肾宁颗粒制备工艺探讨及药效学研究
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摘要
目的：优选肾宁颗粒制备工艺中黄芪、益母草与川芎的提取工艺；观察肾宁颗粒急性毒性；考察肾宁颗粒对大鼠阳离子化牛血清白蛋白(C-BSA)肾炎模型的治疗作用。
方法：采用正交试验法，以干膏收率、黄芪甲苷和黄芪多糖含量为指标，对黄芪的水煎提取工艺进行了优选；以干膏收率、盐酸水苏碱含量为指标考察益母草与川芎水煎提取工艺；以80％乙醇浸出物得率、盐酸水苏碱含量为指标考察益母草与川芎水煎提取液的醇沉条件。
雄性Wistar大鼠56只，随机分为正常组、模型组、肾宁颗粒高、中、低剂量组、肾复康组、泼尼松组，每组8只。以阳离子化牛血清白蛋白(C-BSA)造成大鼠膜性肾炎模型，第二周末起每周测定一次24h尿蛋白，实验结束时测定血浆白蛋白、肾功能、前列腺素等，并做病理检查，观察各组治疗效果。
结果：黄芪的最佳提取工艺为水煎3次，每次1h，每次加8倍量水；益母草与川芎的最佳提取工艺为水煎3次，每次2h，每次加12倍量水，将所得的水煎液浓缩至1：1.5后，加乙醇醇沉，使醇沉液中乙醇浓度为50％，醇沉时间24h。肾宁颗粒小鼠的最大耐受量为240g·Kg~(-1)。与模型组比较，肾宁颗粒可降低C-BSA肾炎模型大鼠尿蛋白含量，以高剂量组起效最早，效果最显著。肾宁颗粒剂可升高模型大鼠血浆白蛋白含量，降低血浆肌酐和尿素氮含量，能降低C-BSA肾炎大鼠血浆血栓素B_2(TXB_2)／6-酮基-前列腺素F_(1α)(6-Keto-PGF_(1α))值，对肾功能有保护作用。
结论：所建立的黄芪、益母草与川芎提取工艺对主要有效成分提取率高，实用性好。肾宁颗粒口服的安全性很大。肾宁颗粒高、中剂量组可明显降低膜性肾炎大鼠尿蛋白含量，高剂量组可升高大鼠血浆白蛋白水平，在改善肾功能方面作用同肾复康作用相似，优于泼尼松组。
Objective To optimize the conditions for the extractions of Radix Astragali, Herba leonuri and Rhizoma Chuanxiong in the preparation of Shenning Granues.To investigate effects of Shenning Granues on nephrotic rats.
Methord By orthogonal experimental design, the water decocting techniques were optimized with the dried water decocting rates and contents of starchy trine hydrochloride etc .And the condition of alcohol sedimentation were optimized with the extraction rate of 80% alcohol and contents of starchy trine hydrochloride .
Adult male wistar rats were randomly devided into normal control group, pathological pattern group, high-dosage shenning group, middle-dosage group, low-dosage group, Shenfukang group and Prednisone group. Rats with membranous glomerulonephritis were induced by injection of cation bovine serum albumin into tail vein. The rats were treated respectively for 32 days by gavage. Urinary protein was observed once a week after 2 weeks. After 7weeks, plasma albumin, renal histology, parameters of renal function and prostaglandins were examined. In addition TXB2/6-Keto-PGF1. in serum were also examined.
Result The optimized decocting condition for Radix Astragalan were to boil the Radix Astragalan for 3 times, each time for 1 hour and add water as much as 8 times. The optimized decocting condition for Herba leonuri and Rhizoma Chuanxiong were to boil Herba leonuri and Rhizoma Chuanxiong for 3 times, each time for 2 hours and add water as much as 12 times, the best condition in the technology of alcohol sedimentation were to condensed the decocting liquid to l:1.5(g:ml) and sediment for 24 hours in 50% alcohol.
The urinary protein, serum cretinine level, BUN and TXB2/6-Keto-PGF1. can be reduced in Shenning groups compared with pathological pattern group. Plasma albumin was significantly increased in high-dosage Shenning group.
Conclusion The processes of extraction of Radix Astragalan , Herba leonuri and Rhizoma Chuanxiong are feasible .
High-dosage and middle-dosage Shenning can significantly decrease urinary protein in rats with membranous glomerulonephritis. High-dosage Shenning can increase the plasma albumin level in rats with membranous glomerulonephritis.
