基于阴虚大鼠模型的玉竹有效部位筛选及其理化性质分析
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摘要
目的探讨玉竹干预阴虚大鼠模型的有效部位,分离纯化玉竹多糖(Polygonatumodoratum Polysaccharides, POP),并对其基本理化性质进行分析。方法制备玉竹水总提取物(Ⅰ)、醇沉上清液(Ⅱ)和醇沉淀(Ⅲ)3个部位,大鼠腹腔注射三碘甲状腺原氨酸(T3),每日1次,连续1周,建立阴虚大鼠模型。检测各部位对阴虚大鼠体质量及体温变化的影响,采用Elisa法检测大鼠血清游离三碘甲状腺原氨酸(fT3)、游离四碘甲状腺原氨酸(f T4)、血浆环磷酸腺苷(cAMP)、环磷酸鸟苷(cGMP)水平。采用DEAE-52纤维素柱层析对POP进行分离纯化,并对各均一多糖进行理化性质分析。结果玉竹各部位均能显著增加阴虚大鼠的体质量,降低阴虚大鼠体温,对阴虚大鼠血清fT3、f T4、血浆cAMP、cGMP水平紊乱调节作用显著(P<0.01),且所含多糖的醇沉淀(Ⅲ)部位效果最佳。POP中分离得到1种中性多糖(POP1)和2种酸性多糖(POP2、POP3),其中POP1含量最高。结论以中性多糖为主的玉竹多糖具有最佳滋阴药效,该研究为玉竹多糖的进一步开发提供了基础。
Objective To investigate the active fractions of Polygonatum odoratum in a rat model of Yin deficiency, to extract and purify Polygonatum odoratum polysaccharides(POPs), and to analyze their physicochemical properties. Methods Three fractions were prepared from Polygonatum odoratum, including the whole-plant aqueous extract(POP1), and supernatant(POP2) and precipitate(POP3) from ethanol precipitation. The rat model of Yin deficiency was established by intraperitoneal injection of triiodothyronine(T3) once a day for 1 week. The effect of the three fractions on the changes in body weight and temperature of the rats with Yin deficiency was evaluated. The levels of free triiodothyronine(FT3) and free tetraiodothyronine(FT4) in serum, and cyclic adenosine monophosphate(cAMP) and cyclic guanosine monophosphate(cGMP) in plasma were measured by ELISA. POPs were extracted and purified by DEAE-cellulose-52 chromatography, and the physicochemical properties of each homogeneous polysaccharide were analyzed. Results All the fractions showed a significant effect in increasing the body weight, reducing the temperature, and regulating the serum levels of FT3 and FT4 and plasma levels of cAMP and cGMP in the rats with Yin deficiency(P<0.01), with the best effect observed in POP 3. One type of neutral polysaccharide(POP1) and two types of acidic polysaccharides(POP2 and POP3) were extracted from POPs, among which the content of POP1 was highest. Conclusion The bestYin-nourishment effect is observed in the neutral polysaccharide-dominated Polygonatum odoratum polysaccharide. This study provides a basis for the further development of Polygonatum odoratum polysaccharides.
Objective To investigate the active fractions of Polygonatum odoratum in a rat model of Yin deficiency, to extract and purify Polygonatum odoratum polysaccharides(POPs), and to analyze their physicochemical properties. Methods Three fractions were prepared from Polygonatum odoratum, including the whole-plant aqueous extract(POP1), and supernatant(POP2) and precipitate(POP3) from ethanol precipitation. The rat model of Yin deficiency was established by intraperitoneal injection of triiodothyronine(T3) once a day for 1 week. The effect of the three fractions on the changes in body weight and temperature of the rats with Yin deficiency was evaluated. The levels of free triiodothyronine(FT3) and free tetraiodothyronine(FT4) in serum, and cyclic adenosine monophosphate(cAMP) and cyclic guanosine monophosphate(cGMP) in plasma were measured by ELISA. POPs were extracted and purified by DEAE-cellulose-52 chromatography, and the physicochemical properties of each homogeneous polysaccharide were analyzed. Results All the fractions showed a significant effect in increasing the body weight, reducing the temperature, and regulating the serum levels of FT3 and FT4 and plasma levels of cAMP and cGMP in the rats with Yin deficiency(P<0.01), with the best effect observed in POP 3. One type of neutral polysaccharide(POP1) and two types of acidic polysaccharides(POP2 and POP3) were extracted from POPs, among which the content of POP1 was highest. Conclusion The bestYin-nourishment effect is observed in the neutral polysaccharide-dominated Polygonatum odoratum polysaccharide. This study provides a basis for the further development of Polygonatum odoratum polysaccharides.
