基于血浆NO/IL-4水平研究葛根芩连汤整方不同剂量治疗溃疡性结肠炎的量-效关系

Dose-response relationship based on plasma NO/IL-4 levels for different doses of Gegen Qinlian Decoction in the treatment of ulcerative colitis

摘要

目的 基于血浆NO/IL-4水平探讨葛根芩连汤(GQD)整方不同剂量治疗溃疡性结肠炎(UC)的量-效关系,并计算相关剂量参数,为临床用药提供理论剂量参考。方法 120只雄性SD大鼠,除正常组外,其余各组以50 g/L的2,4,6-三硝基苯磺酸(TNBS)+500 mL/L乙醇灌肠诱导大鼠UC模型,造模大鼠分为模型组、柳氮磺吡啶组及GQD的9个剂量组(1.65、4.95、8.25、11.55、14.85、18.15、21.45、24.75、28.05 g/kg),每组8只,给药1次/d,共5 d。观察大鼠一般情况及结肠组织损伤的病理变化,计算结肠黏膜损伤指数,检测大鼠血浆白介素-4(IL-4)、一氧化氮(NO)水平并经GraphPad Prism 9绘制、拟合量效曲线,计算相关剂量参数。结果 与模型组比较,GQD 18.15 g/kg、28.05 g/kg组可显著降低大鼠结肠溃疡评分(P＜0.01)和结肠黏膜损伤指数(P＜0.05);GQD(1.65、4.95、8.25、11.55、14.85、18.15 g/kg)组的大鼠血浆NO水平显著降低(P＜0.05);GQD(1.65、4.95、8.25、11.55、14.85 g/kg)组的大鼠血浆IL-4水平显著增高(P＜0.01)。分别以NO、IL-4数据为指标,计算得ED₅₀=16.08 g/kg,ED₂₀～ED₈₀=13.51～19.14 g/kg;ED₅₀=15.71 g/kg,ED₂₀～ED₈₀=14.86～16.60 g/kg。结论 GQD可能通过降低NO和升高IL-4水平改善UC,且存在一定的量效关系,其治疗范围大致为14.86～16.60 g/kg。

Abstract

Objective To explore the dose-response relationship of different doses of the whole formula of Gegen Qinlian Decoction (GQD) in the treatment of ulcerative colitis (UC) based on the levels of plasma nitric oxide (NO) and interleukin-4 (IL-4), and to calculate the relevant dose parameters, so as to provide theoretical dose reference for clinical medication. Methods A total of 120 male SD rats were selected. Except for the normal group, the rats in the other groups were induced to establish UC models by enema with 50 g/L 2,4,6-trinitrobenzene sulfonic acid (TNBS) + 500 mL/L ethanol. After successful modeling, the rats were divided into model group, sulfasalazine group and 9 GQD dose groups (1.65, 4.95, 8.25, 11.55, 14.85, 18.15, 21.45, 24.75, and 28.05 g/kg), with 8 rats in each group. The drugs were administered once a day for five consecutive days. The general condition of the rats and the pathological changes of colonic tissue damage were observed; the colonic mucosal damage index was calculated. The levels of plasma IL-4 and NO in rats were detected, and the dose-effect curve was drawn and fitted by GraphPad Prism 9 software to calculate the relevant dose parameters. Results Compared with the model group, the GQD 18.15 g/kg and 28.05 g/kg groups had significantly reduced colonic ulcer score (P＜0.01) and colonic mucosal damage index (P＜0.05) of rats; the plasma NO level of rats in the GQD (1.65, 4.95, 8.25, 11.55, 14.85, and 18.15 g/kg) groups was extremely significantly reduced (P＜0.05); the plasma IL-4 of rats in the GQD (1.65, 4.95, 8.25, 11.55, and 14.85 g/kg) groups was significantly increased (P＜0.01). Taking the plasma levels of NO and IL-4 as detection indicators, the dose parameters were calculated respectively: ED₅₀=16.08 g/kg, ED₂₀-ED₈₀=13.51-19.14 g/kg; ED₅₀=15.71 g/kg, ED₂₀-ED₈₀=14.86-16.60 g/kg. Conclusion GQD may improve UC by reducing plasma NO level and increasing IL-4 level, and there is an obvious dose-response relationship. Its recommended clinical treatment range is roughly 14.86-16.60 g/kg.

关键词

葛根芩连汤 / 溃疡性结肠炎 / 量-效关系 / 炎症因子 / 白介素-4(IL-4) / 一氧化氮(NO)

Key words

Gegen Qinlian Decoction / ulcerative colitis / dose-response relationship / inflammatory factor / interleukin-4 (IL-4) / nitric oxide (NO)

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基金资助

江门市医疗卫生领域科技计划项目(2024YL05001)

广东江门中医药职业学院科技计划项目(JMZYYKY20251003)