Suppression of puerarin on polymethylmethacrylate-induced lesion of peri-implant by inhibiting NF-κB activation in vitro and in vivo

Puerarin (PR), a natural isoflavone isolated from Chinese traditional plant pueraria lobata, has attracted considerable attention due to its important biological and pharmacological activities. However, its effects on lesion of peri-implant and related mechanism of action are still not clear, which require further investigation. In this study, we evaluated the effects of PR on polymethylmethacrylate (PMMA)-induced lesion of peri-implant in vitro and in vivo, and explored its possible mechanism of action. Our results indicated that PR could inhibit PMMA-induced osteoclastogenesis in RAW264.7 cells with a dose-dependent manner in vitro and effectively down-regulate mRNA and protein expressions of matrix metalloprotein 9 (MMP-9), tumor necrosis factor (TNF)-α, interleukin (IL)-6, and receptor activator of nuclear factor (NF)-κB (RANK), primarily via the suppression of NF-κB signaling. Furthermore, we found that PMMA induction could directly cause the phosphorylation of IκB and significantly promote the nuclear translocation of p65 in RAW264.7 cells. In other words, PR was able to dose-dependently attenuate the PMMA-induced nuclear translocation of p65 in RAW264.7 cells. In vivo, PR was observed to attenuate PMMA-induced osteoclastogenesis, osteolysis, mRNA expressions of receptor activator of nuclear factor (NF)-κB ligand (RANKL) and RANK, as well as protein levels of MMP-9, TNF-α, IL-6, and p65 in a murine calvarial osteolysis model. These findings suggested that PR might be a potential therapeutic drug to lesion of peri-implant, and provided new insights for understanding its possible mechanism.     

高效液相-示差折光法同时测定葛根素葡萄糖注射液中葛根素和葡萄糖的含量

目的： 建立同时测定葛根素葡萄糖注射液中葛根素和葡萄糖含量的高效液相-示差折光法。 方法： 采用Agilent ZORBAX NH2色谱柱(4.6 mm×250 mm,5μm),流动相为乙腈-水(80:20),柱温为30℃,流速为1.2mL.min-1,进样量为20μL,检测器为RID,检测池温度为40℃。 结果： 在选定的色谱条件下葛根素在0.1311～1.049mg.mL-1的浓度范围内与其峰面积呈线性关系(r=0.9996),葡萄糖在3.126～25.01mg.mL-1 的浓度范围内与其峰面积呈线性关系(r=0.9998),葛根素的平均回收率为100.2%,其RSD=1.0%(n=9),葡萄糖的平均回收率为100.3%,其RSD=1.1%(n=9)。结论: 本方法简便,快速,准确,重现性好,适用于葛根素葡萄糖注射液中葛根素和葡萄糖的质量控制     

葛根素对臂丛神经根性撕脱伤脊髓前角iNOS、CGRP蛋白表达及PI3K/Akt信号通路的影响

目的　探讨葛根素对臂丛神经根性撕脱伤（brachial plexus root avulsion injury,BPRAI）脊髓前角iNOS、CGRP蛋白表达及PI3K/Akt信号通路的影响。 方法　将50只雄性SD大鼠随机分为正常组、模型组、葛根素低、中、高剂量治疗组,每组10只。模型组,葛根素低、中、高剂量治疗组进行BPRAI造模,撕脱大鼠右侧C5~7脊神经前根,后根剪断,术后3个治疗组予腹腔注射葛根素,剂量分别为50、100、200 mg·kg-1·d-1,正常组、模型组腹腔注射等体积生理盐水,持续4周。采用尼氏染色、免疫荧光化学、Western blot方法,观察损伤侧脊髓前角α运动神经元（alpha motorneurons,α-MNs）的存活率,iNOS、CGRP、PI3K/Akt通路相关蛋白的表达。 结果　第4周时,低、中、高剂量的葛根素治疗可抑制α-MNs丢失（P<0.05或P<0.01）;中、高剂量的葛根素治疗可抑制iNOS表达（P<0.05）;高剂量的葛根素治疗可促进CGRP蛋白表达（P<0.05或P<0.01）;低、中、高剂量葛根素均可显著抑制p-Akt1/2/3表达（P<0.01）。 结论　葛根素可改善BPRAI造模引起的α-MNs死亡,其机制可能与葛根素能抑制iNOS蛋白的表达、促进CGRP蛋白的表达有关,并且PI3K/Akt信号通路参与其调控。     

葛根素注射液联合多奈哌齐治疗帕金森病的疗效观察

目的探讨葛根素注射液联合盐酸多奈哌齐片治疗帕金森病的临床疗效。方法选取2016年2月—2017年12月南阳市中心医院收治的帕金森病患者98例为研究对象,所有患者随机分为对照组和治疗组,每组各49例。对照组患者睡前口服盐酸多奈哌齐片,1片/次,1次/d,维持1个月,然后根据治疗效果增加剂量至2片/次,1片/d。治疗组在对照组基础上静脉滴注葛根素注射液,400 mg加入到5%葡萄糖溶液500 m L中,1次/d。15 d为1个疗程,两组患者连续治疗3个疗程。观察两组的临床疗效,比较两组的改良Webster症状评分。结果治疗后,对照组和治疗组的总有效率分别为79.59%、91.83%,两组比较差异有统计学意义(P0.05)。治疗后,两组患者改良Webster症状评分均显著下降,同组治疗前后比较差异具有统计学意义(P0.05);且治疗组患者改良Webster症状评分显著低于对照组,两组比较差异具有统计学意义(P0.05)。结论葛根素注射液联合盐酸多奈哌齐片治疗帕金森病具有较好的临床疗效,可改善临床症状,安全性较好,具有一定的临床推广应用价值。     

