不同中药药对调节氢化可的松药源性证候小鼠肾上腺皮质功能的研究
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摘要
目的研究不同治法中药配伍调节肾上腺皮质功能的作用。方法小鼠ig给予1.65 mg/(kg·d)氢化可的松14 d诱发药源性证候,同步给予助阳散寒(附子-肉桂)、滋阴泻火(知母-黄柏)、补气生津(人参-黄芪)、补血益精(熟地-山茱萸)4种中药药对干预,检测小鼠体质量、抓力、腋温、旷场活动度、躯体不同部位红外温度等表征信息;取脾脏、胸腺称定质量并计算脏器指数;运用ELISA法检测血清皮质酮、促肾上腺皮质激素(ACTH)、肾上腺素和去甲肾上腺素含量;采用实时荧光定量PCR技术检测肾上腺类固醇合成急性调节蛋白(StAR)、胆固醇侧链裂解酶(Cyp11a1)、21-羟化酶(Cyp21a1)、11β-羟化酶1(Cyp11b1)和11β-羟化酶2(Cyp11b2)、酪氨酸羟化酶(Th)、多巴胺脱羧酶(Ddc)、多巴胺β-羟化酶(Dbh)、苯乙胺-N-甲基转移酶(Pnmt)基因表达;运用Western blotting方法检测肾上腺低密度脂蛋白受体(LDLR)、B族I型清道夫受体(SR-BI)、StAR蛋白表达。结果 4种治法对氢化可的松造成的小鼠体质量下降和体温下降效果不显著,附子-肉桂组小鼠抓力显著高于模型组(P<0.05)。与模型组比较,知母-黄柏组小鼠血清皮质酮含量显著升高(P<0.01),StAR、Cyp11b1、Cyp11b2基因表达水平显著升高(P<0.05、0.01),SR-BI、StAR蛋白表达水平均显著升高(P<0.01)。结论滋阴泻火治法寒性药对(知母-黄柏)对小剂量氢化可的松诱发的药源性证候小鼠肾上腺皮质功能的纠正作用最佳。
Objective To study the effects of different Chinese materia medica combinations on the regulation of adrenal function.Methods Mice with 1.65 mg/(kg·d) hydrocortisone for 14 d to induce drug-induced syndrome was treated synchronously with assissting yang and dissipating cold(Aconm Lateralis Radix Praeparaia-Cinnamomi Cortex), nourishing yin and reducing fire(Anemarrhenae Rhizoma-Phellodendri Chinensis Cortex), tonifying qi and promoting fluid production(Ginseng Radix et Rhizoma-Astragali Radix), and tonifying blood and benefiting spirit(Rehmanniae Radix Praeparata-Corni Fructus). The body mass of mice was dynamically observed every day, and the characteristic information such as body mass, holding power, axillary temperature,activity degrees of open field, and infrared temperature were detected by the experimental methodology of mice syndrome differentiation; The mice were sacrificed whose spleen, thymus was weighed and the organ index was calculated. Serum corticosterone,ACTH, adrenaline, and noradrenaline were measured by ELISA. The gene expressions of Star, Cyp11 a1, Cyp21 a1, Cyp11 b1,Cyp11 b2, Dbh, Ddc, Pnmt, and Th in the adrenal were detected by real-time fluorescent quantitative PCR. The protein's expressions of LDLR, SRB1, and StAR were detected by Western blotting. Results The effects of four different treatments on the reduction of body weight and body temperature caused by hydrocortisone were not significant. The holding power of mice in Aconm Lateralis Radix Praeparaia-Cinnamomi Cortex group was significantly higher than that of hydrocortisone group(P < 0.05). Compared with the hydrocortisone group, the serum corticosterone content of mice in Anemarrhenae Rhizoma-Phellodendri Chinensis Cortex group increased significantly(P < 0.01); The genes expression of Star, Cyp11 b1, and Cyp11 b2 was significantly up-regulated(P < 0.05, 0.01);The proteins expression of SRBI and StAR were significantly up-regulated. Conclusion The nourishing yin and purging fire treatment group(Anemarrhenae Rhizoma-Phellodendri Chinensis Cortex) is the best to correct and protect the adrenal cortex function in mice with hydrocortisone induced syndrome at low dose.
Objective To study the effects of different Chinese materia medica combinations on the regulation of adrenal function.Methods Mice with 1.65 mg/(kg·d) hydrocortisone for 14 d to induce drug-induced syndrome was treated synchronously with assissting yang and dissipating cold(Aconm Lateralis Radix Praeparaia-Cinnamomi Cortex), nourishing yin and reducing fire(Anemarrhenae Rhizoma-Phellodendri Chinensis Cortex), tonifying qi and promoting fluid production(Ginseng Radix et Rhizoma-Astragali Radix), and tonifying blood and benefiting spirit(Rehmanniae Radix Praeparata-Corni Fructus). The body mass of mice was dynamically observed every day, and the characteristic information such as body mass, holding power, axillary temperature,activity degrees of open field, and infrared temperature were detected by the experimental methodology of mice syndrome differentiation; The mice were sacrificed whose spleen, thymus was weighed and the organ index was calculated. Serum corticosterone,ACTH, adrenaline, and noradrenaline were measured by ELISA. The gene expressions of Star, Cyp11 a1, Cyp21 a1, Cyp11 b1,Cyp11 b2, Dbh, Ddc, Pnmt, and Th in the adrenal were detected by real-time fluorescent quantitative PCR. The protein's expressions of LDLR, SRB1, and StAR were detected by Western blotting. Results The effects of four different treatments on the reduction of body weight and body temperature caused by hydrocortisone were not significant. The holding power of mice in Aconm Lateralis Radix Praeparaia-Cinnamomi Cortex group was significantly higher than that of hydrocortisone group(P < 0.05). Compared with the hydrocortisone group, the serum corticosterone content of mice in Anemarrhenae Rhizoma-Phellodendri Chinensis Cortex group increased significantly(P < 0.01); The genes expression of Star, Cyp11 b1, and Cyp11 b2 was significantly up-regulated(P < 0.05, 0.01);The proteins expression of SRBI and StAR were significantly up-regulated. Conclusion The nourishing yin and purging fire treatment group(Anemarrhenae Rhizoma-Phellodendri Chinensis Cortex) is the best to correct and protect the adrenal cortex function in mice with hydrocortisone induced syndrome at low dose.
