中药复方对衰老及肾阳虚小鼠激素水平及肾功能的影响
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摘要
为研究中药复方对D-半乳糖联合氢化可的松注射液致小鼠衰老及肾阳虚证的影响,选取60只5周龄昆明雄性小鼠随机分为5组,除空白组外,其余4组小鼠1 d～40 d每日颈背部皮下注射D-半乳糖(200 mg/kg)溶液,第41天～第50天,持续后肢注射氢化可的松注射液为小鼠衰老及肾阳虚,在此基础上给予低(0.80 g/kg)、中(1.60 g/kg)、高(3.20 g/kg)剂量的中药复方,用药周期为68 d,同时空白组、模型组灌胃等剂量的生理盐水。试验期间观察小鼠精神状态,记录小鼠体重和肛温。用药结束后,处死小鼠,分离血清,检测小鼠血清中T3、T4、TSH、ACTH、COR含量,摘取肾脏,进行切片HE染色,肾组织匀浆中检测NO含量,血清中检测CRE、BUN水平。结果显示,与空白组相比,模型组小鼠体重增长缓慢,活动减少,精神沉郁,肛温降低,血清中T3、T4、ACTH含量显著降低(P<0.01),TSH显著升高(P<0.01),COR无显著性变化;肾组织匀浆中NO升高(P<0.01),血清中CRE、BUN含量显著升高(P<0.01),HE染色显示肾小球萎缩,肾小管上皮细胞变性、坏死,肾小囊上皮细胞肿胀。中药复方可提高小鼠体重、肛温、血清中T3、T4、ACTH含量、降低肾组织匀浆中NO含量及血清中CRE、BUN水平,减轻肾脏组织病理损害。结果表明,中药复方对D-半乳糖及氢化可的松致衰老及肾阳虚小鼠内分泌调节紊乱、肾脏受损具有一定保护作用。
To investigate the protective effect of herbal formula on geriatric and Kidney-Yang deficiency mice induced by D-galactose combining with hydrocortisone injection,sixty 5-week-old KM male mice were randomly divided into 5 groups.Except for the control group,the other 4 groups were injected subcutaneously with D-galactose(200 mg/kg d) solution daily for 1-40 d,and then injected with hydrocortisone during 41-50 d at hind limb to set up geriatric and Kidney-Yang deficiency mice model.Three groups of the mice model were given with low(0.80 g/kg d),medium(1.60 g/kg),and high(3.20 g/kg) dose of herbal formula respectively for successive 68 days.The control group and model group were given with equal doses of normal saline.The mental activities of all the mice were observed along the experiment,the body weight,and rectal temperature of the mice were collected.At the end of experiment,the mice were sacrificed and the serum was separated.The levels of T3,T4,TSH,ACTH,and COR in the serum were measured.The kidney organs were removed and HE staining was performed.The content of NO was detected in the renal homogenate, and CRE and BUN levels were detected in the serum.The results showed that compared with the control group, the mice in the model group had slower growth rate,reduced activities, decreased rectal temperature.The T3,T4,ACTH levels in the serum were significantly decreased(P<0.01),and TSH content was increased significantly(P<0.01),while there is no significant changes for COR content. The level of NO in renal homogenates was increased(P<0.01),and the levels of CRE and BUN in serum were significantly increased(P<0.01).The histologic study showed glomerular atrophy,degeneration and necrosis of renal tubular epithelial cells and swelling of renal cyst epithelial cells.After adding the herbal formula,the body weight, rectal temperature,serum T3,T4 and ACTH levels were increased significantly,while the content of NO in renal homogenate and levels of CRE and BUN in serum were decreased,the renal morphology significantly improved.These results suggested that Herbal formula has protective effects on endocrine disorder and kidney damage in geriatric and Kidney-Yang deficiency mice induced by D-galactose and hydrocortisone injection.
