有毛白花苋提取物对乙二醇诱导的雄性白化Wistar大鼠肾结石的防治作用

Preventive and therapeutic effects of Aerva lanata(L.) extract on ethylene glycol-induced nephrolithiasis in male Wistar albino rats

目的 肾结石是一种常见的泌尿外科疾病,本研究主要评估了有毛白花苋根的水醇提取物(HAEAL)对乙二醇诱导的大鼠肾结石的防治作用。方法 使用索氏提取器,将50 g阴干的有毛白花苋根粗研粉末,按照有机溶剂极性递增的顺序,依次通过石油醚(60-80℃)、氯仿和乙醇提取后浓缩。同时,对草药悬浮液的性质、颜色、气味和质地进行物理检测。肾结石体外评估通过成核试验、聚集试验和晶体生长试验进行。将30只成年雄性Wistar白化大鼠随机分为5组,每组6只。第一组为阴性对照组,不进行结石诱导并自由饮水28天;第二组为阳性对照组,口服0.75%乙二醇处理14天,并自由饮水28天;第三组为标准组,口服0.75%乙二醇处理14天,第15天至第28天予以750 mg/kg Cystone;第四组为低剂量HAEAL组,口服0.75%乙二醇处理14天,第15天至第28天给予400 mg/kg HAEAL,每天1mL;第五组为高剂量HAEAL组,口服0.75%乙二醇处理14天,第15天至第28天给予800 mg/kg HAEAL,每天1mL;之后进行尿液(尿量、pH值、外观、气味及浑浊度)检验和血清试验。第29天,对大鼠肾脏进行解剖,通过组织病理学检查,以确定肾实质的损伤程度。结果在室温下,悬浮液具有芳香气味,且稳定性良好,无浑浊现象。至第三天,悬浮液未出现颜色及气味的变化,说明该制剂的稳定性可达72小时。阳性对照组的大鼠体重减轻,表明有结石形成及饮水量改变。标准治疗组和HAEAL治疗组在接受治疗后,体重均恢复到正常水平,表明治疗效果良好。组织病理学检查结果显示,阴性对照组无明显病变,阳性对照组大鼠的肾实质区域出现炎症反应。与阳性对照组相比,HAEAL治疗组大鼠的尿量和尿成分如钙和草酸的排泄均有增加(P <0.01),且清除率也有所提高(P <0.05)。此外,HAEAL治疗组大鼠的尿液pH值显著升高。结论 HAEAL能减少肾结石形成,且具有利尿作用,可促进结石的排出。但对于如何提高悬浮液的稳定性,以制备更好的药物配方,仍需进一步的研究。

Objective Nephrolithiasis is a common urological disease. This study aims to evaluate the preventive and therapeutic effects of hydro-alcoholic extract of Aerva lanata(L.) roots(HAEAL) on ethylene glycol-induced nephrolithiasis in rats.Methods Fifty grams of shade-dried coarsely powdered Aerva lanata(L.) root was successively extracted with organic solvents in increasing order of polarity [petroleum ether(60-80 ℃), chloroform, and ethanol] using a Soxhlet apparatus, and then concentrated. Physical tests including nature, color, odor, and texture were performed on the herbal suspension. In vitro nephrolithiasis assessment was performed by nucleation assay, aggregation assay, and crystal growth assay. Thirty adult male Wistar albino rats were randomly divided into five groups(six rats in each group). Group 1: negative control group without induction or treatment till day 28. Group 2: positive control group receiving a daily oral solution of 0.75% ethylene glycol till day 14, and mixed with distilled water till day 28. Group 3: standard group receiving a daily oral solution of 0.75% ethylene glycol till day 14 and Cystone(750 mg/kg) from day 15 to day 28. Group 4: low dose HAEAL group receiving a daily oral solution of 0.75%ethylene glycol till day 14, and 400 mg/kg HAEAL from day 15 to day 28(1 mL per day). Group 5: high dose HAEAL group receiving a daily oral solution of 0.75% ethylene glycol till day 14,and 800 mg/kg HAEAL from day 15 to day 28(1 mL per day). Urine(urine volume, pH value,appearance, odor, and turbidity) examination and serum test were performed. On day 29, the kidneys were dissected, and histopathology examination was performed to determine the degree of tubular injury.Results The suspension showed stability and aroma with no turbidity at room temperature.The suspension did not show changes in color and odor until day 3, indicating that the preparation was stable for 72 h. Body weight decreased in the positive control group indicating stone formation and changes in water intake. Both standard and HAEAL treatments restored the body weight to normal levels after treatment, indicating the beneficial effects of the treatment. Histopathological examination revealed no significant findings in the negative control group, whereas the positive control group showed inflammation in the kidney parenchyma.Compared with positive control group, there was increase in urine volume and excretion of urinary constituents such as calcium and oxalate(P < 0.01) as well as improved clearance rate(P < 0.05) in HAEAL treatment groups, in addition, the urine pH value of HAEAL groups was increased.Conclusion HAEAL reduced nephrolithiasis formation and had a diuretic effect, which could be used to promote the expulsion of stones. Further studies are needed to enhance the stability of the suspension for the production of better pharmaceutical formulations.