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1.南京中医药大学附属医院骨伤科,南京 210029
2.江苏省中医院骨伤科,南京 210029
硕士研究生。研究方向:中药防治膝骨关节炎。 E-mail:1184518489@qq.com
副主任药师,博士。研究方向:中医药防治膝骨关节炎。E-mail:liuzixiu3221@126.com
纸质出版日期:2023-12-15,
收稿日期:2023-06-12,
修回日期:2023-10-07,
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冯明庆,杨楠,方媛等.三色散凝胶贴膏原料药提取工艺的优化 Δ[J].中国药房,2023,34(23):2841-2847.
FENG Mingqing,YANG Nan,FANG Yuan,et al.Optimization of the extraction technology for the raw drugs of Sanse powder gel paste[J].ZHONGGUO YAOFANG,2023,34(23):2841-2847.
冯明庆,杨楠,方媛等.三色散凝胶贴膏原料药提取工艺的优化 Δ[J].中国药房,2023,34(23):2841-2847. DOI: 10.6039/j.issn.1001-0408.2023.23.05.
FENG Mingqing,YANG Nan,FANG Yuan,et al.Optimization of the extraction technology for the raw drugs of Sanse powder gel paste[J].ZHONGGUO YAOFANG,2023,34(23):2841-2847. DOI: 10.6039/j.issn.1001-0408.2023.23.05.
目的
2
对三色散凝胶贴膏原料药的提取工艺进行优化。
方法
2
将SD大鼠分为空白组、模型组、传统工艺组、水提组和醇提组,每组5只,采用前交叉韧带横断术建立膝骨关节炎模型,考察不同提取方式所得凝胶贴膏对膝骨关节炎大鼠的药效学影响。分别以冷、热痛阈值与疼痛介质降钙素基因相关肽(CGRP)、环氧合酶2(COX-2)、P物质(SP)、前列腺素E2(PGE2)含量为指标进行镇痛实验;对大鼠滑膜组织进行HE染色,观察大鼠滑膜细胞增殖、炎症细胞浸润、血管入侵程度,并以白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、IL-6蛋白与mRNA表达水平为指标进行抗炎实验,对比各组大鼠的抗炎镇痛效果差异。以加醇倍数、提取时间、提取次数作为正交试验考察因素,以蔓荆子黄素、士的宁和马钱子碱含量为评价指标并计算综合评分,优选出三色散凝胶贴膏原料药的提取工艺并进行验证实验。
结果
2
与模型组比较,各给药组大鼠的冷痛阈值和热痛阈值(除传统工艺组外)均显著升高(
P
<0.05),疼痛介质CGRP、COX-2、SP、PGE2含量均显著下降(
P
<0.05);滑膜组织炎症细胞浸润减少、纤维化程度减轻,胶原沉积减少,可见少量毛细血管增生;滑膜组织中IL-1β、IL-6、TNF-α蛋白与mRNA表达水平均显著降低(
P
<0.05)。与传统工艺组比较,醇提组大鼠的大部分检测指标显著降低(
P
<0.05),而水提组大鼠仅热痛阈值和滑膜组织中IL-6 mRNA表达水平显著降低(
P
<0.05)。三色散凝胶贴膏原料药的最佳提取工艺为取适量三色散细粉,加8倍量55%乙醇,加热回流提取90 min,提取2次;3次验证实验结果显示,蔓荆子黄素、士的宁与马钱子碱含量平均值分别为0.007%、0.092%、0.214%,RSD均小于5%。
结论
2
优化的三色散凝胶贴膏原料药提取工艺稳定、可行,有利于提高该制剂药效。
OBJECTIVE
2
To optimize the extraction technology for the raw drugs of Sanse powder gel paste.
METHODS
2
SD rats were divided into blank group, model group, traditional technology group, water extraction group and ethanol extraction group, with 5 rats in each group. Anterior cruciate ligament transection was used to construct knee osteoarthritis model, and the pharmacodynamic effects of different extraction methods on arthritic rats were investigated. Analgesic experiments were conducted using cold and hot pain thresholds and pain mediators calcitonin gene-related peptide (CGRP), cyclooxygenase-2 (COX-2), substance P (SP), and prostaglandin E2 (PGE2) contents as indicators. HE staining was performed on the synovial membrane of rats to observe the degree of synovial cell proliferation, inflammatory infiltration and vascular invasion, and anti-inflammatory experiments were conducted using protein and mRNA expressions of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and IL-6 as indicators. The analgesic and anti-inflammatory effects were compared among those groups. In the orthogonal test, ethanol dosage, extraction time and extraction times were used as evaluation factors, and the contents of casticin, strychnine and toxiferine were taken as evaluation indicators; comprehensive score was calculated. The validation experiments were carried out after optimizing the extraction technology of the raw drugs of Sanse powder gel paste.
RESULTS
2
Compared with the model group, the cold and heat pain thresholds of drug administration groups (except for the traditional technology group) were all increased significantly (
P
<0.05), while the contents of pain mediators CGRP, COX-2, SP and PGE2 were all decreased significantly (
P
<0.05). HE staining showed that inflammatory cell infiltration, fibrosis and collagen deposition were decreased in the administration groups; a small amount of capillary proliferation could be found; the protein and mRNA expressions of inflammatory factors such as IL-1β, IL-6 and TNF-α were decreased significantly in synovial tissue of rats in administration groups (
P
<0.05). Compared with the traditional technology group, most indicators of the ethanol extraction group were significantly reduced (
P
<0.05), and only heat pain threshold and mRNA expression of IL-6 in rats were decreased significantly in the water extraction group (
P
<0.05). The optimal extraction technology of the raw drugs of Sanse powder gel paste included suitable dose of Sanse powder, 8-fold 55% ethanol, heating reflux extraction for 90 minutes, extracting twice. The results of 3 times of verification experiments showed that the average contents of casticin, strychnine and toxiferine were 0.007%, 0.092%, and 0.214%, respectively; RSD were all less than 5%.
CONCLUSIONS
2
The optimized extraction technology for the raw drugs of Sanse powder gel paste is stable and feasible, which can improve the efficacy of the preparation.
三色散凝胶贴膏提取工艺药效学含量测定
gel pasteextraction technologypharmacokineticscontent determination
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