汉防己甲素片联合吡非尼酮治疗肺间质纤维化的临床研究

目的研究汉防己甲素片联合吡非尼酮胶囊治疗肺间质纤维化的疗效。方法选取2017年8月-2019年8月来郑州大学附属郑州中心医院治疗的80例肺间质纤维化患者为研究对象,所有患者随机分为对照组和治疗组,每组各40例。对照组患者口服吡非尼酮胶囊,200 mg/次,在两周时间内每次增加200 mg最后达到每次维持剂量为600 mg/次,3次/d。治疗组在对照组基础上口服汉防己甲素片,2片/次,3次/d。两组患者持续治疗2个月。观察两组的临床疗效,比较两组临床症状缓解时间、肺功能指标、血清转化生长因子-β(TGF-β1)、血管内皮生长因子(VEGF)、胰岛素生长因子-I(IGF-I)、Ⅲ型胶原(Ⅲ-C)、Ⅳ型胶原(Ⅳ-C)、透明质酸(HA)水平。结果治疗后,治疗组总有效率95.00%,显著高于对照组的77.50%(P<0.05)。治疗后,治疗组胸憋、喘息、气短、咳嗽、咳痰等症状缓解时间明显短于对照组(P<0.05)。治疗后,两组患者最大呼气流量(PEF)、用力肺活量(FVC)、一秒用力呼气容积(FEV1)水平显著升高(P<0.05),且治疗组患者PEF、FVC、FEV1水平升高较多(P<0.05)。治疗后,两组TGF-β1、VEGF、IGF-I水平显著降低(P<0.05);并且治疗组TGF-β1、VEGF、IGF-I水平降低较多(P<0.05)。治疗后,两组Ⅲ-C、Ⅳ-C、HA水平显著降低(P<0.05);并且治疗组Ⅲ-C、Ⅳ-C、HA水平降低较多(P<0.05)。结论汉防己甲素片联合吡非尼酮胶囊治疗肺间质纤维化具有较好的治疗效果,能够改善患者肺功能,缩短临床症状缓解时间,调节血清炎性因子水平,安全性较高,值得在临床上推广应用。     

Predictors of the clinical effects of pirfenidone on idiopathic pulmonary fibrosis

BackgroundIdiopathic pulmonary fibrosis (IPF) is a progressive interstitial lung disease with a poor prognosis. Recently, pirfenidone was reported to slow the rate of decline in vital capacity and improve progression-free survival in IPF. The purpose of this study was to clarify the factors that predicted a good response to pirfenidone, as well as its adverse effects.MethodsForty-one IPF cases, treated with pirfenidone from January 2009 to January 2011, were enrolled in this investigation. Disease severity was classified into grades I–IV, as defined by the Japanese Respiratory Society (JRS). Short-term responsiveness to pirfenidone was evaluated by the modified criteria of the JRS. Predictors of nausea, anorexia, or both that represented important adverse effects were examined by multivariate Cox proportional hazard analyses. Predictors of short-time responsiveness were examined by multivariate logistic regression analyses.ResultsDiagnosed by a surgical lung biopsy (SLB), the mild cases of grade I/II were predictors of good, short-term responsiveness. Patients taking acid-secretion inhibitors, including proton pump inhibitors and histamine H2-receptor antagonists, showed less anorexia, nausea, or both. Only 1 case was administered drugs to activate gastrointestinal motility.ConclusionsWe concluded that IPF patients with a mild disease, diagnosis by SLB, or both showed indications of a good response to pirfenidone. In addition, acid-secretion inhibitors may reduce the frequency of anorexia, nausea, or both from pirfenidone.     

吡非尼酮对内皮细胞间质转化的抑制作用及机制

目的:建立缺氧诱导内皮细胞的内皮-间质转化(EndoMT)模型,探讨吡非尼酮(PFD)在视网膜下纤维瘢痕形成过程中的抗纤维化作用及机制。方法:原代培养人脐静脉内皮细胞,鉴定后取4~7代用于实验。CoCl2诱导细胞缺氧建立纤维化模型。CCK-8法检测细胞增殖率,筛选药物浓度。将细胞分为对照组(无血清培养基培养)、CoCl2(200μmol/L)模型组、CoCl2+低浓度(0.3mg/mL)PFD组、CoCl2+高浓度(0.6mg/mL)PFD组。Western blot法检测细胞CD31、VE-cadherin、α-SMA、FSP1、p-p38和p38的蛋白表达水平。CD31/α-SMA免疫荧光双染法观察蛋白表达的变化。划痕实验观察细胞迁移能力的改变。q-PCR法检测TGF-β1、SNAI1 mRNA转录水平。结果:与CoCl2模型组相比,PFD能明显提高缺氧细胞的增殖率,抑制细胞的迁移能力(均P<0.05);PFD组细胞标志蛋白CD31、VE-cadherin表达增加,α-SMA、FSP1表达降低(均P<0.05)。免疫荧光检测显示PFD可明显抑制α-SMA和增加CD31的蛋白表达(P<0.05)。内皮细胞EndoMT过程中,p38总蛋白表达不变(P>0.05),但p-p38磷酸化蛋白表达增加、TGF-β1和SNAI mRNA转录水平增高的现象可明显被PFD抑制(均P<0.05)。高低浓度PFD组上述各现象无明显差异。结论:PFD可以抑制内皮细胞纤维化的发生,TGF-β/p38 MAPK通路可能是PFD调控EndoMT过程的机制之一,为视网膜下纤维化的治疗提供了新思路。     

