Objective To investigate the role of the PGE2 and EP2/EP3 receptor on improving lung function of rats with pulmonary fibrosis treated with Youguiyin Decoction. Methods 32 SPF SD male rats were divided into normal group, model group, Youguiyin low dose group and Youguiyin high dose group, according to the random number table method. The normal group (8 rats) was given no intervention and was given normal feeding. The other 3 groups were established by intravenous injection of Bleomycin (6 mg/kg) via tracheal intubation. Youguiyin low dose group (8 rats) and Youguiyin high dose group (8 rats) were given 5 g/kg and 10 g/kg of Youguiyin Decoction at specific time from the 14 th day after model establishment. The model group (8 rats) was given the same dose of drinking water once a day for 20 days. The lung weight and body weight were recorded, and the lung coefficient was calculated. The concentration of PGE2 in serum and lung tissue was measured with ELISA. The expression of EP2 and EP3 in lung tissue and tissue localization were analyzed by immunohistochemical staining, and the number of positive cells was counted. Results Compared with the normal group, the lung weight (3. 1 ± 0. 44 g, P ＜ 0. 05) and lung coefficient (0. 69 ± 0. 13％, P ＜0. 05) of the model group increased pathologically, and the level of PGE2 increased in both serum (45. 49 ± 7. 38 pg/ml, P ＜ 0. 05) and lung tissue (113. 64 ± 16. 78 pg/ml, P ＜ 0. 05). Abnormal proliferation of AEC IIs associated with strong EP2-positive in the consolidation of lung tissues, and the macrophages accumulated in the alveolar ducts and alveolar spaces were EP3-positive. The positive cells in the two groups increased significantly (P ＜ 0. 05). Compared with the model group, the lung weight (2. 63 ± 0. 24 g, P ＜ 0. 05) and lung coefficient (0. 57 ± 0. 08％, P＜ 0. 05) of Youguiyin low dose group decreased, and the level of PGE2 decreased in both serum (23. 01 ± 8. 61 pg/ml, P ＜ 0. 05) and lung tissue (89. 84 ± 20. 54 pg/ml, P ＜ 0. 05). The expression of EP2 in the AEC IIs with strong EP2-positive decreased, and the number of cell reduced (P ＜ 0. 05). The number of macrophages with EP3-positive increased (P ＜ 0. 05). In the Youguiyin high dose group, there was no statistical difference except the decreased level of PGE2 in serum (P ＜ 0. 05). Conclusion Youguiyin can significantly improve pulmonary fibrosis induced by Bleomycin in rats, which may be associated with lowing PGE2 level in the consolidation of lung tissues, inhibiting the expression of EP2 in the consolidation area, reduction of AECIIs with EP2-positive, and increase of macrophages with EP3-positive.

Objective To investigate the relative mechanism of Maimendong Tang (Ophiopogon Decoction) in amelioration of lung function and endoplasmic reticulum stress (ERS) in rats with bleomycin-induced pulmonary fibrosis.Methods Male healthy SD rats were randomly divided into normal group, model group, Maimendong Tang group 1 and Maimendong Tang group 2.The rat model of pulmonary fibrosis was established by bleomycin pulverization via tracheal intubation.On the 14 t h d after modeling, Maimendong Tang was intragastrically given to Maimendong Tang group1 and Maimendong Tang group 2 twice a day for 20 d continuously.The general conditions were observed, and pathological changes of lung tissue were observed after HE and Masson staining.The forced vital capacity (FVC) , 0.4 second rate (FEV0.4/FVC) , mass FVC, inspiratory resistance (Ri) , expiratory resistance (Re) and dynamic pulmonary compliance (Cydn) were detected by using spirometer.The expressions of glucose regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP) protein in lung tissue were analyzed by using Western blotting assay and immunohistochemistry staining technique.Results Compared with normal group, FVC, FVC/lung mass and Cydn decreased, and 0.4 second rate increased in model group.Compared with model group, FVC increased and 0.4 second rate decreased and Cydn increased in Maimendong Tang group 1 and Maimendong Tang group 2, and mass FVC increased in Maimendong Tang group 2.Pathological sections showed that there were chronic inflammatory cell infiltration in consolidation zone of lung tissue and massive collagen deposition, GRP78 and CHOP with high expressions were mainly positioned in vesicular type Ⅱ alveolar epithelial cells (AECⅡs).The inflammation was relieved, collagen deposition was decreased, protein expressions of GRP78 and CHOP were reduced and in Maimendong Tang group 2.Conclusion Maimendong Tang can significantly improve lung function and reduce lung interstitial collagen deposition, which may be related to regulation of protein expressions of GRP78 and CHOP in AECⅡs in consolidation zone of lung tissue, relief of ERS, and recovery of normal AECⅡs function.