方法：采用正交试验法，以干膏收率、黄芪甲苷和黄芪多糖含量为指标，对黄芪的水煎提取工艺进行了优选；以干膏收率、盐酸水苏碱含量为指标考察益母草与川芎水煎提取工艺；以80％乙醇浸出物得率、盐酸水苏碱含量为指标考察益母草与川芎水煎提取液的醇沉条件。
雄性Wistar大鼠56只，随机分为正常组、模型组、肾宁颗粒高、中、低剂量组、肾复康组、泼尼松组，每组8只。以阳离子化牛血清白蛋白(C-BSA)造成大鼠膜性肾炎模型，第二周末起每周测定一次24h尿蛋白，实验结束时测定血浆白蛋白、肾功能、前列腺素等，并做病理检查，观察各组治疗效果。
结果：黄芪的最佳提取工艺为水煎3次，每次1h，每次加8倍量水；益母草与川芎的最佳提取工艺为水煎3次，每次2h，每次加12倍量水，将所得的水煎液浓缩至1：1.5后，加乙醇醇沉，使醇沉液中乙醇浓度为50％，醇沉时间24h。肾宁颗粒小鼠的最大耐受量为240g·Kg~(-1)。与模型组比较，肾宁颗粒可降低C-BSA肾炎模型大鼠尿蛋白含量，以高剂量组起效最早，效果最显著。肾宁颗粒剂可升高模型大鼠血浆白蛋白含量，降低血浆肌酐和尿素氮含量，能降低C-BSA肾炎大鼠血浆血栓素B_2(TXB_2)／6-酮基-前列腺素F_(1α)(6-Keto-PGF_(1α))值，对肾功能有保护作用。
结论：所建立的黄芪、益母草与川芎提取工艺对主要有效成分提取率高，实用性好。肾宁颗粒口服的安全性很大。肾宁颗粒高、中剂量组可明显降低膜性肾炎大鼠尿蛋白含量，高剂量组可升高大鼠血浆白蛋白水平，在改善肾功能方面作用同肾复康作用相似，优于泼尼松组。
Objective To optimize the conditions for the extractions of Radix Astragali, Herba leonuri and Rhizoma Chuanxiong in the preparation of Shenning Granues.To investigate effects of Shenning Granues on nephrotic rats.
Methord By orthogonal experimental design, the water decocting techniques were optimized with the dried water decocting rates and contents of starchy trine hydrochloride etc .And the condition of alcohol sedimentation were optimized with the extraction rate of 80% alcohol and contents of starchy trine hydrochloride .
Adult male wistar rats were randomly devided into normal control group, pathological pattern group, high-dosage shenning group, middle-dosage group, low-dosage group, Shenfukang group and Prednisone group. Rats with membranous glomerulonephritis were induced by injection of cation bovine serum albumin into tail vein. The rats were treated respectively for 32 days by gavage. Urinary protein was observed once a week after 2 weeks. After 7weeks, plasma albumin, renal histology, parameters of renal function and prostaglandins were examined. In addition TXB2/6-Keto-PGF1. in serum were also examined.
Result The optimized decocting condition for Radix Astragalan were to boil the Radix Astragalan for 3 times, each time for 1 hour and add water as much as 8 times. The optimized decocting condition for Herba leonuri and Rhizoma Chuanxiong were to boil Herba leonuri and Rhizoma Chuanxiong for 3 times, each time for 2 hours and add water as much as 12 times, the best condition in the technology of alcohol sedimentation were to condensed the decocting liquid to l:1.5(g:ml) and sediment for 24 hours in 50% alcohol.
The urinary protein, serum cretinine level, BUN and TXB2/6-Keto-PGF1. can be reduced in Shenning groups compared with pathological pattern group. Plasma albumin was significantly increased in high-dosage Shenning group.
Conclusion The processes of extraction of Radix Astragalan , Herba leonuri and Rhizoma Chuanxiong are feasible .
High-dosage and middle-dosage Shenning can significantly decrease urinary protein in rats with membranous glomerulonephritis. High-dosage Shenning can increase the plasma albumin level in rats with membranous glomerulonephritis.