吡非尼酮抑制大鼠角膜急性碱烧伤后新生血管的实验研究

目的研究吡非尼酮(PFD)对大鼠角膜急性碱烧伤后角膜新生血管(CNV)的抑制作用。 方法碱烧伤法制备CNV模型,模型制作成功后,将健康的斯泼累格·多雷(SD)大鼠20只随机平均分为PFD组和磷酸缓冲盐溶液(PBS)组,PFD组给予3 mg/ml PFD滴眼液,PBS组给予磷酸缓冲盐溶液,4次/d、连续14 d。裂隙灯显微镜观察角膜炎症反应评分、角膜混浊程度和角膜新生血管生长情况。第14 d处死大鼠取角膜,行苏木精－伊红(HE)染色法染色观察角膜病理形态,免疫组化CD34染色检测角膜微血管密度(MVD)。角膜炎症反应和新生血管情况在不同时间点,PFD和PBS两组间的比较采用两因素重复测量方差分析。 结果第14 d PFD组的角膜混浊评分是(1.00±0.00)分,PBS组的角膜混浊评分是(3.40±0.52)分,PFD组的角膜混浊评分低于PBS组,差异有统计学意义(t＝－14.697,P<0.05)。第7 d和第14 d PFD组的角膜炎症评分分别是(3.50±0.53)分和(2.2±0.42)分,PBS组的角膜炎症评分分别是(4.9±0.57)分和(3.3±0.48)分,PFD组的角膜炎症评分均低于PBS组,差异有统计学意义(t＝－5.715,－5.425;P<0.05)。PFD组,随时间延长,角膜透明度增加,炎症下降。该组第7 d与第3 d比较,差异有统计学意义(t＝7.56,P<0.05)。第14 d分别与第3 d及第7 d比较,差异均有统计学意义(t＝13.17,6.08;P<0.05)。PBS组,第7 d与第3 d比较,差异有统计学意义(t＝2.64,P<0.05)。第14 d分别与第3 d及第7 d比较,差异均有统计学意义(t＝8.74,6.79;P<0.05)。第3 d、第7 d及第14 d PFD组的CNV面积分别是(6.06±0.93)mm²、(17.94±1.89)mm²及(23.89±1.84)mm²,PBS组的CNV面积分别是(9.08±1.42)mm²、(27.11±2.02)mm²及(31.01±2.04)mm²,PFD组的CNV面积均低于PBS组,差异均有统计学意义(t＝－5.609,－10.452,－8.202;P<0.05)。PFD组大鼠角膜CNV模型的CNV面积,第7 d比第3 d增大,差异有统计学意义(t＝17.83,P<0.05);第14 d比第3 d及第7 d,差异均有统计学意义(t＝27.38,7.13;P<0.05)。PBS组,第7 d与第3 d的CNV面积比较,差异有统计学意义(t＝23.09,P<0.05);第14 d比第3 d及第7 d,差异均有统计学意义(t＝27.90,4.29;P<0.05)。HE病理染色显示PBS组的角膜基质有大量炎症细胞浸润及纤维组织增生,而PFD组炎症细胞数量极少,基质排列较规则。免疫组化CD34染色结果显示PFD组的MVD为(3.17±1.17)条/mm²,PBS组的MVD为(10.83±2.48)条/mm²,PFD组的MVD低于PBS组,差异有统计学意义(t＝－6.842,P<0.05)。 结论PFD能减轻大鼠角膜急性碱烧伤模型中的角膜炎症反应,并抑制角膜新生血管形成,有望为临床防治角膜新生血管类疾病提供一种新方法。     

Management of idiopathic pulmonary fibrosis

Idiopathic pulmonary fibrosis (IPF) is a deadly progressive lung disease without an effective standard treatment approach. Because of the complexity and uncertainties of IPF treatment, therapeutic decisions need to be tailored to the individual patient, after discussing the potential benefits and pitfalls. Pirfenidone has been approved for the treatment of IPF in many countries, but is not recommended as a first-choice therapy by current guidelines because of the lack of a definite efficacy. Randomized controlled trials represent a valid choice for patients with IPF, and their completion is important in improving both survival and quality of life.     

Protective effects of pirfenidone on D-galactosamine and lipopolysaccharide-induced acute hepatotoxicity in rats

OBJECTIVE AND DESIGN: To investigate the protective effects of pirfenidone on acute liver damage caused by D-galactosamine (GalN)/lipopolysaccharide (LPS) in rats. MATERIAL AND TREATMENT: Sprague-Dawley rats were divided into five groups (five rats per group): normal control group, GalN/LPS-treated group, and three pirfenidone-treated group (100, 300 and 500 mg/kg i.p., respectively). All biochemical and histological indexes were determined at 12 h after GalN/LPS challenge. METHODS: Severity of liver injury was assessed by determination of serum ALT, AST levels and histological analysis. SOD activity and MDA concentrations as well as TNF-alpha and IFN-gamma levels in the liver of rats were measured. The expression of iNOS and its product, NO concentration were also determined. RESULTS: Pretreatment with pirfenidone significantly attenuated GalN/LPS-induced severe hepatotoxicity, as evidenced by decreased ALT, AST levels and MDA content and improved histopathological changes. Pirfenidone inhibited the elevated levels of TNF-alpha and IFN-gamma and reduced the induction of iNOS/NO in a dose-dependent manner, which might be important mechanisms related to its protective effect. CONCLUSIONS: Pirfenidone can provide a definite protective effect against acute hepatic injury caused by GalN/LPS in rats, which may be mainly mediated through its anti-inflammatory effect.