引文
[1]杨慧洁,杨世海,张海弢,等.玉竹化学成分、药理作用研究进展及开发利用现状[J].人参研究,2012,24(3):40-45.
[2]丁登峰,向大雄,刘韶,等.玉竹多糖的提取及其对链脉佐菌素诱导糖尿病大鼠血糖的影响[J].中南药学,2005,3(4):222-224.
[3]张轩铭,王冬梅,王瑾,等.不同产地玉竹黄酮提取物体外抗氧化活性研究[J].西北植物学报,2011,31(3):628-631.
[4]吴国学.玉竹对小鼠免疫抑制调节作用的研究[J].中国医学创新,2013,10(9):13-14.
[5]胡方林,刘仙菊,易法银,等.益气滋阴方调节甲亢模型大鼠甲状腺激素的实验研究[J].现在中西医结合杂志,2008,17(25):3902-3903.
[6]黄江荣,李祥华,张家均,等.六味地黄丸对甲状腺功能亢进肾阴虚型小鼠cAMP、cGMP含量和Na+, K+-ATP酶活性的影响[J].中药药理与临床,2011,27(6):1-3.
[7]傅万山,丁伯平,杨解.六味地黄丸对甲亢型肾阴虚大鼠滋阴作用的研究[J].中国实验方剂学杂志,2001,7(5):16-18.
[8]毛水龙,周卫民,郑怡健,等.六味地黄丸对甲状腺素所致甲亢大鼠脂肪组织UCP2、骨骼肌UCP3mRNA表达的影响[J].浙江中西医结合杂志,2012,22(11):849-852.
[9]张智敏,胡玉珍,熊苏慧,等.玉竹水提醇沉上清液对甲亢阴虚型大鼠干预的GC-MS代谢组学研究[J].中国药学杂志,2017,52(23):2092-2097.
[10]王强,李盛钰,杨帆,等.玉竹中性多糖的分离纯化及单糖组成分析[J].食品科学,2010,31(15):100-102.
[11]屈凯,曲锋,樊雪红,等.手心温度与阴虚证、五心烦热症的相关性临床研究[J].山东中医药大学学报,2010,34(2):134-135.
[12] STRICH D, KARAVANI G, EDRI S, et al. TSH enhancement of FT4 to FT3 conversion is age dependent[J]. Eur J Endocrinol,2016, 175(1):49-54.
[13] WANG Q, REN X J, YAO S L, et al. Clinical observation on the endocrinal and immune functions in subjects with yin-deficiency constitution[J]. Chin J Integr Med, 2010, 16(1):28-32.
[14]占阮娟,陈素红,吕圭源,等.三味甘凉归肺经中药对肺阴虚模型大鼠cAMP、cGMP的影响[J].中药药理与临床,2012,28(6):85-87.
[15]王楠楠,戴明珠,徐俞悦,等.铁皮石斛花对肾上腺皮质激素致肾阴虚模型小鼠的影响[J].中药药理与临床,2017,33(1):116-119.
[16]吴柳花,吕圭源,李波,等.黄精对长期超负荷游泳致阴虚内热模型大鼠的作用研究[J].中国中药杂志,2014,39(10):1886-1891.
[17]林昌松,李宁,徐强,等.知柏养阴汤对阴虚模型大鼠糖脂代谢的影响[J].时珍国医国药,2011,22(10):2404-2405.
