Comparative Study on Transdermal Absorption of Bulleyaconitine A in Four Skin Barrier Models of Rabbits
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摘要:
目的 对比研究草乌甲素凝胶贴在正常家兔正常皮肤、正常家兔破损皮肤、病理损伤正常皮肤、病理损伤破损皮肤4种皮肤屏障模型在体透皮吸收。 方法 采用肌肉注射50%甘油的方法获得病理损伤模型家兔，采用1号砂纸包裹木棒轻轻摩擦皮肤表面至变为浅红的方法制备破损皮肤。4种皮肤屏障模型经皮给予草乌甲素凝胶贴，正常皮肤于给药后1 h、2 h、3 h、4 h、5 h、6 h、8 h、10 h采血，破损皮肤于给药后1 h、2 h、3 h、4 h、5 h、6 h、8 h、10 h、12 h、24 h、36 h采血，HPLC-MS/MS检测4种模型家兔草乌甲素给药后不同时间点血浆药物浓度，采用DAS 3.2.0计算药代参数，SPSS 23.0进行组间药代参数统计。 结果 AUC(0-t)、AUC(0-∞)、Cmax 3个药代参数在4种皮肤屏障模型家兔的变化趋势一致，大小分别为：正常家兔破损皮肤>病理损伤破损皮肤>病理损伤正常皮肤>正常家兔正常皮肤。皮肤外环境不同对草乌甲素透过率产生较大影响，破损皮肤透过率明显高于正常皮肤（P < 0.05）；单纯病理损伤（内伤），皮肤状态一致，草乌甲素的吸收、消除的程度和速度均无明显影响（P > 0.05）；模拟外伤所致正常家兔破损皮肤、内外伤的病理损伤破损皮肤与正常家兔正常皮肤相比，草乌甲素吸收程度大幅提高、消除能力明显降低（P < 0.05）。 结论 皮肤角质层的屏障功能是草乌甲素透皮吸收的主要限制因素，外伤或内外伤状态下，可能导致草乌甲素在体内蓄积，影响用药安全。在进行经皮给药制剂试验研究时，应同时进行正常和模拟外伤所致的破损皮肤或内外伤的病理损伤破损皮肤模型对比研究，模拟临床肌肉、皮肤受到内外伤的情况下透皮吸收，将有助于更加全面准确地评价经皮给药制剂药效、药代和毒理特性。 Abstract:Objective To compare the transdermal absorption of Bulleyaconitine A on normal rabbit intact skin, normal rabbit damaged skin, pathological damaged normal skin, and pathological damaged injured skin barrier models in vivo. Methods The rabbit model of pathological injury was obtained by intramuscular injection of 50% glycerol. The damaged skin was prepared by gently rubbing the skin surface with a wooden stick wrapped in No. 1 sandpaper until it turned light red. Four skin barrier models were topically treated with Bulleyaconitine A gel. Blood samples were collected at various time points after drug administration for normal skin （1 h, 2 h, 3 h, 4 h, 5 h, 6 h, 8 h, 10 h） and damaged skin （1 h, 2 h, 3 h, 4 h, 5 h, 6 h, 8 h, 10 h, 12 h, 24 h, 36 h）. HPLC-MS/MS was used to detect drug concentrations in rabbit models at different time points after Carbenoxolone administration. Drug pharmacokinetic parameters were calculated using DAS 3.2.0, and inter-group pharmacokinetic parameters were statistically analyzed using SPSS 23.0. Results The trends of three pharmacokinetic parameters, AUC（0-t）, AUC（0-∞）, and Cmax, in four skin barrier models of rabbits are consistent. They are as follows in size order: intact skin of normal rabbits > damaged skin from pathological injury > intact skin from pathological injury > intact skin of normal rabbits. Different external skin environments have a significant impact on the permeability of Bulleyaconitine A absorption. The permeability of damaged skin is significantly higher than that of normal skin （P < 0.05）. Pure pathological damage （internal injury）, with