引文
[1]潘志强.“药源性证候”的新生学术问题与思考[J].上海中医药杂志, 2018, 52(6):5-8.
[2]潘志强,梁龙龙,王晓敏,等.氢化可的松诱发虚证模型小鼠的证候特征再评价及肾上腺皮质功能变化研究[J].上海中医药杂志, 2017, 51(8):13-20.
[3]万姜维,刘锋.中医药治疗糖皮质激素副作用研究进展[J].河南中医, 2016, 36(5):914-917.
[4]钟天飞,徐莉,李海昌.中医角度浅析糖皮质激素与上火的关系[J].山西中医学院学报, 2016, 17(2):12-14.
[5]田丽,张伟.从少火壮火论浅析中药对糖皮质激素的增效减毒作用[J].长春中医药大学学报,2019,35(1):13-15.
[6]吴斌.糖皮质激素副作用的中医药研究进展[J].时珍国医国药, 2010, 21(3):719-721.
[7]章伟.糖皮质激素治疗中的中医思维与对策[J].医学理论与实践, 2017, 30(18):2683-2684.
[8]范永升.对中医药减轻激素副作用的思考与实践[A]//全国第十二届中西医结合风湿病学术会议论文汇编[C].天津:中国中西医结合学会, 2014.
[9]方肇勤,潘志强,汤伟昌,等.小鼠“四诊工作站”构建与操作标准探讨[J].上海中医药大学学报,2006,20(1):42-46.
[10]方肇勤,潘志强,付晓伶,等.小鼠四诊采集项目标准的建议[J].中国中医基础医学杂志,2005,11(9):692-694.
[11]方肇勤,潘志强,卢文丽,等.大鼠、小鼠常见证候计量化辨证方法的建立及其评价[J].中国中医基础医学杂志, 2007, 13(7):502-505.
[12]杨洪雁,苏双良,付晶,等.低密度脂蛋白受体基因研究进展[J].东北农业大学学报, 2013, 44(3):141-146.
[13]常冠楠,徐新,张社兵.B类I型清道夫受体的研究进展[J].中西医结合心脑血管病杂志,2010,8(9):1126-1129.
[14]李娟,栾新红.类固醇合成急性调节蛋白的生物学特性及作用研究概况[J].动物医学进展,2016,37(1):110-113.
[2]潘志强,梁龙龙,王晓敏,等.氢化可的松诱发虚证模型小鼠的证候特征再评价及肾上腺皮质功能变化研究[J].上海中医药杂志, 2017, 51(8):13-20.
[3]万姜维,刘锋.中医药治疗糖皮质激素副作用研究进展[J].河南中医, 2016, 36(5):914-917.
[4]钟天飞,徐莉,李海昌.中医角度浅析糖皮质激素与上火的关系[J].山西中医学院学报, 2016, 17(2):12-14.
[5]田丽,张伟.从少火壮火论浅析中药对糖皮质激素的增效减毒作用[J].长春中医药大学学报,2019,35(1):13-15.
[6]吴斌.糖皮质激素副作用的中医药研究进展[J].时珍国医国药, 2010, 21(3):719-721.
[7]章伟.糖皮质激素治疗中的中医思维与对策[J].医学理论与实践, 2017, 30(18):2683-2684.
[8]范永升.对中医药减轻激素副作用的思考与实践[A]//全国第十二届中西医结合风湿病学术会议论文汇编[C].天津:中国中西医结合学会, 2014.
[9]方肇勤,潘志强,汤伟昌,等.小鼠“四诊工作站”构建与操作标准探讨[J].上海中医药大学学报,2006,20(1):42-46.
[10]方肇勤,潘志强,付晓伶,等.小鼠四诊采集项目标准的建议[J].中国中医基础医学杂志,2005,11(9):692-694.
[11]方肇勤,潘志强,卢文丽,等.大鼠、小鼠常见证候计量化辨证方法的建立及其评价[J].中国中医基础医学杂志, 2007, 13(7):502-505.
[12]杨洪雁,苏双良,付晶,等.低密度脂蛋白受体基因研究进展[J].东北农业大学学报, 2013, 44(3):141-146.
[13]常冠楠,徐新,张社兵.B类I型清道夫受体的研究进展[J].中西医结合心脑血管病杂志,2010,8(9):1126-1129.
[14]李娟,栾新红.类固醇合成急性调节蛋白的生物学特性及作用研究概况[J].动物医学进展,2016,37(1):110-113.