雷公藤甲素对糖尿病模型大鼠的经皮渗透特性研究

Study on the Permeability of Triptolide in the Skin of Model Rats with Diabetes

目的:研究雷公藤甲素经糖尿病模型大鼠皮肤的渗透特性。方法:一次性ip给予链脲佐菌素(55 mg/kg)以复制大鼠糖尿病模型,剥离鼠皮备用,同时设正常对照(正常大鼠皮)组与模型组。采用改良型Franz扩散池评价雷公藤甲素经皮渗透效果并测定大鼠皮肤各层中雷公藤甲素滞留量,采用高效液相色谱串联质谱(LC-MS/MS)法检测雷公藤甲素含量。色谱柱为Agilent ZORBAX-C18,流动相为甲醇-水(60∶40),流速为0.3 ml/min,进样量为2μl,柱温为30℃;电喷雾电离(ESI)+,电离电压为+4.0 k V,检测离子对为m/z 361.1→91.1,离子源温度为110℃,干燥气温度为350℃,流量为7 L/min,监测模式为多重反应检测(MRM)模式,裂解电压为135 V,碰撞能量为55 e V。结果:模型组大鼠皮的稳态渗透速率为12.66μg/(h·cm2),单位面积累积透过量为(165.42±8.90)μg/cm2;与正常对照组(19.22±4.32)μg/cm2比较,上述两个指标均明显增加(P<0.05)。模型组大鼠真皮层中药物含量为(34.32±0.47)μg/cm2,明显高于正常对照组大鼠真皮层的(12.48±0.35)μg/cm2。结论:糖尿病模型大鼠皮肤对雷公藤甲素的渗透性(渗透速率、单位面积累积透过量、皮肤滞留量)明显增加。

OBJECTIVE： To study the permeability of triptolide in the skin of model rats with diabetes. METHODS： Strepto- zotocin （55 mg/kg） was given at a time to establish rat models of diabetes, ip. The skins of the rats were peeled off. There was a normal control group （normal rat skins） and model group. The modified franz diffusion cell was employed to evaluate the permea- bility of permeability of triptolide in the skin, and the retention amounts of permeability of triptolide in the layers of rats＇ skins were determined. Liquid chromatography-mass spectrometry （LC-MS/MS） was adopted to determine the content of permeability of triptolide. The determination was performed on Agilent ZORBAX-C18 with the mobile phase of methanol-water （60 ： 40）, at the flow rate of 0.3 ml/min, with the sample volume of 2 μl, the column temperature of 30℃. Electrospray ionization （ESI）＋ was per- formed at the ionization voltage of ＋4.0 kV, with the ion pair of m/z 361.1→91.1 being tested, ion source temperature of 110℃, dry gas temperature of 350 ℃, flow of 7 L/min, multiple reaction monitoring （MRM） as the monitoring mode, fragmentor voltage of 135 V and collision energy of 55 eV. RESULTS： The steady state permeation rate in model group was 12.66 pg/（h·cm^2） and the cumulative permeation amount was （165.42 ± 8.90） μg/cm^2; compared with normal control group, the above-mentioned indexes were obviously increased （P〈0.05）. The content of the drug in the corium layer of the rats in model group was （34.32 ± 0.47）μg/cm^2, which was significantly higher than （12.48 v 0.35） μg/cm^2 of normal rats. CONCLUSIONS： The skin permeability （perme- ation rate, cumulative permeation amount and the retention amount in the skin） to the triptolide for the rats with diabetes is obvious- ly increased.