consistent skin conditions, shows no significant impact on the absorption and elimination of Chuanwu （P > 0.05）. Compared to intact skin of normal rabbits, damaged skin from simulated external trauma, and damaged skin from internal and external trauma, the absorption of Bulleyaconitine A significantly increases and the elimination capacity significantly decreases （P < 0.05）. Conclusion The barrier function of the stratum corneum is the main limiting factor for Bulleyaconitine A transdermal absorption. Bulleconitine A may accumulate in the body under the condition of trauma or internal or external injuries, which may affect drug safety. When conducting experiments on transdermal drug delivery formulations, it is advisable to simultaneously compare normal skin with damaged skin caused by simulated trauma or pathological damage from internal and external injuries. Studying the transdermal absorption under conditions simulating clinical muscle and skin injuries from internal and external trauma will help in a more comprehensive and accurate evaluation of the efficacy, pharmacokinetics, and toxicological properties of transdermal drug delivery formulations. -
Key words:
- Bulleyaconitine A /
- Normal skin /
- Damaged skin /
- Pathological injury /
- HPLC-MS/MS
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图 3 专属性考察图
A：空白兔血浆；B：草乌甲素-血浆加标；C：次乌头碱-血浆加标；D：草乌甲素-含药血浆。
Figure 3. Investingation of method specificity
图 4 物理挫伤病理检查（×10）
A：出血、肌纤维坏死；B：出血；C：出血、水肿、肌纤维坏死；D：肉芽组织、瘢痕形成。
Figure 4. Pathological examination of physical contusion（×10）
图 5 化学损伤病理检查（×10）
A：出血、肌纤维坏死；B：水肿、肌纤维坏死；C：肌纤维坏死、瘢痕形成；D：出血、肌纤维坏死、肉芽组织形成。
Figure 5. Pathological examination of chemical damage（×10）
图 6 草乌甲素在4种不同皮肤模型家兔体内平均药-时曲线
Figure 6. The average plasma concentration-time curves of Bulleyaconitine A in rabbits with four different skin models
表 1 LC-MS/MS测定兔血浆中草乌甲素的精密度、准确度和回收率[（$ \bar x \pm s$），n = 6]
Table 1. Determination of precision，accuracy and recovery of Bulleyaconitin A in rabbit plasma by LC-MS/MS [（$ \bar x \pm s $），n = 6]
质控浓度
（ng/mL）日内（n=6） 日间（n=18） 回收率
（n=6）（%）（ng/mL） 精密度（%） 准确度（%） （ng/mL） 精密度（%） 准确度（%） 0.2050 0.2070±0.01000 4.83 100.98 0.2002±0.0179 8.94 97.66 81.74±1.67 3.275 3.024±0.266 8.80 92.34 3.086±0.255 8.26 94.23 82.11±2.88 52.50 50.98±2.45 4.80 97.10 54.39±4.14 7.61 103.60 87.21±2.55 
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表 2 4种不同皮肤模型家兔体内主要药动学参数[（$ \bar x \pm s$），n = 6]
Table 2. Main pharmacokinetic parameters of Bulleyaconitine A in rabbits with four different skin models [（$ \bar x \pm s$），n = 6]
