体循环肠灌流法研究红禾麻提取物在类风湿性关节炎大鼠与正常大鼠体内的肠吸收差异

比较红禾麻提取物在AA大鼠与正常大鼠体内的肠吸收差异。实验采用体循环肠灌流法,建立大鼠RA模型,应用UPLC-MS/MS检测肠灌流液中新绿原酸、绿原酸、隐绿原酸、芦丁、异槲皮苷、槲皮苷、山奈酚-3-O-芸香糖苷、木犀草苷的含量,探究各成分在不同因素(pH值、药物浓度、肠段、胆汁和P-gp抑制剂)下的肠吸收特性。结果表明,槲皮苷的吸收方式为主动转运,其余7个成分的吸收方式为被动扩散。除绿原酸、隐绿原酸外,正常状态下各成分以回肠吸收最好,病理状态下以十二指肠吸收最好,推测药物吸收的特定部位会受疾病状态的影响而发生改变。各成分的吸收均受pH、胆汁和P-gp的影响,其中,槲皮苷可能是P-gp的底物。RA能影响红禾麻提取物在大鼠体内的肠吸收,其作用机制尚需进一步研究。

蚕砂联合光动力对类风湿性关节炎大鼠作用机制的实验研究

目的蚕砂联合光动力对类风湿性关节炎大鼠作用机制的实验研究。方法于2014年7月—2015年6月在哈尔滨市第四人民医院实验室,将40只健康SD大鼠分为正常对照组、蚕砂灌胃组、光动力组以及蚕砂联合光动力组各10只,模型对照组以及蚕砂联合光动力组给予,以建立大鼠类风湿性关节炎的动物模型,蚕砂灌胃组以及蚕砂联合光动力组予以蚕砂,正常对照组及光动力治疗组予以等剂量频次的蒸馏水。比较各组大鼠情况、踝关节左右横径测量结果、关节病理评分、滑膜液内VEGF、MMP-3及SOD水平和外周血细胞因子水平。结果发病后1~7 d蚕砂灌胃组、蚕砂联合光动力组以及蚕砂联合光动力组大鼠踝关节左右横呈持续升高趋势,发病后11~21 d 3组大鼠踝关节左右横水平降低,与正常对照组相比,差异有统计学意义(P<0.05)。与蚕砂灌胃组以及蚕砂联合光动力组比,蚕砂联合光动力组滑膜下炎症、滑膜增生、血管已形成及软骨损坏骨破坏和关节病理总分较低[(1.02±0.11)分、(1.49±0.17)分、(0.92±0.10)分、(0.10±0.01)分、(3.44±0.37)分](P<0.05)。与正常对照组相比,蚕砂灌胃组、蚕砂联合光动力组以及蚕砂联合光动力组关节滑膜液内VEGF、MMP-3水平较高(P<0.05),SOD水平较低(P<0.05);与蚕砂灌胃组以及蚕砂联合光动力组相比,蚕砂联合光动力组关节滑膜液内VEGF、MMP-3较低[(0.35±0.05)μg/L、(2.02±0.21)pmol/L](P<0.05),SOD较高(17.19±1.83)U/m L(P<0.05)。与正常对照组相比,蚕砂灌胃组、蚕砂联合光动力组以及蚕砂联合光动力组血清IL-1和TNF-α水平较高(P<0.05),TGF-β水平较低(P<0.05);与蚕砂灌胃组以及蚕砂联合光动力组相比,蚕砂联合光动力组血清IL-1和TNF-α较低[(35.39±4.17)mg/L、(23.29±2.59)mg/L](P<0.05),TGF-β较高(23.29±2.57)mg/L(P<0.05)。结论蚕砂联合光动力对类风湿性关节炎大鼠能够破坏血管翳,保护骨和软骨,缓解炎性反应,有望为临床治疗方面提供新的干预方法。

