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瓦尼桑黄粗多糖的细胞毒性、急性和亚急性毒性以及抗氧化活性(体外)评估

Assessment of cytotoxicity, acute, subacute toxicities and antioxidant activities (in vitro) of Sanghuangporus vaninii crude polysaccharide.

作者信息

Liu Jinze, Song Jinyue, Chen WeiJia, Sun Li, Zhao Yan, Zong Ying, He Zhongmei, Du Rui

机构信息

College of Chinese Medicinal Materials, Jilin Agricultural University, Changchun, 130118, Jilin, China.

College of Chinese Medicinal Materials, Jilin Agricultural University, Changchun, 130118, Jilin, China; China Jilin Provincial Engineering Research Center for Efficient Breeding and Product Development of Sika Deer of China, Changchun, Jilin, 130118, China.

出版信息

J Ethnopharmacol. 2024 Jan 30;319(Pt 3):117284. doi: 10.1016/j.jep.2023.117284. Epub 2023 Oct 14.

Abstract

ETHNOPHARMACOLOGY RELEVANCE

Sanghuangporus vaninii (S. vaninii), as a traditional large medicinal fungus, has a history of more than 2000 years in Chinese history and has been widely used to treat female diseases such as vaginal discharge, amenorrhea, and uterine bleeding, and recent pharmacological studies have also found that it has antioxidant, anti-inflammatory, and anti-tumor physiological activity, which has received more and more attention.

AIM OF THE STUDY

The objective was to evaluate cytotoxicity and the acute, subacute toxicity, and in vitro antioxidant activity of S. vaninii crude polysaccharide (SVP).

MATERIALS AND METHODS

The monosaccharide composition of SVP was determined by HPLC (high-performance liquid chromatography). The cytotoxicity of different concentrations of SVP on three types of cells (HT-22, Kupffer macrophages, HEK293) was assessed using CCk-8. The acute toxicity in vivo was evaluated for 14 days after the administration of SVP (2500,5000, or 10,000 mg/mL). For the evaluation of subacute toxicity, mice were daily treated for 28 days with SVP (2500,5000, or 10,000 mg/mL). In addition, DPPH, hydroxyl radical, and superoxide anion radical were used to evaluate the in vitro antioxidant activity of SVP.

RESULTS

SVP was not toxic in all three cell lines tested. In vitro antioxidant tests on the extracts showed that SVP possessed a strong antioxidant capacity in vitro. In the acute study, the no-observed-adverse-effect level (NOAEL) in male and female rats was 10,000 mg/kg body weight. There were also no deaths or severe toxicity associated with SVP in subacute studies. However, SVP treatment had a decreasing effect on body weight in mice of both sexes (2500, 5000, and 10000 mg/kg). At doses (5000 and 10,000 mg/kg), SVP had a reduced effect on food intake in both male and female mice. In addition, there were significant effects on organ coefficients of the liver, lung, and kidney. Hematological analysis showed significantly lower LYM (%) values in mice of both sexes, with significantly lower MCH (pg) values obtained in males (5000 mg/kg and 10000 mg/kg) and higher GRAN (%) values in females. In addition, the RDW-SD (fL) values were significantly lower in the male mice given the highest dose. Biochemical tests showed that there were no significant changes in ALT, AST, TP, and Cr levels after SVP treatment. In histopathological analysis, mild liver toxicity was observed in both female mice treated with 10,000 mg/kg SVP.

CONCLUSION

The extract of SVP showed a predominance of polysaccharide compounds, with non-toxic action in vivo. Our approach revealed SVP on the chemical composition and suggests a high margin of safety in the popular use of medicinal fungi. In conclusion, our results suggest that SVP is safe, and can be used as health care products and food.

摘要

民族药理学相关性

瓦尼桑黄作为一种传统大型药用真菌,在中国已有2000多年历史,被广泛用于治疗诸如白带、闭经和子宫出血等妇科疾病,且近期药理学研究还发现它具有抗氧化、抗炎和抗肿瘤生理活性,已受到越来越多关注。

研究目的

旨在评估瓦尼桑黄粗多糖(SVP)的细胞毒性、急性和亚急性毒性以及体外抗氧化活性。

材料与方法

采用高效液相色谱法(HPLC)测定SVP的单糖组成。使用CCk - 8评估不同浓度SVP对三种细胞(HT - 22、库普弗巨噬细胞、HEK293)的细胞毒性。给予SVP(2500、5000或10000 mg/mL)后,评估其体内急性毒性14天。为评估亚急性毒性,小鼠每日用SVP(2500、5000或10000 mg/mL)处理28天。此外,采用二苯基苦味酰基自由基(DPPH)、羟基自由基和超氧阴离子自由基评估SVP的体外抗氧化活性。

结果

SVP在所测试的所有三种细胞系中均无毒性。对提取物的体外抗氧化测试表明,SVP在体外具有较强的抗氧化能力。在急性研究中,雄性和雌性大鼠的未观察到有害作用水平(NOAEL)为10000 mg/kg体重。在亚急性研究中,也未发现与SVP相关的死亡或严重毒性。然而,SVP处理对两性小鼠体重均有减轻作用(2500、5000和10000 mg/kg)。在剂量为5000和10000 mg/kg时,SVP对雄性和雌性小鼠的食物摄入量均有降低作用。此外,对肝脏、肺和肾脏的器官系数有显著影响。血液学分析显示,两性小鼠的淋巴细胞(LYM)(%)值均显著降低,雄性小鼠(5000 mg/kg和10000 mg/kg)的平均红细胞血红蛋白含量(MCH)(pg)值显著降低,雌性小鼠的粒细胞(GRAN)(%)值升高。此外,给予最高剂量的雄性小鼠的红细胞分布宽度标准差(RDW - SD)(fL)值显著降低。生化测试表明,SVP处理后谷丙转氨酶(ALT)、谷草转氨酶(AST)、总蛋白(TP)和肌酐(Cr)水平无显著变化。在组织病理学分析中,给予10000 mg/kg SVP的雌性小鼠均观察到轻度肝脏毒性。

结论

SVP提取物显示多糖化合物占优势,在体内无毒性作用。我们的研究方法揭示了SVP的化学成分,并表明在药用真菌的广泛应用中具有较高的安全性。总之,我们的结果表明SVP是安全的,可作为保健品和食品使用。

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