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目的:比较二神丸方剂中的两味中药炮制前、后对脾肾阳虚泄泻模型大鼠全血T淋巴细胞亚群和环磷酸腺苷(cAMP)与环磷酸鸟苷(cGMP)比值(cAMP/cGMP)失衡的调节作用,为阐明二神丸中药物炮制增效机制提供实验依据。方法:将50只SD雄性大鼠随机分为正常、模型、阳性对照参芩白术丸2 g·kg-1组,二神丸生品(生补骨脂+生肉豆蔻)3.5 g·kg-1组、二神丸炮制品(盐补骨脂+煨肉豆蔻)3.5 g·kg-1组共5组,通过复合因素造脾肾阳虚泄泻模型4周,并按组给药2周,分别从股动脉和眼眶后静脉丛取全血和血清,用流式细胞仪检测全血各组样本T淋巴细胞亚群CD3+,CD3+CD4+,CD3+CD8+的百分比;用酶联免疫吸附法(ELISA)测定血清样本中环磷酸腺苷(cAMP)和环磷酸鸟苷(cGMP)的水平。结果:3个给药组均能显著提高模型动物血清中cAMP/cGMP和全血CD3+,CD3+CD4+百分比,而CD3+CD8+百分比则显著降低,且炮制品二神丸组对CD3+,CD3+CD4+,CD3+CD8+百分比和cAMP/cGMP的调节作用明显强于生品二神丸组。结论:复方二神丸中药物经过炮制后能显著恢复脾肾阳虚泄泻模型大鼠全血CD3+CD4+,CD3+CD8+(P<0.01)和血清cAMP/cGMP(P<0.01)的水平,提示二神丸中的两味中药炮制后增强疗效可能与其增强脾肾阳虚泄泻模型大鼠对T细胞网络平衡(CD3+,CD3+CD4+,CD3+CD8+)和cAMP/cGMP平衡的调节作用有关。
OBJECTIVE: To compare the T lymphocyte subsets, cAMP and cGMP ratio in whole blood of spleen and kidney yang deficiency diarrhea model before and after the preparation of Liangwei traditional Chinese medicine (cAMP / cGMP) imbalance in regulating the role of Ershen pill in order to clarify the mechanism of synergy to provide experimental evidence. Methods: Fifty male Sprague-Dawley rats were randomly divided into two groups: normal, model and positive control group. The rats in 2 g · kg-1 group were treated with Shengkai Baizhu pill (3.5 g · kg -1) Group, group of two tablets of Ershen pill (salt psoralen + simmered nutmeg) 3.5 g · kg-1 group of 5 groups, made by the combined factors of spleen and kidney yang deficiency diarrhea model for 4 weeks, and the group administered for 2 weeks, Whole blood and serum were collected from the femoral artery and retro-orbital vein respectively. The percentages of CD3 +, CD3 +, CD4 + and CD3 + CD8 + of T lymphocyte subsets were detected by flow cytometry. The percentage of CD3 + ELISA) was used to determine levels of cAMP and cGMP in serum samples. Results: The percentages of cAMP / cGMP and whole blood CD3 +, CD3 + and CD4 + in serum of model animals were significantly increased and the percentage of CD3 + CD8 + , The regulation of CD3 + CD8 + percentage and cAMP / cGMP were significantly stronger than that of Erzheng pill group. Conclusion: After the preparation of Fufang Er Shen Piao, the levels of CD3 + CD4 +, CD3 + CD8 + (P <0.01) and cAMP / cGMP (P <0.01) It is suggested that the enhancement effect of the two herbs in Er Shen Wan after processing could be related to the regulation of balance of T cell network (CD3 +, CD3 + CD4 +, CD3 + CD8 +) and cAMP / cGMP balance in the spleen-kidney yang diarrhea model rats .