积雪草苷平衡溶解度和油水分配系数的测定

目的测定积雪草苷在不同pH介质中的平衡溶解度和正辛醇-水/缓冲液体系中的表观油水分配系数（Papp）。方法采用RP-HPLC法测定积雪草苷浓度,研究其在水和不同pH介质中的平衡溶解度;采用摇瓶法测定积雪草苷在正辛醇-水/缓冲液体系中的Papp。结果 25℃下积雪草苷在水中的平衡溶解度为305.02μg.ml-1,在pH 5.0缓冲液中平衡溶解度更低;积雪草苷在正辛醇-水中Papp为0.908（logPapp=-0.042）,在pH 5.0缓冲液中Papp最高。结论积雪草苷为亲水性药物,但其在水中的溶解度较小。在pH 5.0～8.0范围内,积雪草苷的平衡溶解度随pH的增加而增加,相反油水分配系数随pH的增加而减少。     

积雪苷软膏对BALB/C裸小鼠瘢痕疙瘩模型成纤维细胞及胶原蛋白表达的影响

目的：明确外用积雪苷软膏对瘢痕疙瘩裸鼠模型的成纤维细胞和胶原蛋白表达的影响。方法：取手术切除的瘢痕疙瘩组织植入16只裸鼠肩胛及骶尾部，建立瘢痕疙瘩裸鼠模型。随机分为A组（外用对照软膏基质），B组（外用0.00125%积雪苷软膏），C组 （外用0.0025%积雪苷软膏）和D组（外用0.005%积雪苷软膏），每组4只。观察瘢痕疙瘩大体形态学及光镜下变化；免疫组化检测瘢痕疙瘩I型胶原蛋白表达的影响；透射电镜观察瘢痕疙瘩成纤维细胞结构的变化。结果：大体观察裸鼠瘢痕疙瘩经不同浓度积雪苷软膏作用后组织块体积均变小，光镜观察成纤维细胞数明显减少。A～D组I型胶原蛋白的表达率分别为79.17%、70.83%、37.50％、33.33%，四组比较，差异具有统计学意义（Fisher精确概率检验法，P<0.05）。透射电镜观察B-D组瘢痕疙瘩成纤维细胞较A组明显减少，可见凋亡细胞，细胞核固缩，膜结构不清，有坏死的细胞碎片及裸核状态的成纤维细胞。结论：外用积雪苷软膏能减少瘢痕疙瘩成纤维细胞并下调I型胶原蛋白的表达。     

载积雪草总苷同轴和混纺纳米纤维膜的制备和缓释性能评价

目的 采用静电纺丝法制备同轴及混纺载积雪草总苷的纳米纤维膜,研究其形貌及体外释药行为,为深Ⅱ度烧伤提供新型功能性敷料.方法 采用静电纺丝工艺,制备芯层载积雪草总苷的同轴海藻酸钠／聚乙烯醇-壳聚糖纳米纤维膜.进行处方工艺优化,包括壳层、芯层中聚合物浓度、制备工艺等,同时制备海藻酸钠／聚乙烯醇混纺载积雪草总苷纳米纤维膜.用扫描电子显微镜、透射电子显微镜和X射线衍射表征纳米纤维.考察载积雪草总苷混纺和同轴纳米纤维膜的体外释药行为.结果 载积雪草总苷同轴纳米纤维膜的最优处方是壳层由0.8％海藻酸钠、7％聚乙烯醇混合溶液制备,芯层由含2.5％积雪草苷的3％壳聚糖溶液制备.最优工艺是静电电压23 kV,针头和接收屏距离15 cm,壳层溶液推注速度0.8 ml/h,芯层溶液推注速度0.2 ml/h.混纺纳米纤维膜与同轴纳米纤维膜的处方和工艺类似.同轴纳米纤维膜有明显壳核结构,其芯层直径为99.2nm,整个纤维直径为166.8 nm.与混纺纳米纤维比较,同轴纳米纤维膜有药物缓释效果,机制以Fick's扩散为主.结论 同轴纳米纤维膜有明显药物缓释效果,有利于积雪草苷发挥促愈合效果,可用于深Ⅱ度烧伤的治疗.     

积雪苷霜软膏治疗黄褐斑随机、双盲、自身对照试验研究

目的：评估积雪苷霜软膏R治疗黄褐斑的临床疗效及安全性。方法：采用12周随机、双盲、自身对照试验,受试者左右两侧面部黄褐斑区域随机使用积雪苷霜软膏R与不含积雪草总苷成分的安慰剂,观察色斑改善程度。通过第0周、第4周、第8周、第12周研究者盲法评价黄褐斑面积及严重程度指数（MASI）、受试者盲法自评、仪器测量三个方面评价比较用药侧与安慰剂侧黄褐斑的改善情况。结果：共纳入符合要求的研究对象36例全部按期完成,在试验结束揭盲后统计,受试者的治疗侧MASI指数下降45%,结合受试者对疗效的自我评估、颜色定量测量及拍照,治疗侧疗效明显优于安慰剂侧。结论：本研究结果显示积雪苷霜软膏R治疗黄褐斑皮肤耐受性良好,临床疗效明显优于安慰剂。     

皮肤磨削及术后外用硅凝胶或积雪苷治疗面部增生性瘢痕的疗效比较

目的：比较单纯皮肤磨削以及磨削术后联合外用硅凝胶或积雪苷治疗面部增生性瘢痕的临床疗效。方法：皮肤磨削术后外用莫匹罗星软膏,将101例随机分为三组：A组为对照组;B组和C组为治疗组,分别于术后5天起外用硅凝胶和0.25%积雪苷霜至术后8周结束。均观察治疗结束时红斑及色素沉着发生率,治疗前后瘢痕严重程度的积分评价。结果：与对照组A比较,治疗C组红斑发生率及色素沉着发生率均降低,C组的色素沉着发生率低于B组;B组和C组的瘢痕疗效明显优于A组,而B组和C组之间差异无显著性。结论：皮肤磨削术后联合外用硅凝胶或者积雪苷治疗面部增生性瘢痕效果可靠,后者还可以较好地改善患者肤色。     