参数 正常家兔正常皮肤 正常家兔破损皮肤 病理损伤正常皮肤 病理损伤破损皮肤 AUC（0-t）（μg/L·h） 2.199±2.204 25.117±16.462 2.682±1.834 20.834±11.167 AUC（0-∞）（μg/L·h） 3.215±4.099 25.637±16.244 3.059±1.934 21.553±10.376 Cmax（μg/L） 0.379±0.316 2.495±1.570 0.472±0.302 1.655±0.828 Tmax（h） 3.667±1.033 3.5±1.378 3.167±0.983 4.833±1.472 t1/2z（h） 3.573±2.725 9.533±10.476 3.071±0.885 13.244±22.289 CL（L/h） 3882.402±3555.427 362.47±446.772 1709.041±849.386 251.724±192.713
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表 3 4种皮肤模型家兔体内主要药动学参数显著性差异统计结果
Table 3. Statistical results of significant differences about main pharmacokinetic parameters in rabbits with four different skin models
药动参数
显著性差异正常家兔正常皮肤-
正常家兔破损皮肤病理损伤正常皮肤-
病理损伤破损皮肤正常家兔正常皮肤-
病理损伤正常皮肤正常家兔破损皮肤-
病理损伤破损皮肤正常家兔正常皮肤-
病理损伤破损皮肤P[AUC（0-t）（μg/L·h）] <0.016* <0.003* <0.522 <0.609 <0.02* P[AUC（0-∞）（μg/L·h）] <0.010* <0.002* <0.522 <0.615 <0.02* P[Cmax（μg/L）] <0.025* <0.025* <0.522 <0.273 <0.016* P[Tmax（h）] <0.817 <0.044* <0.411 <0.136 <0.143 P[t1/2z（h）] <0.078 <0.055 <0.749 <0.337 <0.150 P[CL（L/h）] <0.010* <0.006* <0.522 <0.423 <0.006* *P < 0.05。
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[1] Xie M X,Zhu H Q,Pang R P,et al. Mechanisms for therapeutic effect of Bulleyaconitine A on chronic pain[J]. Molecular Pain,2018,14:1-10. [2] Zhu H Q,Xu J,Shen K F,et al. Bulleyaconitine A depresses neuropathic pain and potentiation at C-fiber synapses in spinal dorsal hom induced by paclitaxel in rats[J]. Exp Neurol,2015,273:263-272. doi: 10.1016/j.expneurol.2015.09.006 [3] 刘延青,丁晓宁,王应德. 草乌甲素片治疗常见慢性疼痛的临床研究[J]. 中国疼痛医学杂志,2011,17(5):314-315. doi: 10.3969/j.issn.1006-9852.2011.05.016 [4] Xu L,Lao LX,Ge A,et al. Chinese herbal medicine for cancer pain[J]. Intergrative Cancer Therapies,2007,6(3):208-234. doi: 10.1177/1534735407305705 [5] 吴玲普. 草乌甲素对膝关节骨折术后运动疼痛的干预[D]. 广州: 广州中医药大学, 2010. [6] 王薇. 北京市癌痛治疗现状调查分析及草乌甲素治疗轻中度癌痛的临床研究[D]. 北京: 中国中医科学院, 2019. [7] 牛占国,陈晓武,黄静,等. 草乌甲素片用于烧伤创面镇痛效果的临床观察[J]. 吉林医学,2014,35(31):7001-7002. doi: 10.3969/j.issn.1004-0412.2014.31.072 [8] 李芳,倪家骧,武百山,等. 草乌甲素胶丸辅助硫酸吗啡缓释片治疗中晚期癌痛[J]. 长春中医药大学学报,2014,30(3):460-462. [9] Wang C F,Gerner P,Schemidt B,et al. Use of Bulleyaconitine A as an adjuvant for prolonged cutaneous analgesia in the rat[J]. Anesthesia& Analgesia,2018,107(4):1397-1405. [10] Peng J,Xiao S,Xie J,et al. Bulleyaconitine A reduces fracture-induced pain and promotes fracture healing in mice[J]. Front Pharmacol.,2023,14:1046514. doi: 10.3389/fphar.2023.1046514 [11] 陈新鹏,陈娟,曾惠琼,等. 草乌甲素片联合甲氨喋呤治疗类风湿关节炎的临床疗效[J]. 