葛根素注射剂杂质研究

目的 研究葛根素注射剂的杂质谱,并对杂质进行结构鉴定。方法 建立高效液相梯度洗脱法,分析11个厂家共12批次葛根素注射剂杂质谱,利用半制备高效液相对注射剂中的杂质进行分离并结合核磁技术进行杂质结构确认。结果 不同生产厂家及批次的葛根素注射剂杂质种类及含量差别明显,共分离并鉴定了注射剂中的5个杂质,分别为3''-羟基葛根素、3''-甲氧基葛根素、染料木素-8-葡萄糖苷、新葛根素B、新葛根素A,均为葛根素异构体或其衍生物。结论 采用此方法检出的杂质峰数目较现行方法多,且精密度高、重复性好。通过对葛根素注射剂杂质谱的研究,可以有目的地控制部分杂质,为质量控制研究提供参考。     

近红外光谱技术结合竞争自适应重加权采样算法用于中药定量分析

目的:基质复杂、谱带重叠严重,影响了中药近红外定量模型的准确性。为解决以上问题,探讨竞争自适应重加权采样(Competitive adaptive reweighted sampling,CARS)变量筛选方法在中药材、中药提取物和中成药的定量分析中的应用。方法:采集葛根药材、葛根提取物和愈风宁心滴丸的近红外漫反射光谱,测定葛根素含量。分别优化光谱前处理方式,剔除奇异样本后,运用CARS法筛选出的相关变量,建立偏最小二乘法(PLS)校正模型。结果:原料、中间体和制剂的定量模型交互验证均方差(RMSECV)分别为0.35%,1.76%,0.54%,与基于全光谱建立的模型比较,原料、中间体和制剂的CARS-PLS模型的预测准确度均有提高。结论:竞争自适应重加权采样变量筛选方法可以提高模型的预测能力,并有效简化运算过程,为中药的快速、无损检测提供了新的思路。     

葛根素对心房纤颤患者血小板活化和内皮功能的影响

目的 :观察葛根素对心房纤颤患者血小板活化和内皮功能的影响。方法 :选择 42例心房纤颤患者 ,随机分为葛根素组和阿斯匹林组两组。另选 2 1例体检健康者作为正常对照组。在常规治疗基础上 ,葛根素组用 5 %葡萄糖注射液 葛根素 5 0 0mg静脉滴注 ,1次 /d ,连续 2周 ;阿斯匹林组用阿斯匹林 15 0mg ,1次 /d ,连续口服 2周。观察治疗前后血小板活性标志物P选择素和内皮功能受损标志物血浆血管性假性血友病因子的变化。结果 :治疗前 ,心房纤颤患者血小板P选择素、血浆血管性假性血友病因子显著增高。经葛根素与阿斯匹林治疗均可降低P选择素、血浆血管性假性血友病因子水平。且葛根素和阿斯匹林组疗效无明显差异。结论 :心房纤颤患者存在血小板活性增强和内皮功能受损 ,葛根素可显著改善血小板活化状态和内皮细胞功能 ,且能与阿斯匹林相媲美     

葛根素口腔崩解片的研制

目的：以葛根素为模型药物制备口腔崩解片。方法该研制把崩解时间及沉降容积比为指标,单因素法筛选片剂的处方组成及工艺,并优化制备工艺。结果葛根素口腔崩解片的辅料为甘露醇、明胶、阿司帕坦与薄荷香精,经过冷冻干燥方法制备,口感良好,4s的崩解时间,在4min内体外溶出度达96．46％。结论葛根素口腔崩解片可迅速崩解于口腔内,制备工艺可行。     

葛根素联合生脉注射液治疗小儿病毒性心肌炎的疗效分析

目的观察葛根素联合生脉注射液治疗小儿病毒性心肌炎的临床疗效。方法将66例小儿病毒性心肌炎患者随机分为3组。3组均给予常规治疗,治疗组Ⅰ在常规治疗的基础上加用生脉注射液,治疗组Ⅱ在治疗组Ⅰ的基础上加用葛根素注射液。观察治疗前后症状、体征、心电图、心肌酶谱等指标的变化。结果治疗组Ⅰ在各项指标上明显优于对照组（P〈0.05）。治疗组Ⅱ与治疗组Ⅰ相比,各项指标的改善情况更为明显（P〈0.05）。结论生脉注射液治疗小儿病毒性心肌炎疗效显著。葛根素联合生脉注射液治疗疗效更佳,值得临床推广。     

Effects of particle size on the pharmacokinetics of puerarin nanocrystals and microcrystals after oral administration to rat

Puerarin, which is extracted from traditional Chinese medicine, is widely used in clinic in China and mainly used as a therapeutic agent to cardiovascular diseases. Owing to its poor water solubility and adverse drug reactions caused by cosolvents after intravenous administration, the development of oral formulation is urgently needed. Nowadays, nanocrystals technique has become a preferred way to develop oral dosage form. In this study, we used high pressure homogenization (HPH) to prepare puerarin nanocrystals and microcrystals with different sizes ranged from 525.8 nm to 1875.6 nm and investigated the influence of particle size on pharmacokinetics. The nanocrystals and microcrystals prepared were characterized using DLS, DSC, XRD and SEM, and we found that the crystalline state of puerarin was changed during the preparation process and the drug was dispersed into HPMC. In the pharmacokinetic study, we observed an increasing of C_max and AUC and a decreasing of CL/F with the decreasing of particle size. The AUC of the puerarin nanocrystals (525.8 nm) was 7.6-fold of that of raw puerarin suspension, with an absolute bioavailability of 21.44%. From the above results, we can conclude that nanocrystal technique is an efficient technology to improve the oral bioavailability of puerarin.