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纸质出版日期:2018
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张晨曦, 王永辉, 许凯霞, 等. 不同补益方剂对老龄小鼠生精细胞凋亡的影响[J]. 中国实验方剂学杂志, 2018,24(8):148-153.
ZHANG Chen-xi, WANG Yong-hui, XU Kai-xia, et al. Effect of Different Nourishing Formulas on Spermatogenic Cell Apoptosis of Aging Mice[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(8): 148-153.
张晨曦, 王永辉, 许凯霞, 等. 不同补益方剂对老龄小鼠生精细胞凋亡的影响[J]. 中国实验方剂学杂志, 2018,24(8):148-153. DOI: 10.13422/j.cnki.syfjx.20180826.
ZHANG Chen-xi, WANG Yong-hui, XU Kai-xia, et al. Effect of Different Nourishing Formulas on Spermatogenic Cell Apoptosis of Aging Mice[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(8): 148-153. DOI: 10.13422/j.cnki.syfjx.20180826.
目的:探讨不同补益方剂左归丸、右归丸、六味地黄丸、金匮肾气丸、五子衍宗丸、八珍汤对老龄小鼠生精细胞凋亡的影响。方法:将80只15月龄BALB/c小鼠随机分为老年组、左归丸组、右归丸组、六味地黄丸组、金匮肾气丸组、五子衍宗丸组、八珍汤组以及阳性药物(左卡尼汀)组,每组10只,另设10只2月龄BALB/c小鼠为青年组,各给药组按中药含生药2.5 g·kg-1分别灌胃给予相应药物,左卡尼汀组肌肉注射给药45 mg·kg-1,每天1次,连续4周。于末次给药24 h后将小鼠摘眼球取血,分离血浆、血清,利用酶联免疫吸附法(ELISA)测定小鼠血中丙二醛(MDA),一氧化氮合酶(NOS),促红细胞生成素(EPO),超氧化物歧化酶(SOD)以及外周血促卵泡生成激素(FSH),促黄体生成激素(LH),睾酮(T)的含量;采用免疫组化法检测各组小鼠睾丸组织Ki-67的表达;采用TUNEL法检测各组小鼠生精细胞凋亡情况。结果:与青年组比较,老年组小鼠MDA,NOS,EPO水平明显上升(P<0.05);与老年组比较,各治疗组小鼠血清MDA,NOS,EPO水平明显下降(P<0.05),除五子衍宗丸组和六味地黄丸组外,各治疗组小鼠血清SOD水平上升(P<0.05)。与青年组比较,老年组小鼠血清FSH明显升高及LH,T含量明显下降(P<0.05);与老年组比较,五子衍宗丸组小鼠血清FSH明显下降及LH,T含量明显升高(P<0.05)。免疫组化法检测结果显示,与青年组比较,老年组小鼠睾丸组织内Ki-67表达水平明显下降(P<0.05);与老年组比较,左卡尼汀组、六味地黄丸组、八珍汤组以及五子衍宗丸组小鼠睾丸组织Ki-67表达水平升高(P<0.05)。TUNEL法检测结果显示,与青年组比较,老年组小鼠生精细胞凋亡率明显上升(P<0.05);与老年组比较,各治疗组小鼠生精细胞凋亡率均明显下降,以五子衍宗丸下降率最显著(P<0.05)。结论:6个不同的补益方剂对老龄小鼠生精细胞凋亡都有一定的抑制作用,其中以五子衍宗丸效果最佳。
Objective: To the effect of different nourishing formulas Zuoguiwan
Youguiwan
Liuwei Dihuangwan
Jinkui Shenqiwan
Wuzi Yanzong Wan and Bazhentang on spermatogenic cell apoptosis of aging mice. Method: A total of 80 15-month-old aging mice were divided into 8 groups
aging group
Zuoguiwan group
Youguiwan group
Liuwei Dihuangwan group
Jinkui Shenqiwan group
Wuzi Yanzong Wan group
Bazhentang group and levocarnitine group
with 10 in each group. Another 10 2-month-old young mice were selected as control group. All of the drug groups were given the corresponding drugs by gavage at the dosage of 2.5 g · kg-1
and levocarnitine group was intramuscularly injected with the drug at the dosage of 45 mg · kg-1. All mice were administered once a day
for 4 weeks in a row. Twenty-four hours after the last administration
blood was collected from the mice by removing their eyes
and serum was separate to detect the contents of malondialdehyde(MDA)
nitric oxide synthase(NOS)
erythropoietin(EPO)
superoxide dismutase(SOD)
follicle stimulating hormone(FSH)
leuteinizing hormone(LH) and testosterone(T). Then
the apoptosis of spermatogenic cells and the expression of Ki-67 in testis were detected in each group. Result: Compared with control group
the contents of MDA
NOS and EPO in aging group increased
with no significant difference in SOD(P<0.05). Compared with aging group
the contents of MDA
NOS and EPO in treatment groups decreased
while their SOD increased (except for Wuzi Yanzong Wan group and Liuwei Dihuangwan)(P<0.05). Compared with control group
aging group's FSN increased
while LH and T significantly decreased(P<0.05). Compared with aging group
the content of FSN significantly decreased
whereas LH and T increased(P<0.05). Based on the immunohistochemical method
compared with control group
aging group's Ki-67 expression in testis decreased
Compared with aging group
Liuwei Dihuangwan group
Wuzi Yanzong Wan group
Bazhentang group and levocarnitine group showed an increase in the expression of Ki-67 in testis(P<0.05). On the basis of Tunel method
compared with control group
the apoptosis rate of spermatogenic cells significantly increased in aging group
while that of all of the treatment groups decreased
especially in the Wuzi Yanzong Wan group(P<0.05). Conclusion: All of the six different nourishing formulas have an inhibitory effect on the apoptosis of spermatogenic cells of aging mice
with the best efficacy in Wuzi Yanzong Wan.
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