临床医学,2018,38(5):116-118. [12] Zhan X D,Zhang W Q,Sun T,et al. Bulleyaconitine A effectively relieves allergic lung inflammation in a murine asthmatic model[J]. Medical Science Monitor:international Medical Journal of Experimental and Clinical Research,2019,25:1656-1662. [13] 张文琪. 草乌甲素对过敏性哮喘小鼠肺部炎症的治疗与机理初步探讨[D]. 芜湖: 皖南医学院, 2019. [14] 孙甜,吕业超,唐小牛,等. 草乌甲素对LPS诱导的RAW264.7细胞抗炎作用及相关机制研究[J]. 泰山医学院学报,2020,41(5):321-327. [15] Liu L P,Wang S Y,Xing H Y,et al. Bulleyaconitine a inhibits the lung inflammation and airway remodeling through restoring Th1/Th2 balance in asthmatic model mice[J]. Bioscience,Biotechnology,and Biochemistry,2020,84(7):1409-1417. [16] 陈建洪,邓莎,吴志云,等. 草乌甲素胶丸治疗肩周炎的疗效观察[J]. 临床合理用药杂志,2012,5(26):30. doi: 10.3969/j.issn.1674-3296.2012.26.021 [17] 潘国兴,杜冬萍. 草乌甲素联合玻璃酸钠关节腔内注射治疗膝关节骨性关节炎的疗效[J]. 上海医学,2013,36(6):523-526. [18] 李冬冬,殷娜,王志伟. 草乌甲素片联合筋骨痛消丸治疗早中期膝骨关节炎45例临床观察[J]. 风湿病与关节炎,2019,8(6):27-30. doi: 10.3969/j.issn.2095-4174.2019.06.007 [19] Weng W Y,Xu H N,Huang J M,et al. A pharmacokinetic study of intramuscular administration of Bulleyaconitine A in healthy volunteers[J]. Biological& Pharmaceutical Bulletin,2005,28(4):747-749. [20] 肖若兰. 草乌甲素多囊脂质体和聚合物胶束给药系统的研究[D]. 沈阳: 沈阳药科大学, 2008. [21] 吴晓辉. 草乌甲素微乳及其水性基质经皮给药系统研究[D]. 北京: 中国协和医科大学, 2006. [22] 张伟,张玲,杨兆祥. HPLC法测定草乌甲素贴释放度[J]. 云南中医学院学报,2010,33(3):21-24,29. doi: 10.3969/j.issn.1000-2723.2010.03.008 [23] 翁伟宇. 草乌甲素经皮渗透特性及其透皮给药系统的研究[D]. 上海: 复旦大学, 2004. [24] 廖淑彬,陈丹,余文静,等. 三叶青地上部分抗炎提取物效应组分群不同生理状态体内组织分布规律及特性研究[J]. 中国中药杂志,2021,46(5):1224-1249. [25] 郝晶晶,郭瑛玉,胡海燕,等. 金红片中活性成分在正常大鼠和浅表性胃炎大鼠体内的药代动力学研究[J]. 药物分析杂志,2023,43(9):1565-1573. [26] 刘莹,贾兰,张晓喻,等. 大黄素配伍黄芩素在正常和2型糖尿病大鼠中药代动力学研究[J]. 中国临床药理学杂志,2023,39(19):2834-2838. [27] Liu F,Nong X,Qu W,Li X. Pharmacokinetics and tissue distribution of 12 major active components in normal and chronic gastritis rats after oral administration of Weikangling capsules[J]. Journal of Ethnopharmacol,2023,316:116722. doi: 10.1016/j.jep.2023.116722 [28] Zhou H,He Y,Zheng Z,et al. Pharmacokinetics and tissue distribution study of 18 bioactive components in healthy and chronic heart failure rats after oral administration of Qi-Shen-Ke-Li formula using ultra-high-performance liquid chromatography/triple quadrupole mass spectrometry[J]. Rapid Commun Mass Spectrom,2021,35(8):9060. doi: 10.1002/rcm.9060 [29] Mei Y D,Tong X Y,Hu Y M,et al. Comparative pharmacokinetics of six bioactive components of Shen-Wu-Yi-Shen tablets in normal and chronic renal failure rats based on UPLC-TSQ-MS/MS[J]. Journal of Ethnopharmacol,2023,317:116818. doi: 10.1016/j.jep.2023.116818 -
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