离体外翻肠囊法研究红禾麻提取物在正常与类风湿性关节炎模型大鼠中的肠吸收特性差异

考察红禾麻提取物在正常与类风湿性关节炎病理状态下大鼠体内的肠吸收特性差异,采用大鼠离体外翻肠囊模型,以红禾麻提取物5.0 g·L-1为吸收浓度,通过UPLC-MS/MS检测肠吸收液样品中新绿原酸、绿原酸、隐绿原酸、芦丁、山柰酚-3-O-芸香糖苷、木犀草苷、槲皮苷、异槲皮苷8种指标成分的含量,计算其累积吸收量(Q)和吸收速率常数(Ka),比较正常大鼠和类风湿性关节炎大鼠肠吸收液中红禾麻提取物各成分的吸收特征。结果表明,红禾麻提取物中8种指标成分均可吸收进入肠囊且成线性吸收,各成分的累积吸收量-时间曲线均为上升趋势,未呈饱和现象,其相关回归系数(R2)均大于0.92,符合零级吸收速率过程,推测各成分可能的吸收方式为被动扩散。正常状态下各成分以回肠吸收最好(除绿原酸外),病理状态下以十二指肠吸收最好,且8种成分在类风湿性关节炎大鼠各肠段的总体吸收优于正常大鼠,推测类风湿性关节炎可能会改变药物吸收的特定部位。实验表明类风湿性关节炎会改变红禾麻提取物的肠吸收,其作用机制有待进一步的深入研究。

Effect of electroacupuncture on c-IAP1 mRNA and C-IAP2 mRNA in synovial tissues of rats with adjuvant arthritis

Objective: Td observe the therapeutic effect of electroacupuncture (EA) at Zusanli (ST 36), Guanyuan (CV 4) and Ashi points on adjuvant arthritis rats, and explore the mechanism of EA treatment of rheumatoid arthritis (RA). Methods: Sixty male rats were randomly divided into a normal group, a model group, a methotrexate group and an EA group, with 15 rats in each group. Rats in the normal group and the model group were routinely raised and did not receive treatment;rats in the methotrexate group received methotrexate at a dose of 0.35 mg/(kg bw), twice a week for 3 weeks;rats in the EA group received acupuncture at Zusanli (ST 36), Guanyuan (CV 4) and Ashi points, and the bilateral Zusanli (ST 36) and Ashi points were connected to EA apparatus, once a day for 3 weeks. The general status, the swelling degree of the toe, the arthritis index (Al) score, the pathological morphology of the ankle joint, and the mRNA expressions of cellular inhibitor of apoptosis protein (c-lAP) 1 and C-IAP2 in joint synovial tissue cells of the rats in each group were observed. Results: The swelling degree of the toe, Al score and mRNA expressions of c-IAPl and C-IAP2 in the model group were significantly higher than those in the normal group (all P<0.05). Compared with the model group, the swelling degree of the toe, Al score and mRNA expressions of c-IAPl and C-IAP2 in the methotrexate group and the EA group improved (P<0.01 or P<0.05);the expressions of c-IAPl mRNA and C-IAP2 mRNA in rat synovial tissues in the EA group were significantly higher than those in the methotrexate group (P<0.01). Conclusion: EA alleviates joint swelling in rats with adjuvant arthritis. The mechanism may be related to suppress!ng mRNA expressions of c-IAPl and C-IAP2, thus to induce apoptosis of synoviocytes.

Study on the mechanism of moxibustion for rheumatoid arthritis based on liquid chromatography-mass spectrometry

Objective To explore the mechanism of moxibustion for rheumatoid arthritis(RA)by observing the metabolite changes in urine using liquid chromatography-mass spectrometry(LC-MS)-based metabolomic analysis.Methods Twenty-four rats were randomly divided into a control group,a model group,and a moxibustion group.Rats in the model and moxibustion groups were established as collagen-induced arthritis(CIA)models.The control and model groups did not receive any intervention;rats in the moxibustion group received moxibustion at Shenshu(BL23)and Zusanli(ST36).After three weeks of intervention,ankle joint,serum,and urine samples were collected for pathological examinations and metabolomic tests.Results After moxibustion treatment,the CIA rats showed increased body mass,reduced swelling of the hind paws and arthritis score,decreased serum cytokine levels,and improved histopathological evaluation of the ankle joint.Twenty-four significantly altered metabolites were found,mainly involved in alanine metabolism,taurine and hypotaurine metabolism,tricarboxylic acid cycle,phenylalanine metabolism,tyrosine metabolism,and primary bile acid biosynthesis.These metabolites may serve as potential biomarkers for RA.Conclusion Moxibustion can effectively resist inflammation in CIA rats.The potential biomarkers and the abnormal metabolic pathways of RA can be identified by LC-MS-based metabolomics.Metabolomics may be an effective way to explain the mechanism of moxibustion in treating RA.