引文
1．张鸣和，柴锡庆．内科学．北京：北京医科大学出版社，1997，279～288
2．王海燕．肾脏病学．北京：人民卫生出版社，1996，697～702
3．王自敏．中西医临床肾病学．北京：中国中医药出版社 1997，156～161
4．叶任高．中医对肾小球肾炎的治疗现状[J]．中华肾脏病杂志，1990，6(4)：236～238
5．黎磊石，关天俊，刘志红，等．4298例成年人肾小球疾病病理类型及流行病学特点[J]．肾脏病与透析肾移植杂志，1997，6(2)：103～105．
6．钱漪，许树梧．实用药学计算．湖南：湖南科学技术出版社，1983，347～350
7．谢秀琼．中药新制剂开发与应用．北京：人民卫生出版社，2000，779
8．中国药典(一部)，2000，249．
9．张贤华，郝季涛．TLC测定肾宁合剂中黄芪甲苷的含量[J]．中国现代应用药学杂志，1999，16(1)：65
10．中国药典(一部)，2000，64．
11．张玲，于宗渊，李国宝，等．七种含益母草中成药中水苏碱的含量测定[J]．药物分析杂志，1996，16(3)：181～183
12．张玲，时延增，于宗渊，等．双波长薄层扫描法测定益母草口服液中水苏碱的含量[J]．中国药科大学学报，1996，27(1)：16～18
13．李伯延．植物药有效成分的提取分离．太原：山西高校联合出版社，1993，172．
14．中国药典(一部)，2000，附录63．
15．中药新药申报指南．药理、毒理学
16. Border WA, Ward H, Bachelor RW et al. Induction of membranous nephropathy in rabbits by administration of an exogenous cationic antigen. Demonstration of pathogenic role for electrical charge[J]. J Clin invest, 1982, 69: 451.
17．施新猷．现代医学实验动物学．北京：人民军医出版社，2000，334．
18．李锐，曹建宏，周玖瑶．肾复康对大鼠实验性慢性肾炎的作用[J]．广州中医药大学学报，1999，16(3)：205～208
19．阎汝南，王静竹，刘舒平，等．HPLC法测定黄芪中黄芪甲苷的含量[J]．中国中药杂
志，1998，23(7)：398．
20．孟景春，等．黄芪为治慢性肾炎蛋白尿的良药[J] 江苏中医杂志，1995，16(5)：316．
21. Lidenos E A, Andres G A, Dum M J. Glomerular prostaglandin and thromboxane synthesis in rat nephritic serum nephritis[J]. J Clin invest, 1983, 172: 1439.
2．王海燕．肾脏病学．北京：人民卫生出版社，1996，697～702
3．王自敏．中西医临床肾病学．北京：中国中医药出版社 1997，156～161
4．叶任高．中医对肾小球肾炎的治疗现状[J]．中华肾脏病杂志，1990，6(4)：236～238
5．黎磊石，关天俊，刘志红，等．4298例成年人肾小球疾病病理类型及流行病学特点[J]．肾脏病与透析肾移植杂志，1997，6(2)：103～105．
6．钱漪，许树梧．实用药学计算．湖南：湖南科学技术出版社，1983，347～350
7．谢秀琼．中药新制剂开发与应用．北京：人民卫生出版社，2000，779
8．中国药典(一部)，2000，249．
9．张贤华，郝季涛．TLC测定肾宁合剂中黄芪甲苷的含量[J]．中国现代应用药学杂志，1999，16(1)：65
10．中国药典(一部)，2000，64．
11．张玲，于宗渊，李国宝，等．七种含益母草中成药中水苏碱的含量测定[J]．药物分析杂志，1996，16(3)：181～183
12．张玲，时延增，于宗渊，等．双波长薄层扫描法测定益母草口服液中水苏碱的含量[J]．中国药科大学学报，1996，27(1)：16～18
13．李伯延．植物药有效成分的提取分离．太原：山西高校联合出版社，1993，172．
14．中国药典(一部)，2000，附录63．
15．中药新药申报指南．药理、毒理学
16. Border WA, Ward H, Bachelor RW et al. Induction of membranous nephropathy in rabbits by administration of an exogenous cationic antigen. Demonstration of pathogenic role for electrical charge[J]. J Clin invest, 1982, 69: 451.
17．施新猷．现代医学实验动物学．北京：人民军医出版社，2000，334．
18．李锐，曹建宏，周玖瑶．肾复康对大鼠实验性慢性肾炎的作用[J]．广州中医药大学学报，1999，16(3)：205～208
19．阎汝南，王静竹，刘舒平，等．HPLC法测定黄芪中黄芪甲苷的含量[J]．中国中药杂
志，1998，23(7)：398．
20．孟景春，等．黄芪为治慢性肾炎蛋白尿的良药[J] 江苏中医杂志，1995，16(5)：316．
21. Lidenos E A, Andres G A, Dum M J. Glomerular prostaglandin and thromboxane synthesis in rat nephritic serum nephritis[J]. J Clin invest, 1983, 172: 1439.