To investigate the protective effect of herbal formula on geriatric and Kidney-Yang deficiency mice induced by D-galactose combining with hydrocortisone injection,sixty 5-week-old KM male mice were randomly divided into 5 groups.Except for the control group,the other 4 groups were injected subcutaneously with D-galactose(200 mg/kg d) solution daily for 1-40 d,and then injected with hydrocortisone during 41-50 d at hind limb to set up geriatric and Kidney-Yang deficiency mice model.Three groups of the mice model were given with low(0.80 g/kg d),medium(1.60 g/kg),and high(3.20 g/kg) dose of herbal formula respectively for successive 68 days.The control group and model group were given with equal doses of normal saline.The mental activities of all the mice were observed along the experiment,the body weight,and rectal temperature of the mice were collected.At the end of experiment,the mice were sacrificed and the serum was separated.The levels of T3,T4,TSH,ACTH,and COR in the serum were measured.The kidney organs were removed and HE staining was performed.The content of NO was detected in the renal homogenate, and CRE and BUN levels were detected in the serum.The results showed that compared with the control group, the mice in the model group had slower growth rate,reduced activities, decreased rectal temperature.The T3,T4,ACTH levels in the serum were significantly decreased(P<0.01),and TSH content was increased significantly(P<0.01),while there is no significant changes for COR content. The level of NO in renal homogenates was increased(P<0.01),and the levels of CRE and BUN in serum were significantly increased(P<0.01).The histologic study showed glomerular atrophy,degeneration and necrosis of renal tubular epithelial cells and swelling of renal cyst epithelial cells.After adding the herbal formula,the body weight, rectal temperature,serum T3,T4 and ACTH levels were increased significantly,while the content of NO in renal homogenate and levels of CRE and BUN in serum were decreased,the renal morphology significantly improved.These results suggested that Herbal formula has protective effects on endocrine disorder and kidney damage in geriatric and Kidney-Yang deficiency mice induced by D-galactose and hydrocortisone injection.
引文
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[16] 崔松焕,张宇,金春爱,等.鹿茸保健口服液对氢化可的松所致小鼠肾阳虚症的影响[C]//首届中国鹿业发展大会暨中国畜牧业协会鹿业分会,内蒙古包头:2010.
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[20] 徐翠姣,陈国祥,余亮,等.附子黄芪胶囊治疗慢性肾小球肾炎脾肾阳虚型临床研究[J].内蒙古中医药,2014,33(22):1-2.
[21] 朱大年.生理学[M].北京:人民卫生出版社,2013:388-396.
[22] 戴冰,杨梦琳,张嘉妮,等.从HPT轴角度探讨桂枝-附子在金匮肾气汤中温补肾阳的作用机理[J].中医药信息,2018(2):43-46.
[23] 金蓉家,杨元宵,邢桂英,等.肾气丸对肾阳虚大鼠下丘脑-垂体-甲状腺轴的调节作用初探[J].浙江中医杂志,2013,48(5):370-371.
[24] 岳仁宋,吴施国,谭毅明,等.参附注射液对甲状腺机能减退症肾阳虚型小鼠血清甲状腺激素的影响[J].中国中医急症,2005(11):1094-1095.
[25] 张磊,廖成彬,黄榕波,等.右归丸调节肾阳虚垂体-靶腺轴功能的动态变化[J].广东药学院学报,2016,32(1):102-106.
[26] 闵友江,姚海华,程立红.艾条悬灸肾俞、关元对肾阳虚大鼠垂体-肾上腺轴和垂体-甲状腺轴的影响[J].上海针灸杂志,2016,35(12):1469-1472.
[27] 汪玉梅,高洁,李政木,等.基于响应面分析法的肾气丸组方优化研究[J].新中医,2016,48(8):292-296.
[28] Li X,Wang J,Zhang H,et al.Renoprotective effect of low-molecular-weight sulfated polysaccharide from the seaweed Laminaria japonica on glycerol-induced acute kidney injury in rats[J].Int J Biol Macromol,2017,95:132-137.
[29] 施明珠,李有贵,钟石,等.北冬虫夏草对氢化可的松诱导的老龄小鼠肾损伤的保护作用[J].现代药物与临床,2009,24(3):160-163.
[30] 王朝霞.公驼宝克对肾阳虚大鼠的影响[D].内蒙古呼和浩特:内蒙古农业大学,2011.
[31] 胡胜林.艾灸对肾阳虚模型大鼠组织形态学及血清ACTH、CORT影响的研究[D].湖南长沙:湖南中医药大学,2015.
[2] 安方玉,颜春鲁,刘永琦,等.黄芪多糖对镉染毒大鼠肾损伤的保护作用[J].中国临床药理学杂志,2018,34(1):49-52.
[3] 吴秋玉,郑远鹏,林志谦,等.黄芪多糖作为饲料添加剂及免疫增强剂在畜牧生产中的应用[J].中国饲料,2018(2):30-33.
[4] 吕德包,覃艳艳.在日粮中添加“黄芪多糖”预防仔猪早期断奶应激综合征的试验报告[J].广西畜牧兽医,2018,34(1):21-23.
[5] 敬舒,刘健,李宁,等.北五味子木脂素对D-半乳糖致衰小鼠学习记忆能力的影响[J].北华大学学报:自然科学版,2016,17(6):745-749.
[6] 王馨雅,高慧,王晓婷,等.五味子不同炮制品对大鼠肾阳虚的改善作用研究[J].现代药物与临床,2018,33(1):10-14.
