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.     

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。     

葛根素对难治性心力衰竭患者氧化/抗氧化系统失衡的调节作用

目的：探讨葛根素对难治性心力衰竭患者血中氧化/抗氧化指标的调节作用。方法：选取健康成人34例，检测静脉血全血超氧 化物歧化酶(SOD)、全血谷胱甘肽过氧化物酶(GSH-Px)和过氧化氢酶(CAT)的活性及血浆脂质过氧化物(LPO)和丙二醛(MDA)的 浓度，然后与难治性心力衰竭组治疗前的此5项指标进行比较；选择76例难治性心力衰竭患者，随机分成两组，试验前均监测静脉血 全血 SOD、GSH-Px 和 CAT 的活性及血浆 LPO 和 MDA 的浓度并评价心功能，然后试验组在给予常规治疗的同时合并给予葛根素 注射液300 mg/d，稀释后静脉滴注，疗程2周；对照组只给予常规治疗，2周后，复测上述指标，比较各组治疗前后的变化。结果：1) 难 治性心力衰竭患者全血 SOD、全血 GSH-Px 和 CAT 的活性比健康对照组低，而血浆 LPO 及 MDA 的浓度比健康对照组高，均 P＜0． 01；2) 葛根素组试验前后比较，全血 SOD、全血 GSH-Px 和 CAT 活性明显增高，且血浆 LPO 和全血 MDA 浓度明显降低，均P＜0． 05；而对照组则无显著变化，P＞0． 05；3) 葛根素组心力衰竭纠正的有效率显著高于对照组，P＞0． 05。结论：难治性心力衰竭患者机 体的抗氧化防御能力降低，葛根素可通过提高机体的抗氧化能力，消除氧自由基和脂质过氧化物，起到纠正心力衰竭的作用。     

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。     

葛根素降低糖尿病大鼠血清糖基化终产物水平和单核细胞趋化蛋白1含量

目的　研究葛根素对糖尿病大鼠糖基化终产物 (AGEs)水平和单核细胞趋化蛋白 1(MCP 1)表达的影响。方法　将大鼠随机分为正常对照组 (CON)、糖尿病组 (DM )、糖尿病氨基胍治疗组 (AG )和糖尿病葛根素治疗组 (PU )。腹腔注射链脲佐菌素诱导糖尿病模型 ,检测血清中AGEs(荧光法 )和MCP 1(ELISA)水平 ;病理切片PAS染色及电镜观察肾组织的病理改变。免疫组化检测肾皮质MCP 1的蛋白表达水平。结果　DM组血清AGEs和MCP 1水平较PU组和AG组明显增高 (均P <0 .0 5 ) ;电镜发现两个治疗组的肾小球基底膜和心肌细胞病理改变程度比糖尿病组轻 ;肾小球内皮细胞和系膜区MCP 1免疫组化染色阳性细胞数明显比糖尿病组少。结论　葛根素能降低糖尿病大鼠血清AGEs和MCP 1水平 ,减少肾皮质中MCP 1的表达 ,减轻肾脏和心肌的病变程度     

Kudzu extract treatment does not increase the intoxicating effects of acute alcohol in human volunteers

Background: Isoflavone administration in the form of a purified extract from the herbal medication kudzu root has been shown to reduce, but not eliminate, alcohol consumption in alcohol‐abusing and alcohol‐dependent men. The precise mechanism of this action is unknown, but 1 possible explanation for these results is that the isoflavones in kudzu might actually increase the intensity or duration of alcohol’s effects and thus delay the desire for subsequent drinks. This study was designed to test this hypothesis. Methods: Twelve (12) healthy adult men and women (27.5 ± 1.89 years old) who consumed moderate amounts of alcohol (7.8 ± 0.63 drinks/wk) participated in a double‐blinded, placebo‐controlled crossover study in which they were treated with either kudzu extract (total isoflavone dose of 750 mg/d) or matched placebo for 9 days. On days 8 and 9, participants received an acute challenge of ethyl alcohol (either 0.35 or 0.7 g/kg alcohol). During the challenges, the following measures were collected: subjective effects, psychomotor (body sway), cognitive performance (vigilance/reaction time), physiological measures (heart rate and skin temperature), and plasma ethanol concentration. Results: Alcohol resulted in a dose‐related alteration in subjective measures of intoxication, impairment of stance stability, and vigilance/reaction time. Kudzu extract did not alter participants’ subjective responses to the alcohol challenge or to alcohol’s effects on stance stability or vigilance/reaction time. However, individuals treated with kudzu extract experienced a slightly more rapid rise in plasma ethanol levels, but only after the 0.7 g/kg dose. This transient effect during the first 30 minutes of the ascending plasma alcohol curve lasted only 10–15 minutes; there were no differences in peak plasma alcohol levels or alcohol elimination kinetics. Additionally, kudzu pretreatment enhanced the effects of the 0.7 g/kg dose of alcohol on heart rate and skin temperature. Conclusions: These data suggest that individuals who drink alcohol while being treated with kudzu extract experience no adverse consequences, and furthermore the reported reductions in alcohol intake after kudzu extract treatment are not related to an alteration in alcohol’s subjective or psychomotor effects.     

