数据挖掘的中药新方对卵巢储备功能下降大鼠干预的实验分析

Intervention Effect of New Chinese Herbal Prescription Selected by Data Mining Against Decreased Ovarian Reserve in Rats

目的:遴选及评估用于治疗卵巢储备功能下降(DOR)的中药新方及其适宜剂量。方法:检索中国期刊全文数据库、万方、维普等数据库,收集有关中医药治疗DOR的文献,应用中医传承辅助平台V2.5建立数据库并进行数据挖掘得出药物核心组合及中药新方组合,采用专家小组讨论法确定用于实验评估的中药新方。将SD雌性大鼠分为空白组,DOR模型组,坤泰胶囊组,中药新方(简称新方)低、中、高剂量组,每组12只大鼠。坤泰胶囊组,新方低、中、高剂量组分别给DOR大鼠灌胃坤泰胶囊溶液(按说明书用量折算为0.5 g·kg^(-1))和3.0375,6.075,9.12 g·kg^(-1)新方水煎液进行干预治疗。21 d后,阴道脱落细胞涂片观察大鼠动情周期,苏木素-伊红(HE)染色观察卵巢组织结构,生化法检测血清中抗穆勒氏管激素(AMH),卵泡刺激素(FSH),黄体生成素(LH),雌二醇(E;)含量和卵巢中活性氧(ROS),丙二醛(MDA),谷胱甘肽(GSH)含量及超氧化物歧化酶(SOD),过氧化氢酶(CAT),谷胱甘肽过氧化物酶(GSH-Px)活性。结果:用于实验评估的中药新方由熟地黄20 g,鹿角胶12 g,枸杞子20 g,山茱萸12 g,合欢皮9 g,莲子心3 g,党参20 g,黄芪30 g,白术12 g,山药30 g,炙甘草6 g共11味药组成;与模型组相比,坤泰胶囊组、新方中、高剂量组动情周期、卵泡发育明显改善,血清中AMH,E;含量明显升高(P<0.05),FSH和LH含量明显降低(P<0.05),卵巢中ROS和MDA含量明显降低(P<0.05),SOD,CAT,GSH-Px活力及GSH含量明显升高(P<0.05)。与坤泰胶囊组相比,新方中、高剂量组的上述指标没有显著差异,而动情周期和卵泡发育呈现更好。结论:通过数据挖掘筛选的中药新方可提高DOR大鼠卵巢抗氧化,促进卵泡发育,改善血清激素水平和动情周期,有效提高卵巢储备功能,最适浓度可选择为中剂量6.075 g·kg^(-1)。

Objective:To select and evaluate new Chinese herbal prescription for the treatment of decreased ovarian reserve(DOR)and its appropriate dosage.Method:The literature concerning the treatment of DOR with traditional Chinese medicine(TCM)was retrieved from such databases as Chinese Journal Fulltext Database,Wanfang Data Knowledge Service Platform,and Chongqing Weipu Database for Chinese Technical Periodicals(VIP),based on which a database was established using the Traditional Chinese Medicine Inheritance Support System(TCMISS)V2.5.The data mining was then carried out to obtain the core combinations of Chinese herbs and new Chinese herbal prescription combinations,followed by the determination of the new Chinese herbal prescription by expert group discussion for experiment evaluation.The female SD rats were divided into the normal group,DOR model group,Kuntai capsule group,and low-,medium-,and highdose new Chinese herbal prescription groups,with 12 rats in each group.Rats in the Kuntai capsule group and low-,medium-,and high-dose new Chinese herbal prescription groups were treated with Kuntai capsule solution(0.5 g·kg^(-1)determined according to the dosage in the instruction)and 3.0375,6.075,and 9.12 g·kg^(-1)new Chinese herbal prescription,respectively.After 21 days,the estrous cycle was observed by vaginal exfoliated cell smear,and the ovarian structure was observed by hematoxylin-eosin(HE)staining.The serum antimullerian hormone(AMH),follicle stimulating hormone(FSH),luteinizing hormone(LH),and estrogen(E;)contents as well as the reactive oxygen species(ROS),malondialdehyde(MDA),and glutathione(GSH)levels and superoxide dismutase(SOD),catalase(CAT),and glutathione peroxidase(GSH-Px)activities in ovary were detected using biochemical methods.Result:The new Chinese herbal prescription subjected to experimental evaluation was composed of 11 Chinese herbs,namely Rehmanniae Radix Praeparata 20 g,Cervi Cornus Colla 12 g,Lycii Fructus 20 g,Corni Fructus 12 g,Albiziae Cortex 9 g,Nelumbinis Plumula 3 g,Salviae Miltiorrhizae Radix et Rhizoma 20 g,Astragali Radix 30 g,Atractylodis Macrocephalae Rhizoma 12 g,Dioscoreae Rhizoma 30 g,and Glycyrrhizae Radix et Rhizoma Praeparata Cum Melle 6 g.Compared with the model group,the Kuntai capsule group and medium-and high-dose new Chinese herbal prescription groups exhibited significantly improved estrous cycle and follicular development,elevated serum AMH and E;and ovarian GSH(P<0.05),decreased serum FSH and LH(P<0.05)and ovarian ROS and MDA(P<0.05),and enhanced SOD,CAT,and GSH-Px activities(P<0.05).There were no significant differences in the abovementioned indexes between the Kuntai capsule group and the middle-and high-dose new Chinese herbal prescription groups,but the estrous cycle and follicular development were better in the latter two groups.Conclusion:The new Chinese herbal prescription screened by data mining is able to enhance ovarian antioxidation,promote follicular development,ameliorate serum hormone and estrous cycle,and effectively improve ovarian reserve function in DOR rats.The medium dose(6.075 g·kg^(-1))has been proved optimal.