二神丸()提取物对脾(脾)肾(肾)阳虚型腹泻大鼠模型的治疗作用。

Therapeutic Effect of Ershen Pill () Extract on Pi (Spleen)-Shen (Kidney) Yang Deficiency-Induced Diarrhea in Rat Model.

机构信息

College of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu, 610075, China.

College of Pharmacy, South Central University for Nationalities, Wuhan, 430074, China.

出版信息

Chin J Integr Med. 2019 Jul;25(7):536-542. doi: 10.1007/s11655-017-2766-4. Epub 2018 Mar 21.

Abstract

OBJECTIVE

To investigate whether Ershen Pill (ESP, ) could alleviate the symptom of Pi (Spleen)-Shen (Kidney) yang deficiency (PSYD)-induced diarrhea in rat model and explore its anti-diarrhea mechanism.

METHODS

Seventy-five Sprague-Dawley rats were divided into 5 groups by a random number table, including control, positive, model, low-dose (LD) and high-dose (HD) ESP groups, 15 rats in each group. All the rats, except those in the control group, were developed PSYD induced-diarrhea based on its pathology and etiology. The rats in positive, LD and HD ESP groups were treated with Shenling Baizhu Pill (), LD (1.05 g/kg) or HD (3.50 g/kg) ESP petroleum ether extract once a day for 2 weeks, respectively. Body weight change and diarrhea index were measured. The histology scores of the kidney were evaluated via hematoxylin and eosin (HE) staining. Aquaporin-3 (AQP3) expression in the colon was analyzed by immunofluorescence, quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blot, respectively.

RESULTS

Compared with the model group, oral administration of LD and HD ESP prevented body weight loss and inhibited diarrhea after 2-week treatment (P<0.05). Kidney deterioration was impeded, and the histology score in LD and HD ESP groups were 8.2 and 10.5, respectively, which were both higher than those in the model group (P<0.05). In addition, ESP treatment alleviated rat colitis, and HD ESP significantly improved the AQP3 positive staining intensity in the colon tissue compared with the model group. The result from Western blot revealed that AQP3 protein synthesis in colon tissue of LD and HD ESP groups increased by 2.1- and 5.9-fold compared with the model group (P<0.05). qRT-PCR result showed that AQP3 gene expression in the HD ESP group was also up-regulated by 2.5-fold normalized to the model group (P<0.05).

CONCLUSION

ESP extract effectively alleviates the symptoms of PSYD and relieves PSYD-induced diarrhea by improving AQP3 synthesis in the colon.

摘要

目的

探讨二至丸(ESP)是否能缓解脾肾阳虚(PSYD)腹泻大鼠模型的症状,并探讨其抗腹泻机制。

方法

75 只 Sprague-Dawley 大鼠采用随机数字表法分为 5 组,包括对照组、阳性组、模型组、低剂量(LD)ESP 组和高剂量(HD)ESP 组,每组 15 只。除对照组外,其余大鼠均根据其病理和病因诱导 PSYD 腹泻。阳性组、LD 和 HD ESP 组大鼠分别给予参苓白术丸()、LD(1.05 g/kg)或 HD(3.50 g/kg)ESP 石油醚提取物,每天 1 次,连续 2 周。测量大鼠体重变化和腹泻指数。通过苏木精-伊红(HE)染色评估肾脏组织学评分。采用免疫荧光、定量逆转录-聚合酶链反应(qRT-PCR)和 Western blot 分别分析结肠中水通道蛋白 3(AQP3)的表达。

结果

与模型组相比,2 周治疗后,口服 LD 和 HD ESP 可防止体重减轻并抑制腹泻(P<0.05)。肾脏恶化受到抑制,LD 和 HD ESP 组的组织学评分分别为 8.2 和 10.5,均高于模型组(P<0.05)。此外,ESP 治疗可缓解大鼠结肠炎,与模型组相比,HD ESP 可显著改善结肠组织中 AQP3 的阳性染色强度。Western blot 结果显示,与模型组相比,LD 和 HD ESP 组结肠组织中 AQP3 蛋白合成增加了 2.1-和 5.9 倍(P<0.05)。qRT-PCR 结果显示,与模型组相比,HD ESP 组 AQP3 基因表达也上调了 2.5 倍(P<0.05)。

结论

ESP 提取物通过提高结肠中 AQP3 的合成,有效缓解 PSYD 症状,缓解 PSYD 引起的腹泻。

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