Departamento de Biotecnología, Universidad Autónoma Metropolitana-Iztapalapa (UAM-I), Av. San Rafael Atlixco 186, Colonia Vicentina, Delegación Iztapalapa, 09340, Ciudad de México (CDMX), Mexico.
Centro de Investigación en Recursos Bióticos, Facultad de Ciencias, Universidad Autónoma del Estado de México (UAEMex), Carretera Toluca-Ixtlahuaca Km 14.5, Colonia San Cayetano, 50295, Toluca, Estado de México, Mexico.
J Ethnopharmacol. 2019 Aug 10;240:111875. doi: 10.1016/j.jep.2019.111875. Epub 2019 Apr 26.
Buddleja cordata Humb. Bonpl. & Kunth, known by the population as Tepozán blanco, is a shrub plant used in traditional herbal medicine in Mexico for the treatment of tumors, cancer, sores, skin burns, rheumatic pains and diseases related to inflammatory processes such as arthritis; authors adjudicate this etno-medicinal effect to the presence of secondary metabolites in the plant such as verbascoside, however due to its low concentration in recent years biotechnological tools are applied as cell culture to biosynthesize these pharmacological active metabolites in greater quantities.
Evaluate the possible toxic effect after a daily administration of MeOH extracts from wild plant leaves (Bc-Wp), and cell culture (Bc-Cc) of B. cordata for 28 days, and after their anti-edematous and antioxidant activities in vivo, as well their effect on the cytokines profile during experimental arthritis induced by complete Freund's adjuvant.
Both extracts were evaluated in CD1 male mice first in a toxicity test of repeated dose administrations (1 g/kg) for 28 days, after which pharmacological activity of both extracts was measure during experimental induced arthritis where three doses were tested, at the end of the study edema formation, body weight gain and antioxidant activity were measure in edema and ganglionic tissues. Finally, dose that exerted the best protective effect (250 mg/kg) was evaluated to quantify its effect over ganglionic tissue concentration of lymphocytes T CD4, and cytokines (IL-1β, TNF-α and IL-10), as well histological analysis in arthritic mice.
Both extracts (Bc-Wp and Bc-Cc) did not exert lethality, however body weight gain and food in-take were lower than in healthy mice administered with vehicles, also extract-treated animals showed a decrease in serum lipid concentration and only Bc-Wp extract treated animals decrease serum alkaline phosphatase after 28 daily administration compared to healthy un-treated mice. During experimental arthritis best inhibition effect over edema development was observed in those animals administered with both extracts at dose of 250 mg/kg (Bc-Wp and Bc-Cc) on day 28, compared to CFA un-treated mice. Also both extracts reduce oxidative damage over lipids and proteins at the same dose, in both ganglionic and edema tissue, as well antioxidant enzymatic response was reduced in both tissues compared to arthritic un-treated group. In ganglionic tissue of arthritic mice, CD4 lymphocytes concentration was reduced by Bc-Wp and Bc-Cc treatment (250 mg/kg) respectively, as well IL-1β, and TNF-α levels. Only arthritic animals treated with Bc-Cc extract at 250 mg/kg generated a significant increase of IL-10 doubling the levels compared to CFA un-treated group. Histological analysis of popliteal ganglion showed that both extracts decrease the incidence of lytic lesions, lipid inclusions and leukocyte infiltration.
Methanolic extracts of wild Buddleja cordata and its cell cultures did not generated lethality after a daily administration for 28 days at 1 g/kg, but it was observed that both showed a lipid-lowering effect. Also at dose of 250 mg/kg both extracts exerted anti-edematous, protection against the oxidation of lipid and proteins, regulation on antioxidant enzymatic response, down-regulation on lymphocytes CD4 producers of IL-1β and TNF-α, an increase in IL-10 levels, which caused a decrease in leukocyte infiltration in ganglionic tissue during experimental arthritis.
Buddleja cordata Humb. Bonpl. & Kunth，俗称 Tepozán blanco，是一种灌木植物，在墨西哥传统草药医学中用于治疗肿瘤、癌症、溃疡、皮肤烧伤、风湿痛和与炎症过程相关的疾病，如关节炎；作者认为这种民间医学作用归因于植物中存在的次生代谢物，如毛蕊花糖苷，然而，由于其近年来浓度较低，生物技术工具已被应用于细胞培养，以生物合成这些具有药理活性的代谢物。
评估每日给予野生植物叶片（Bc-Wp）和细胞培养物（Bc-Cc）的甲醇提取物 28 天后，以及体内抗水肿和抗氧化活性后，B. cordata 可能产生的毒性作用，以及其对完全弗氏佐剂诱导的实验性关节炎中细胞因子谱的影响。
首先，在重复剂量给药（1g/kg）28 天的毒性试验中，对两种提取物进行了评估，然后在实验性关节炎中测量了两种提取物的药理学活性，在研究结束时，测量了水肿形成、体重增加和抗氧化活性在水肿和神经节组织中。最后，评估了发挥最佳保护作用的剂量（250mg/kg），以定量评估其对神经节组织中 T 淋巴细胞 CD4 和细胞因子（IL-1β、TNF-α和 IL-10）浓度的影响，以及关节炎小鼠的组织学分析。
两种提取物（Bc-Wp 和 Bc-Cc）均未产生致死性，但与给予载体的健康小鼠相比，体重增加和食物摄入减少，接受提取物治疗的动物血清脂质浓度降低，仅 Bc-Wp 提取物治疗的动物在 28 天的每日给药后血清碱性磷酸酶降低，与未接受健康治疗的小鼠相比。在实验性关节炎中，与未接受 CFA 治疗的关节炎小鼠相比，在第 28 天给予 250mg/kg 剂量的两种提取物（Bc-Wp 和 Bc-Cc）时，对水肿发展的抑制作用最佳。两种提取物还可降低相同剂量的神经节和水肿组织中脂质和蛋白质的氧化损伤，以及两种组织中抗氧化酶反应的降低。在关节炎小鼠的神经节组织中，Bc-Wp 和 Bc-Cc 处理（250mg/kg）分别降低了 CD4 淋巴细胞的浓度，以及 IL-1β和 TNF-α水平。只有接受 Bc-Cc 提取物（250mg/kg）治疗的关节炎动物产生了 IL-10 的显著增加，与未接受 CFA 治疗的关节炎组相比，IL-10 的水平增加了一倍。对腘神经节的组织学分析表明，两种提取物均可减少溶骨性病变、脂质包涵体和白细胞浸润的发生率。
28 天内每天给予 1g/kg 的野生 Buddleja cordata 及其细胞培养物的甲醇提取物没有产生致死性，但观察到两者均有降低血脂的作用。此外，在 250mg/kg 剂量下，两种提取物均具有抗水肿、防止脂质和蛋白质氧化、调节抗氧化酶反应、下调产生 IL-1β和 TNF-α的 CD4 淋巴细胞、增加 IL-10 水平的作用，从而减少关节炎时神经节组织中的白细胞浸润。