[7] 刘志勇,徐铁山,顾丽红,等.日粮添加五味子对文昌鸡生长性能和肉质性状的影响[J].中国家禽,2017,39(22):24-27.
[8] 陈辉,陈洁,孙锋,等.金匮肾气丸对大鼠肾脏纤维化的影响[J].中医学报,2015,30(7):996-999.
[9] 宋卫中,刘蔚,张保国.金匮肾气丸(汤)在老年疾病中的应用[J].时珍国医国药,2016,27(2):439-441.
[10] 张玲红.金匮肾气丸加减治疗脾肾阳虚型糖尿病肾病的疗效及对肾功能的影响研究[J].中西医结合心血管病电子杂志,2018,6(2):169-170.
[11] 石娜,苏洁,杨正标,等.白术多糖对D-半乳糖致衰老模型小鼠的抗氧化作用[J].中国新药杂志,2014,23(5):577-581.
[12] 邱洪,刘景会,骆鹏飞,等.金匮肾气丸水提液对肾阳虚雄鼠生长性能及其睾丸形态结构的影响[J].黑龙江畜牧兽医,2014(11):4-7.
[13] 郝青,周焕娇,刘建民.D-半乳糖和氢化可的松诱导老年阳虚证大鼠模型的建立[J].湖北中医药大学学报,2014,16(5):9-12.
[14] 武莹莹,李梦云,宋格格,等.激动素对衰老模型小鼠免疫力的影响研究[J].畜牧与饲料科学,2016,37(3):18-21.
[15] 戴冰,张嘉妮,杨梦琳,等.氢化可的松致肾虚证小鼠模型的建立及相关指标的评价[J].中国实验动物学报,2017,25(1):70-73.
[16] 崔松焕,张宇,金春爱,等.鹿茸保健口服液对氢化可的松所致小鼠肾阳虚症的影响[C]//首届中国鹿业发展大会暨中国畜牧业协会鹿业分会,内蒙古包头:2010.
[17] 刘梦云,韩旭.肾虚与衰老机理中西医研究概况[J].辽宁中医药大学学报,2011,13(11):108-111.
[18] 孙筱婷,刘宣,李琦.肾虚证动物模型研究概况[J].中华中医药杂志,2017,32(2):659-662.
[19] 吴骁,王光,李彦,等.黄芪注射液对D-半乳糖所致衰老小鼠的免疫、衰老和血脂指标的影响[J].实验动物科学,2011,28(6):7-9.
[20] 徐翠姣,陈国祥,余亮,等.附子黄芪胶囊治疗慢性肾小球肾炎脾肾阳虚型临床研究[J].内蒙古中医药,2014,33(22):1-2.
[21] 朱大年.生理学[M].北京:人民卫生出版社,2013:388-396.
[22] 戴冰,杨梦琳,张嘉妮,等.从HPT轴角度探讨桂枝-附子在金匮肾气汤中温补肾阳的作用机理[J].中医药信息,2018(2):43-46.
[23] 金蓉家,杨元宵,邢桂英,等.肾气丸对肾阳虚大鼠下丘脑-垂体-甲状腺轴的调节作用初探[J].浙江中医杂志,2013,48(5):370-371.
[24] 岳仁宋,吴施国,谭毅明,等.参附注射液对甲状腺机能减退症肾阳虚型小鼠血清甲状腺激素的影响[J].中国中医急症,2005(11):1094-1095.
[25] 张磊,廖成彬,黄榕波,等.右归丸调节肾阳虚垂体-靶腺轴功能的动态变化[J].广东药学院学报,2016,32(1):102-106.
[26] 闵友江,姚海华,程立红.艾条悬灸肾俞、关元对肾阳虚大鼠垂体-肾上腺轴和垂体-甲状腺轴的影响[J].上海针灸杂志,2016,35(12):1469-1472.
[27] 汪玉梅,高洁,李政木,等.基于响应面分析法的肾气丸组方优化研究[J].新中医,2016,48(8):292-296.
[28] Li X,Wang J,Zhang H,et al.Renoprotective effect of low-molecular-weight sulfated polysaccharide from the seaweed Laminaria japonica on glycerol-induced acute kidney injury in rats[J].Int J Biol Macromol,2017,95:132-137.
[29] 施明珠,李有贵,钟石,等.北冬虫夏草对氢化可的松诱导的老龄小鼠肾损伤的保护作用[J].现代药物与临床,2009,24(3):160-163.
[30] 王朝霞.公驼宝克对肾阳虚大鼠的影响[D].内蒙古呼和浩特:内蒙古农业大学,2011.
[31] 胡胜林.艾灸对肾阳虚模型大鼠组织形态学及血清ACTH、CORT影响的研究[D].湖南长沙:湖南中医药大学,2015.