目的 研究JNK信号通路在大鼠低氧高二氧化碳性肺动脉高压(HHPH)发生发展中的动态变化,探讨三七皂苷(PNS)防治HHPH的机制.方法 复制慢性HHPH的SD大鼠模型,分为正常组(N,n=10),低O2高CO2组(H4w,n=10)及PNS治疗组(Hp4w,n=10).RT-qPCR及Westem blot检测JNK mRNA及蛋白表达情况.原代培养SD大鼠肺动脉平滑肌细胞(PASMCs),取2～5代对数生长期细胞,分为正常组(N)、低O2高CO2组(H)及PNS治疗组(R10、R50、R100),PNS治疗组分别用不同浓度(10、50、100 mg/L)的三七皂苷单体R1进行处理,24 h后收集细胞,RT-qPCR及WestemBlot检测JNK mRNA及蛋白表达情况.然后用R1及JNK抑制剂SP600125分别及联合处理PASMCs 5d,CCK8检测细胞增殖隋况.结果 ①与N组相比,H4w组和Hp4w组mPAP均明显增高(P＜0.05),同时Hp4w组mPAP明显低于H4w组(P＜0.05).②肺组织JNK mRNA在H4w组中表达最高,N组最低,Hp4w组介于两组之间(均P＜ 0.05);p-JNK蛋白在H4w组较N组高表达(P＜0.05),Up4w组较H4w组下降(P＜0.05).③PASMCs H组JNK mRNA及蛋白表达明显高于N组(P＜0.05),与H组相比,R1(10、50、100 mg/L)不同程度抑制了JNK mRNA及蛋白表达(P＜0.05).④PNS及SP600125处理组PASMCs均出现了明显的增殖受抑(P＜0.05).结论 JNK信号通路参与了大鼠HHPH的形成.PNS可减轻这一过程,其机制可能与PNS抑制JNK的表达与激活有关.%Objective To investigate the dynamic change of JNK signaling pathway in the occurrence and development of hypoxic hypercapnia pulmonary hypertension in rats,discuss the mechanism of panax notoginoside (PNS) in the protection of hypoxic hypercapnia pulmonary hypertension.Methods Rats pathological models of hypoxic hypercapnia pulmonary hypertension were established:SD rats were randomly divided into three groups (n=10):normal group (N group),hypoxic hypercapnia for 4 week group (H4w group),and PNS-injected group (Hp4w group).RT-qPCR and Western blot were used to detect the expression of JNK.Primary cultured PASMCs,isolated from SD rats,were incubated in logarithmic growth phase from the 2nd to 5th generation.PASMCs were divided into five groups:normal group (N group),hypoxic hypercapnia group (H group),and R1 treated group with different concentrations (10,50 and 100 mg/L) of notoginsenoside monomer R1 under the condition of 6％ CO2 plus 1％ O2 for 24 h (R10,R50 and R100 groups).The expressions of JNK mRNA and protein levels were detected by RT-qPCR and Western blot.PASMCs were treated with R1 and JNK inhibitor SP600125 for 5 days,CCK8 was used to detect the proliferation of PASMCs.Results ①The mPAP in H4w and Hp4w group was higher than that of N group (P ＜ 0.05),but mPAP in Hp4w group was obviously lower than that of H4w group (P ＜ 0.05).②The mRNA and protein levels of JNK were significantly increased in H4w and Hp4w groups,compared with N group (P ＜ 0.05),and Hp4w group was lower than H4w group (P ＜ 0.05).③The protein and mRNA expression levels of JNK in PASMCs were significantly higher in H group than those in N group (P ＜0.05);compared with H group,in R1 treatment (10,50 and 100 mg/L) groups,the expression levels of JNK were markedly decreased (P ＜ 0.05).④The proliferation of PASMCs were significantly inhibited in groups treated with R1 and JNK inhibitor SP600125 (P ＜ 0.05).Conclusion JNK may play an important role in the development of hypoxia induced pulmonary hypertension.The effect of PNS on reducing pulmonary hypertension and improving pulmonary vascular wall remodeling may be partly related to its inhibition of JNK pathway.
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