pathological changes in rat cartilage tissue and Mankin's score was performed.Enzyme linked immunosorbent assay(ELISA)was applied to detect inflammatory factors such as nitric oxide(NO),interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),matrix metalloproteinase-1(MMP-1),and MMP-3 in cartilage tissue.TUNEL staining was applied to observe apoptosis of cartilage tissue cells.Real time fluorescence quantitative PCR(RT-qPCR)as-say was used to detect mRNA of TAZ,IL-1β and TNF-α in rat cartilage tissue.Western Blot was applied to detect YAP/TAZ sig-naling pathway related proteins in cartilage tissue.Results Compared with control group,thermal pain threshold,tenderness threshold,YAP phosphorylation in cartilage tissue,and TAZ protein and mRNA in the PTOA group were significantly lower(P<0.05)while IL-1β,TNF-α and their mRNA,NO,MMP-1,MMP3,apoptosis rate,and Mankin's score in cartilage tissue were obviously higher(P<0.05).HE showed that the cartilage layer became thinner,the structure was unclear,and a large number of chondrocytes were lost and arranged in disorder.Compared with PTOA group,changes in relevant indicators of rats in the PM-L and PM-H groups were opposite to the above(P<0.05),and the cartilage tissue lesions were reduced.YAP inhibitor VTPF alleviated the therapeutic effect of PM on PTOA.Conclusion PM may have a certain therapeutic effect on PTOA rats by activating the YAP/TAZ signaling pathway.
余华;李少星;刘相杰
成都医学院第一附属医院,四川 成都 610500
扁蒴藤素Yes相关蛋白/转录共激活因子PDZ结合基序信号通路创伤性关节炎治疗
pristimerinYes-associated protein/transcriptional co-activator with a PDZ-binding domain signaling path-waypost-traumatic osteoarthritistreatment
《四川医学》 2024 (005)
491-496 / 6
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