[18]张凡,徐珊,刘蓬蓬,等.黄柏不同炮制品对甲亢型肾阴虚模型大鼠甲状腺和肾上腺皮质功能的影响[J].中国药房,2017,28(1):27-30.
[19]刘婷,候连兵,候毅成.甲状腺片诱导的甲亢阴虚证大鼠模型的制备[J].中药材,2008,31(9):1402-1404.
[2]丁登峰,向大雄,刘韶,等.玉竹多糖的提取及其对链脉佐菌素诱导糖尿病大鼠血糖的影响[J].中南药学,2005,3(4):222-224.
[3]张轩铭,王冬梅,王瑾,等.不同产地玉竹黄酮提取物体外抗氧化活性研究[J].西北植物学报,2011,31(3):628-631.
[4]吴国学.玉竹对小鼠免疫抑制调节作用的研究[J].中国医学创新,2013,10(9):13-14.
[5]胡方林,刘仙菊,易法银,等.益气滋阴方调节甲亢模型大鼠甲状腺激素的实验研究[J].现在中西医结合杂志,2008,17(25):3902-3903.
[6]黄江荣,李祥华,张家均,等.六味地黄丸对甲状腺功能亢进肾阴虚型小鼠cAMP、cGMP含量和Na+, K+-ATP酶活性的影响[J].中药药理与临床,2011,27(6):1-3.
[7]傅万山,丁伯平,杨解.六味地黄丸对甲亢型肾阴虚大鼠滋阴作用的研究[J].中国实验方剂学杂志,2001,7(5):16-18.
[8]毛水龙,周卫民,郑怡健,等.六味地黄丸对甲状腺素所致甲亢大鼠脂肪组织UCP2、骨骼肌UCP3mRNA表达的影响[J].浙江中西医结合杂志,2012,22(11):849-852.
[9]张智敏,胡玉珍,熊苏慧,等.玉竹水提醇沉上清液对甲亢阴虚型大鼠干预的GC-MS代谢组学研究[J].中国药学杂志,2017,52(23):2092-2097.
[10]王强,李盛钰,杨帆,等.玉竹中性多糖的分离纯化及单糖组成分析[J].食品科学,2010,31(15):100-102.
[11]屈凯,曲锋,樊雪红,等.手心温度与阴虚证、五心烦热症的相关性临床研究[J].山东中医药大学学报,2010,34(2):134-135.
[12] STRICH D, KARAVANI G, EDRI S, et al. TSH enhancement of FT4 to FT3 conversion is age dependent[J]. Eur J Endocrinol,2016, 175(1):49-54.
[13] WANG Q, REN X J, YAO S L, et al. Clinical observation on the endocrinal and immune functions in subjects with yin-deficiency constitution[J]. Chin J Integr Med, 2010, 16(1):28-32.
[14]占阮娟,陈素红,吕圭源,等.三味甘凉归肺经中药对肺阴虚模型大鼠cAMP、cGMP的影响[J].中药药理与临床,2012,28(6):85-87.
[15]王楠楠,戴明珠,徐俞悦,等.铁皮石斛花对肾上腺皮质激素致肾阴虚模型小鼠的影响[J].中药药理与临床,2017,33(1):116-119.
[16]吴柳花,吕圭源,李波,等.黄精对长期超负荷游泳致阴虚内热模型大鼠的作用研究[J].中国中药杂志,2014,39(10):1886-1891.
[17]林昌松,李宁,徐强,等.知柏养阴汤对阴虚模型大鼠糖脂代谢的影响[J].时珍国医国药,2011,22(10):2404-2405.
[18]张凡,徐珊,刘蓬蓬,等.黄柏不同炮制品对甲亢型肾阴虚模型大鼠甲状腺和肾上腺皮质功能的影响[J].中国药房,2017,28(1):27-30.
[19]刘婷,候连兵,候毅成.甲状腺片诱导的甲亢阴虚证大鼠模型的制备[J].中药材,2008,31(9):1402-1404.