[关键词]
[摘要]
目的 研究纹党Codonopsis Radix米炒前后体外抗氧化活性变化规律及干预脾虚泄泻大鼠的药效差异。方法 采用1,1-二苯基-2-三硝基苯肼(DPPH)和2,2-氮杂双-(3-乙基苯并噻唑啉-6-磺酸)二铵盐(ABTS)2种方法,考察纹党米炒前后体外抗氧化活性的变化规律。建立脾虚泄泻大鼠模型,给予硫酸阿托品注射液、纹党和米炒纹党水提液进行干预,记录各组大鼠体质量;计算各组大鼠脾脏指数和胸腺指数;采用ELISA法检测各组大鼠血清中胃动素(motilin,MTL)、胃泌素(gastrin,GAS)、血管活性肠肽(vasoactive intestinal peptide,VIP)、淀粉酶(amylase,AMS)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)和IL-10水平;采用苏木素-伊红(HE)染色法观察各组大鼠十二指肠、空肠组织病理改变情况,比较纹党米炒前后治疗脾虚泄泻大鼠在免疫调节和消化吸收方面的疗效差异。结果 体外抗氧化结果显示,纹党、米炒纹党的清除DPPH自由基半数抑制浓度(half inhibitory concentration,IC50)分别为(0.041±0.110)、(0.029±0.140)g/mL;其清除ABTS自由基IC50分别为(0.038±0.050)、(0.029±0.100)g/mL。药效结果显示,与对照组比较,模型组大鼠体质量、胸腺指数、脾脏指数、TNF-α、IL-10、MTL、GAS、VIP及AMS水平明显下降(P<0.05),IL-6水平明显上升(P<0.05),小肠病理切片结果显示十二指肠、空肠病变严重。与模型组比较,各给药组脾脏指数及血清TNF-α、IL-10、IL-6水平均具有明显差异(P<0.05);除纹党低剂量组胸腺指数,纹党中、低剂量组与米炒纹党低剂量组MTL水平以及纹党低剂量组GAS水平在统计学上没有差异外,其余各给药组均可使大鼠胸腺指数、MTL、GAS、VIP及AMS水平明显上升(P<0.05),各给药组十二指肠、空肠病变不同程度地恢复。与相同剂量生品组比较,米炒纹党高、中剂量组脾脏指数及米炒纹党高、低剂量组胸腺指数明显上升(P<0.05),米炒纹党高、中、低剂量组TNF-α、IL-10、GAS及VIP水平,米炒纹党高剂量组MTL水平以及米炒纹党中剂量组AMS水平明显上升(P<0.05),米炒纹党高、中剂量组IL-6水平明显下降(P<0.05)。结论 纹党米炒后抗氧化活性增强;纹党生熟饮片对脾虚泄泻大鼠免疫调节和消化吸收方面均具有明显治疗作用,且米炒品明显强于生品,可能与调节TNF-α、IL-6、IL-10水平密切相关,说明米炒纹党增强和胃健脾止泻的科学性、合理性。
[Key word]
[Abstract]
Objective To comparative the effects of Wendang (Codonopsis Radix) before and after stir-baking with rice on the antioxidation in vitro and regulation of immune function and digestive absorption in diarrhea rats with spleen deficiency. Methods DPPH and ABTS methods were used to investigate the changes of antioxidant activity of Codonopsis Radix before and after stir-baking with rice. The model of diarrhea of rats with spleen deficiency was established. The sulfate injection and water extract of Codonopsis Radix before and after stir-baking with rice were given for intervention, body weight of rats in each group was recorded, spleen index and thymus index of rats in each group were calculated. Levels of MTL, GAS, VIP, AMS, TNF-α, IL-6, and IL-10 in serum of each group of rats were determined by ELISA. HE staining method was used to observed the pathological changes of duodenum and jejunum of each group of rats. The effects of Codonopsis Radix before and after stir-baking with rice on diarrhea of spleen deficiency rats in terms of immune regulation, digestion and absorption were compared. Results Results of antioxidation in vitro showed that IC50 of scavenging DPPH free radical of Codonopsis Radix before and after stir-baking with rice were (0.041 ±0.110) and (0.029 ±0.140) g/mL, IC50 of scavenging ABTS free radical were (0.038 ±0.050) and (0.029 ±0.100) g/mL. Compared with control group, body weight, thymus index, spleen index, TNF-α, IL-10, MTL, GAS, VIP and AMS levels in model group were significantly decreased (P<0.05), while the level of IL-6 was significantly increased (P<0.05). The pathological changes of duodenum and jejunum were severe in model group. The treatment of each experimental group had different effects on spleen index, TNF-α, IL-10 and IL-6 as compared to model group. There was no statistical difference in thymus index between low-dose Codonopsis Radix group, the level of MTL was not statistically different between medium- and low-dose Codonopsis Radix groups and low-dose Codonopsis Radix after stir-baking with rice group, and level of GAS in low-dose Codonopsis Radix group had no statistically difference. All the other groups could significantly increase the thymus index, MTL, GAS, VIP and AMS levels in rats (P<0.05). The pathological changes of duodenum and jejunum were ameliorated in different degrees in each treatment group. Compared with same dose group, spleen index of high- and medium-dose Codonopsis Radix after stir-baking with rice group and thymus index of high- and low-dose Codonopsis Radix after stir-baking with rice group were significantly increased (P<0.05). The levels of TNF-α, IL-10, GAS and VIP in high-, medium- and low-dose Codonopsis Radix after stir-baking with rice groups, MTL level in high-dose Codonopsis Radix after stir-baking with rice group, and AMS levels in medium- dose Codonopsis Radix after stir-baking with rice group were significantly increased (P<0.05), the level of IL-6 in high- and medium-dose Codonopsis Radix after stir-baking groups with rice was significantly decreased (P<0.05). Conclusion Antioxidant activity of Codonopsis Radix was enhanced after stir-baking with rice. Raw and cooked slices of Codonopsis Radix have obvious therapeutic effects on diarrhea in rats with spleen deficiency in therms of immune regulation, digestion and absorption. Codonopsis Radix after stir-baking with rice is obviously stronger than raw products, which may be closely related to the regulation of levels of TNF- α, IL-6 and IL-10, which demonstrates that Codonopsis Radix after stir-baking with rice can harmonize stomach, invigorate and relieve diarrhea scientifically and reasonably.
[中图分类号]
R285.5
[基金项目]
国家重点研发计划(2018YFC1706303);全国中药特色技术传承人才培训项目(T20184828005)