丹参联合葛根对糖尿病眼病模型大鼠的协同作用

目的:研究丹参联合葛根对糖尿病眼病模型大鼠的协同作用。方法:腹腔注射链脲佐菌素-弗氏完全佐剂联合喂饲高脂饲料以复制大鼠糖尿病眼病模型。实验分为正常对照(等容生理盐水)组、模型(等容生理盐水)组、葛根(33 g/kg)组、葛根丹参(33 g/kg+33 g/kg)组,灌胃给药,每天2次,连续15 d。于电镜下观察大鼠眼组织形态变化。分别灌胃葛根(33 g/kg)、葛根丹参(33g/kg+33 g/kg),给药0、10、20、30、45、60、90、120、240、360、480 min时取血,高效液相色谱法测定大鼠血浆中葛根素的血药浓度。色谱柱为Diamonsil C18(250 mm×4.6 mm,5μm),流动相为甲醇-水(25∶75,V/V),检测波长为250 nm,流速为1.0 ml/min,柱温为25Ⅸ,进样量为10μl。以WinNonlin软件分析药动学参数。结果:模型组大鼠眼组织视网膜细胞萎缩,脉络膜血管增生、扩张、充血,出现新生血芽,内皮细胞核数目增多,给药后上述病理状态均得到改善,其中葛根丹参组改善情况优于葛根组。葛根素质量浓度在0.1⁸.0 mg/ml范围内与其同内标峰面积比值呈良好线性关系,精密度试验RSD均小于3%,稳定性试验RSD均小于4%,回收率在86.02%8.0 mg/ml范围内与其同内标峰面积比值呈良好线性关系,精密度试验RSD均小于3%,稳定性试验RSD均小于4%,回收率在86.02%⁹0.28%。葛根组与葛根丹参组AUC分别为(156.49±32.62)、(462.27±119.86)mg·min/L,cmax分别为1.426 5、1.986 2 mg/ml,t1/2kα分别为(9.62±1.59)、(10.07±2.11)min。结论:丹参与葛根配伍,可明显促进葛根素的体内吸收、分布,减缓其消除,促进葛根素在血液中的富集且能改善模型大鼠眼组织病变。丹参对葛根改善模型大鼠糖尿病眼病具有较好协同增效作用。     

主动脉阻断后经主动脉根部注射葛根素注射液对二尖瓣置换术患者心肌损伤的影响

目的：评价主动脉阻断后经主动脉根部注射葛根素注射液对二尖瓣置换术患者心肌损伤的影响。方法：拟行二尖瓣置换术患者60例,性别不限,年龄〈60岁,ASA分级Ⅱ或Ⅲ级,采用随机数字表法,将患者随机分为C组和P组,每组30例。在体外循环下,P组于阻断升主动脉后,立即经升主动脉根部注射葛根素注射液5mg/kg,随后灌注4℃高钾（K＋浓度22 mmol/L）含血心脏停搏液20 mL/kg;C组注射等容量生理盐水,随后灌注4℃高钾（K＋浓度22 mmol/L）含血心脏停搏液20 mL/kg,两组患者每30 min复灌不含葛根素的相同成分含血心脏停搏液10 mL/kg。分别于主动脉阻断前即刻（T1）、主动脉阻断后4 h（T2）和术后24 h（T3）时,采集中心静脉血样8 mL,测定血浆心肌肌钙蛋白I（cTnI）浓度、肌酸激酶（CK）和肌酸激酶同功酶（CK-MB）的活性。记录阻断升主动脉至心脏停搏所用时间、心脏自动复跳率、心脏复跳期间利多卡因使用剂量及术后2h多巴胺用量。结果：与C组比较,P组心脏复跳期间利多卡因用量和术后2 h多巴胺用量减少,两组比较差异均有统计学意义（P〈0.05）。与C组比较,P组T2~T3时血浆cTnI、CK和CK-MB的活性降低,两组比较差异均有统计学意义（P〈0.05）。结论：主动脉阻断后经主动脉根部注射葛根素注射液可减轻二尖瓣置换术患者心肌损伤。     

津力糖平片对照品溶液稳定性的研究

目的：探讨津力糖平片检验用葛根素对照品溶液的保存条件和有效期,规范中药检验对照品的管理。方法：按照《中华人民共和国药典：一部》中药的薄层色谱法对葛根素对照品溶液进行薄层成像实验,比较不同时间、不同保存条件下不同质量浓度对照品溶液的成像效果。结果：葛根素溶液最佳保存时间为配制后270d内,高浓度对照品溶液（10mg/ml）成像效果更好,常温和冷藏（2—8℃）保存对实验结果无影响。结论：本研究为中药对照品保存期限设定和标准化管理提供了一定依据,有利于减少葛根素对照品的损耗、节约药品检验成本。     

葛根素联合甲钴胺治疗糖尿病周围神经病变的有效性和安全性

目的 探讨葛根素注射液联合甲钴胺注射液治疗糖尿病周围神经病变（DPN）的有效性和安全性。方法将110例DPN患者随机分为2组,对照组55例给予甲钴胺注射液静脉注射治疗,在对照组基础上,治疗组给予葛根素注射液静脉注射治疗,疗程4周。观察两组治疗前后总神经症状积分（TSS）、感觉神经传导速（SNCV）和运动神经传导速（MNCV）,同时比较两组治疗前后各指标的变化。结果 两组治疗两周后,治疗组总有效率为89.1%,对照组总有效率为76.4%,两组治疗结果比较有统计学意义（P〈0.05）;TSS较治疗前明显降低,SNCV和MNCV显著提高,两组各指标治疗前后比较,P〈0.05,而且组间比较差异均有统计学意义（P〈0.05）。结论葛根素注射液联合甲钴胺注射液能缓解糖尿病周围神经病变患者临床症状,提高神经传导速度,有显著的临床疗效。