吡格列酮对X线照射后成纤维细胞分泌的基质金属蛋白酶-9和基质金属蛋白酶抑制物-1表达的影响

目的观察并分析吡格列酮对X射线导致的小鼠肺成纤维细胞(L929细胞)中的基质金属蛋白酶-9(MMP-9)、基质金属蛋白酶抑制物-1(TIMP-1)表达的影响。方法采用RT-PCR法证实L929细胞中具有过氧化物酶体增殖因子活化受体γ(PPAR-γ)表达。采用剂量为10 Gy X射线照射L929细胞,用RT-PCR方法观察照射前与照射后0 h、2 h、7 h、24 h、48 h表达MMP-9和TIMP-1的情况。采用吡格列酮处理上述照射处理过的L929细胞,同上法观察不同时点如上两个指标的表达。结果 RT-PCR检测结果表明L929细胞中存在PPAR-γ的表达。L929细胞经10 Gy照射后其MMP-9表达随照射后时间的延长逐渐增加,在照射后48 h表达最高(F=4. 135,P=0. 022); TIMP-1随照射后时间的延长表达逐渐减少,在照射后48 h表达最低(F=3. 646,P=0. 041)。上述L929细胞经吡格列酮处理后48 h其MMP-9表达降低最明显[0. 76∶0. 52 (F=3. 735,P=0. 024)],其TIMP-1表达升高最明显[0. 17∶0. 27(F=3. 908,P=0. 032)]。结论小鼠L929细胞经X射线照射后出现纤维化相关因子MMP-9表达的升高和TIMP-1表达的降低。吡格列酮可使上述MMP-9高表达降低,使TIMP-1低表达升高。     

PPAR-γ对放射诱导成纤维细胞分泌PAI-1和TGF-β的影响

目的 分析过氧化物酶增殖体活化受体-γ(PPAR-γ)配体(匹格列酮)对X射线诱导纤溶酶原抑制物-1(PAI-1)、转化生长因子-β(TGF-β)在小鼠肺成纤维细胞(L929细胞)中的mRNA表达和L929细胞增殖活性的影响.方法 用RT-PCR证实L929细胞表达PPAR-γmRNA.分别采用10、4、2 GyX射线照射L929细胞,观察照射前后PAI-1、TGF-βmRNA的表达情况.照射+匹格列酮处理L929细胞,用RT-PCR方法观察对PAI-1、TGF-βmRNA的表达影响,用MTT法观察对细胞增殖活性影响.结果 RT-PCR检测结果显示10 Gy照射48 h后L929细胞中PAI-1、TGF-βmRNA表达明显升高[0.18∶0.78、0.22∶0.76(F=3.70,P=0.010)],2 Gy和4 Gy有类似趋势[0.18∶0.43、0.18∶0.44和0.22∶0.39、0.22∶0.40(F=3.40,P=0.090)];10 Gy照射+匹格列酮48 h后明显降低[0.78∶0.45、0.76∶0.54(F=3.90,P=0.010)],2 Gy和4 Gy的也有类似趋势[0.43∶0.37、0.44∶0.35和0.39∶0.32、0.40∶0.32(F=2.40,P=0.210)].MTT结果显示与空白对照比较2、4、10 Gy照射和单用匹格列酮均会抑制L929细胞增殖[0.44∶0.39、0.36、0.34和0.32(F=3.90,P=0.040)],2、4、10 Gy照射+匹格列酮更能抑制L929细胞增殖[0.44∶0.27、0.26、0.25(F=2.50,P=0.005)].结论 X射线照射L929细胞后出现促纤维化因子PM-1、TGF-β表达升高.匹格列酮使PAI-1、TGF-β高表达降低,也会抑制成纤维细胞的增殖.