目的 以体外培养的肥大细胞为实验材料,考察注射用双黄连诱发肥大细胞脱颗粒早期的生物学效应,初步探讨中药注射剂安全性评价的质控指标.方法 肥大细胞为RBL-2H3细胞株(ATCC-6562),注射用双黄连购自哈药集团中药二厂(Z20043425);细胞活力检测采用MTT法,β-氨基己糖苷酶检测采用ELISA法;采用扫描电镜观察肥大细胞形态,用激光共聚焦显微镜检测活细胞内钙离子浓度.结果 注射用双黄连细胞毒性剂量及致类过敏剂量为≥0.08 mg/mL;肥大细胞脱颗粒反应出现在药物作用早期(≤1分钟);较低浓度注射用双黄连诱发肥大细胞内钙离子浓度峰值出现在4分钟左右.结论 注射用双黄连作用肥大细胞早期,可激活Ca2+相关信号通路、启动肥大细胞脱颗粒反应、大量释放β-氨基己糖苷酶.该结果提示:基于RBL-2H3细胞的Ca2+浓度及致敏介质检测平台有望成为中药注射剂安全性评价的质控指标之一,具有重要应用前景.

目的:探讨温补肾阳的代表方剂右归饮对博莱霉素致肺纤维化大鼠的药效作用和相关机制.方法:SPF级雄性SD大鼠25只,采用随机数字法,将SD大鼠分为正常组、模型组、右归饮高、中、低剂量组,每组各5只.气管内注射博莱霉素(5 mg·kg-1)建立实验性肺纤维化模型,并于造模后第15天开始灌胃高、中、低剂量(20,10,5 g·kg-1)右归饮,连续给14 d.用动物肺功能仪检测大鼠吸气阻力(Ri),呼气阻力(Re),肺顺应性(Cdyn),用力肺活量(FVC),0.4s率(FEV04/FVC％),呼气峰值流速(PEF),最大呼气中期流速(MMF),计算质量FVC;称量脏器质量和大鼠体质量,计算脏器系数;用血气分析仪检测腹主动脉酸碱度(pH),氧分压(PO2),二氧化碳分压(PCO2),氧饱和度(SO2);用酶联免疫吸附法(ELISA)检测血清中肾素,前列腺素E2(PGE2),基质金属蛋白酶-1(MMP-1)含量;取左肺进行马松(Masson)染色观察肺组织的病理变化.结果:与正常组比较,气管内注射5 mg·kg-1博莱霉素能够明显增加SD大鼠肺脏质量和肺脏系数(P＜0.05),降低Cydn,FVC和质量FVC(P ＜0.05),增加Ri(P＜0.05),升高FEV04/FVC％(P＜0.05),升高腹主动脉血中PCO2水平(P＜0.05),明显降低PO2,SO2水平(P＜0.05),升高血清中PGE2水平(P＜0.05),病理显示模型大鼠肺组织形成大量实变区,发生实变的肺组织以细胞异常增殖为主的非纤维化区和伴有大量细胞外基质沉积的纤维化区为主.与模型组相比,右归饮中剂量组肺脏质量明显回落(P＜0.05),Cydn,FVC和质量FVC回升(P＜0.05),Ri,FEV0.4/FVC％下降(P＜0.05),血清中PGE2水平下降(P＜0.05);右归饮高剂量组能够明显升高动脉血中PO2,SO2水平(P＜0.05).病理显示右归饮中剂量组能够抑制肺组织实变区向非实变区的炎性浸润.结论:右归饮能够改善实验性肺纤维化SD大鼠的肺功能,与右归饮降低血清中PGE2水平,抑制炎性浸润区的扩散,延缓纤维化进程有关.