Antipyretic and Anti‐inflammatory Effects of Asiaticoside in Lipopolysaccharide‐treated Rat through Up‐regulation of Heme Oxygenase‐1

Asiaticoside (AS), a triterpenoid isolated from Centella asiatica, has been found to exhibit antioxidant and anti‐inflammatory activities in several experimental animal models. However, the underlying mechanisms remain elusive. In this study, we provide experimental evidences that AS dose‐dependently inhibited lipopolysaccharide (LPS)‐induced fever and inflammatory response, including serum tumor necrosis factor (TNF)‐α and interleukin (IL)‐6 production, liver myeloperoxidase (MPO) activity, brain cyclooxygenase‐2 (COX‐2) protein expression and prostaglandin E₂ (PGE₂) production. Interestingly, AS increased serum IL‐10 level, liver heme oxygenase‐1 (HO‐1) protein expression and activity. Furthermore, we found that the suppressive effects of AS on LPS‐induced fever and inflammation were reversed by pretreatment with ZnPPIX, a HO‐1 activity inhibitor. In summary, our results suggest that AS has the antipyretic and anti‐inflammatory effects in LPS‐treated rat. These effects could be associated with the inhibition of pro‐inflammatory mediators, including TNF‐α and IL‐6 levels, COX‐2 expression and PGE₂ production, as well as MPO activity, which might be mediated by the up‐regulation of HO‐1. Copyright © 2012 John Wiley & Sons, Ltd.     

积雪苷口腔崩解片的研制及质量控制

目的研制积雪苷口腔崩解片。方法以积雪苷为主药,内加淀粉为崩解剂,建立性状、崩解时限、含量测定等质控方法。结果制备的口腔崩解片符合工艺要求。以高效液相色谱法测定积雪苷口腔崩解片中积雪草苷的含量,平均回收率为100.7%,RSD=1.01%（n=9）。结论该制剂制备工艺可行,测定方法简便、准确。     

手性流动相添加剂法测定积雪草中积雪草总苷的量及其总苷特征图谱

目的 以β-环糊精为流动相添加剂,拆分积雪草药材及其总苷中羟基积雪草苷和积雪草苷,建立羟基积雪草苷和积雪草苷的定量测定方法,同时建立积雪草总苷的高效液相色谱特征图谱。方法 采用 Dikma Diamonsil^TM C₁₈ (250 mm×4.6 mm, 5 μm) 色谱柱;流动相为乙腈-2 mmol/L β-环糊精溶液 (24∶76);柱温 30 ℃;体积流量 1.0 mL/min;检测波长 205 nm;理论塔板数按积雪草苷峰计算应不低于 4 000。结果 β-环糊精的加入很好地分离积雪草药材及其总苷中的羟基积雪草苷和积雪草苷,满足了定量分析的要求,其中有效成分羟基积雪草苷和积雪草苷的线性范围分别为 0.181～18.1 μg (r=0.999 98) 和 0.195～19.5 μg (r=0.999 98) ,积雪草药材的平均加样回收率 (n=9) 分别为 101.2% 和 101.6%,积雪草总苷的平均加样回收率 (n=9) 分别为 100.3% 和 99.3%,11 批次的积雪草总苷样品的高效液相色谱特征图谱中确定 6 个共有峰作为定性的指标峰。结论 本法简便快速、结果准确、重现性好,可用于积雪草药材及其总苷的质量控制。     

Induction of keratinocyte migration by ECa 233 is mediated through FAK/Akt,ERK, and p38 MAPK signaling

Centella asiatica is widely considered the most important medicinal plant for treating and relieving skin diseases. Recently developed standardized extract of Centella asiatica ECa 233 has demonstrated positive effects on wound healing of incision and burn wound in rats. However, knowledge associated with wound healing mechanism of ECa 233 was scare. Therefore, this study aimed to investigate the effect and underlying molecular mechanisms of ECa 233 on the migration of a human keratinocyte cell line (HaCaT) using scratch wound healing assay. Formation of filopodia, a key protein in cell migration as well as signaling pathways possibly involved were subsequently assessed. It was found that HaCaT cell migration was significantly enhanced by ECa 233 in a concentration‐ and time‐dependent manner. The filopodia formations were accordingly increased in exposure to ECa 233 at concentrations of 0.1–100 μg/ml. Furthermore, ECa 233 was found to significantly upregulate the expression of Rac1 and RhoA and to induce phosphorylation of FAK and Akt as well as ERK and p38 MAPK. Taken all together, it is suggestive that ECa 233 induces cell migration and subsequently promotes wound healing activity, through the activation of FAK, Akt, and MAPK signaling pathways thereby supporting the role of ECa 233 to be further developed for the clinical treatment of wound.     

积雪草苷对DBA/1小鼠胶原诱导型关节炎中Th17/Treg细胞表达的影响

目的:观察积雪草苷对DBA/1小鼠胶原诱导型关节炎(CIA)中辅助性T细胞17(Th17)和调节性T细胞(Treg)表达的影响.方法:将SPF级DBA/1雄性小鼠按体质量随机分为6组:正常组,CIA组,甲氨蝶呤组(MTX组,0.5 mg·kg-1),积雪草苷低、中、高剂量(5,15,45 mg· kg-1)组.除正常组...