Abstract：

Objective To observe the influence of peroxisome proliferator activated receptor-γligand (PPAR-γ, pioglatazone) on expression of PAI-1 and TGF-β mRNA and proliferation in fibroblast cells before and after X-ray radiation, and to study the effect of PPAR-γon normal cells during radiation induced fibrosis process. Methods RT-PCR method was used to measure PPAR-γgene expression in L929 cells.After X-ray irradiation of 10 Gy,4 Gy or 2 Gy, the expressions of PAI-1 and TGF-β mRNA in mouse lung fibroblast cells (L929) were measured using RT-PCR. After X-ray irradiation and pioglatazone treatment,the influence of pioglatazone on PAI-1 and TGF-β was measured using RT-PCR method. MTT method was used to test cell proliferation after the treatment of irradiation and pioglatazone. Results PPAR-γ mRNA expression was observed in L929 cells. Expression of PAI-1 and TGF-β mRNA reached the highest level 483.40,P =0. 090) ). At 48 h after the treatment of pioglatazone and 10 Gy radiation, pioglatazone decreased 0. 36, 0. 34 and 0. 32( F = 3.90, P = 0. 040) ). The inhibitory effect was significantly increased when L9292. 50,P =0. 005)). Conclusions X-ray irradiation can increase the expression of PAI-1 and TGF-β in L929 cells. Pioglatazone can decrease the expression of radiation-induced PAI-1 and TGF-β, and restrain the fibroblast proliferation.     

头颈部骨肉瘤治疗现状

头颈部骨肉瘤为少见肿瘤,临床特点与复发模式不同于其他部位骨肉瘤。头颈部骨肉瘤发病年龄晚,局部复发率高,远处转移率低,局部复发为死亡主要原因。手术为主要治疗手段。手术切缘阳性、近切缘及手术切缘不确定者推荐术后放疗。化疗的作用尚有争议。针对复发转移、不可手术切除的骨肉瘤,有效的分子靶向治疗药物有待于继续探索。     

吡格列酮调节AMPK/mTOR信号通路对肺癌A549细胞顺铂耐药的影响

目的 探究吡格列酮(PIO)调节腺苷酸活化蛋白激酶(AMPK)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路对肺癌A549细胞顺铂(CDDP)耐药性的影响。方法 将肺癌CDDP耐药细胞A549/CDDP随机分为对照组(Control组)、PIO组(10μmol/L PIO)、CDDP组(10μg/mL CDDP)、CDDP+低浓度PIO组(CDDP+L-PIO组,10μg/mL CDDP+5μmol/L PIO)、CDDP+高浓度PIO组(CDDP+H-PIO组,10μg/mL CDDP+10μmol/L PIO)和CDDP+高浓度PIO组+AMPK激活剂AICAR组(CDDP+H-PIO+AICAR组,10μg/mL CDDP+10μmol/L PIO+20 mmol/L AICAR)。CCK-8法检测细胞增殖能力;划痕实验检测细胞迁移能力;Transwell实验检测细胞侵袭能力;流式细胞术测定细胞凋亡率;Western Blot检测各组细胞AMPK/mTOR通路蛋白和凋亡相关蛋白Bcl-2、Bax、Caspase-3蛋白表达。结果 与Control组相比,PIO组A549/CDDP细...     

Influence of PPAR -γligand on radiation - induced PAI - 1 and TGF -βmRNA in fibroblast cells

Objective To observe the influence of peroxisome proliferator activated receptor-γligand (PPAR-γ, pioglatazone) on expression of PAI-1 and TGF-β mRNA and proliferation in fibroblast cells before and after X-ray radiation, and to study the effect of PPAR-γon normal cells during radiation induced fibrosis process. Methods RT-PCR method was used to measure PPAR-γgene expression in L929 cells.After X-ray irradiation of 10 Gy,4 Gy or 2 Gy, the expressions of PAI-1 and TGF-β mRNA in mouse lung fibroblast cells (L929) were measured using RT-PCR. After X-ray irradiation and pioglatazone treatment,the influence of pioglatazone on PAI-1 and TGF-β was measured using RT-PCR method. MTT method was used to test cell proliferation after the treatment of irradiation and pioglatazone. Results PPAR-γ mRNA expression was observed in L929 cells. Expression of PAI-1 and TGF-β mRNA reached the highest level 483.40,P =0. 090) ). At 48 h after the treatment of pioglatazone and 10 Gy radiation, pioglatazone decreased 0. 36, 0. 34 and 0. 32( F = 3.90, P = 0. 040) ). The inhibitory effect was significantly increased when L9292. 50,P =0. 005)). Conclusions X-ray irradiation can increase the expression of PAI-1 and TGF-β in L929 cells. Pioglatazone can decrease the expression of radiation-induced PAI-1 and TGF-β, and restrain the fibroblast proliferation.