目的 探讨姜黄素对转化生长因子β1(TGF-β1)诱导的大鼠肾间质成纤维细胞(NRK49F)中小分子RNA-21(miR-21)及纤维化相关因子表达的影响.方法 将NRK49F细胞分为正常组、模型组、姜黄素组.模型组以TGF-β1(10μg/L)诱导NRK49F细胞活化,姜黄素组以高、中、低浓度(20、10、5μmol/L)干预24 h和72 h.免疫荧光方法及高内涵成像分析设备检测并分析NRK49F细胞中α-平滑肌肌动蛋白(α-SMA)表达的变化;RT-PCR方法检测NRK49F细胞中miR-21及纤维化相关因子COL1 A1、COL3 A1、α-SMA、Smad3 mRNA表达的变化.结果 TGF-β1作用于NRK49 F细胞72 h后,能明显上调细胞中α-SMA蛋白的表达(P<0.01);与模型组相比,姜黄素各组的α-SMA蛋白表达明显下调(P<0.01、P<0.01、P<0.05).TGF-β1作用于NRK49F细胞24 h后,能明显上调细胞中miR-21、COL1A1、COL3A1、α-SMA、Smad3 mRNA的表达(P<0.01);与模型组相比,姜黄素高、中浓度组可明显抑制miR-21、COL1 A1、COL3 A1、α-SMA、Smad3 mRNA的表达(P<0.01或P<0.05);姜黄素低浓度组可明显抑制COL1A1、COL3A1、α-SMA mRNA的表达(P<0.01),对miR-21、smad3 mRNA的表达有下调的趋势.结论 姜黄素能够有效抑制TGF-β1诱导的NRK49F细胞中纤维化相关因子的基因及蛋白表达,其作用与下调miR-21的表达相关.

Objective:To observe different effects of Maimendong Decotion and Gancao GanjiangDecotionin on pulmonary fibrosis rat in the stage of fibrotic phase.Methods:After 14 days of establishing rat model by intratracheal injection of bleomycin,Maimendong decotion(7.5 g/kg,15 g/kg,30 g/kg) and Gancao Ganjiang decotion(3.75 g/kg,7.5 g/kg,15 g/kg) were given by gavage once a day for 2 week.Meanwhile,the model group and the normal group were administered with distilled water of same quantity.After gavage of decotion for 14 days,the forced vital capacity (FVC),the indexes of lung dynamic compliance(Cdyn),the ratio of FEV 0.4 to forced vital capacity(FEV0.4/FVC%),peak expiratory flow rate (PEF),maximal mid-expiratory flow rate (MMF) and the blood gas changes were recorded,then the organ coefficients were calculated,lung tissue and airway pathologic changes were observed by HE staining.Results:Compared with the model group,the high dose group of Maimendong decotion reduced the lung weight and lung index significantly(P<0.05),and increased Cdyn,FVC and FVC/lung weight significantly(P<0.05).The Cdyn and FVC of high dose group of Gancao Ganjiang decotion were higher than the model group(P<0.05).The intermediate-dose Gancao Ganjiang decotion improved FVC/lung weight (P<0.05) and reduced the rate of FEV0.4/FVC% significantly (P<0.05) compared with the model group.Gancao Ganjiang decoction improved PO2 and SO2 significantly compared to model group(P<0.05).Conclusion:Both Maimendong decotion and Gancao Ganjiang decotion are better for the rat model of bleomycin induced pulmonary fibrosis.The former can reduce the interstitial hyperplasia around the airway in fibrosis area,and the latter protect the structure and function of the alveolar in non-fibrosis area.

Objective To observe the effect of spleen deficiency syndrome induced by rhubard solution combined with lung fibrosis induced by bleomycin on lung function of rats, so as to discuss the relationship between spleen deficiency syndrome (SDS) and pulmonary fibrosis.Methods SD rats were randomly divided into normal group, SDS group, model group, SDS + lung fibrosis group.The rat SDS model was induced by intragastrical administration of rheum officinale with fasting every other day for 30 days.Then the rats were infused bleomycin(5 mg/kg)through tracheal intubation to simulate the lung fibrosis disease with SDS.Four weeks after injection of bleomycin, rats' lung function was detected, lung index and spleen index were calculated, and the pathological changes of lung tissue and aiway were observed by using HE staining and Masson staining.Results Compared with the normal group, weight of lung, lung index, FEV0.4/FVC% were increased in the model group(P<0.05), and the lung dynamic compliance (Cdyn), forced vital capacity (FVC), FVC /lung weight decreased, peak expiratory flow rate (PEF) as well as maximal mid-expiratory flow rate (MMF) decreased in model group(P<0.05).What bleomycin caused was characterized by airway centered interstitial fibrosis.In the small airway, the shed epithelial cells and mucus plugs could been seen.The lung function of rats of SDS group had no difference with that of the healthy cats.Compared with the model group, FEV0.4/FVC% significantly increased (P<0.05) and the area ratio of interstitial fibrosis increased in the SDS+ lung fibrosis group.Conclusion Spleen deficiency syndrome is a potential adverse factor to aggravate restriction of ventilation function due to bleomycin.