红景天苷对不育症大鼠睾丸生精细胞增殖凋亡的影响

张常银; 陶众杰; 朱晨杰

doi:10.16766/j.cnki.issn.1674-4152.002448

红景天苷对不育症大鼠睾丸生精细胞增殖凋亡的影响

doi: 10.16766/j.cnki.issn.1674-4152.002448

平湖市中医院泌尿外科, 浙江平湖 314200

基金项目:

浙江省中医药科技计划项目 2019ZB126

详细信息

通讯作者:
张常银，E-mail：xi2356ong@163.com

中图分类号: R256.56 R329.28
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出版历程
- 收稿日期: 2021-05-04
- 网络出版日期: 2022-09-05

Effect of salidroside on the proliferation and apoptosis of testicular spermatogenic cells in infertile rats

Department of Urology, Pinghu Hospital of Traditional Chinese Medicine, Pinghu, Zhejiang 314200, China

摘要

摘要: 目的探讨红景天苷对不育症大鼠睾丸生精细胞增殖凋亡的影响。方法从60只大鼠中按随机数字表法取15只大鼠为对照组(A组)，剩余45只建立不育症模型并根据随机数字表法分为B组、C组、D组，各15只。C组、D组分别以50、100 mg/kg红景天苷灌胃，A组、B组等量生理盐水灌胃。检测血清睾酮、卵泡刺激素、黄体生成素；酶联免疫吸附法(ELISA)检测超氧化物歧化酶(SOD)、丙二醛(MDA)；苏木精-伊红(HE)染色法检测生精细胞形态学；免疫组化法测定增殖细胞核抗原(PCNA)；TUNEL法检测生精细胞凋亡率；Western Blotting法检测脂肪酸合成酶(Fas)、脂肪酸合成酶配体(Fasl)、半胱氨酸蛋白酶-3(caspase-3)蛋白相对表达量。结果与A组比较，B组、C组、D组SOD降低，B组＜C组＜D组(均P＜0.05)；与A组比较，B组、C组、D组MDA及Fas、Fasl、caspase-3蛋白相对表达量升高，B组＞C组＞D组(均P＜0.05)。与B组比较，D组睾丸细胞结构、生精小管萎缩、生精细胞脱落改善更显著。A组睾丸生精细胞PCNA百分率为(41.23±5.46)%，高于B组、C组、D组的(20.13±3.21)%、(27.64±3.25)%、(35.11±4.56)%，B组＜C组＜D组(均P＜0.05)。A组睾丸组织生精细胞凋亡率为(2.23±0.03)%，低于B组、C组、D组的(10.03±1.12)%、(7.94±0.89)%、(4.07±0.52)%，B组＞C组＞D组(均P＜0.05)。结论红景天苷可调节不育症大鼠睾丸生精细胞增殖凋亡，机制可能与下调Fas/Fasl信号通路有关。
- 不育症大鼠 /
- 增殖 /
- 凋亡 /
- 红景天苷
Abstract: Objective To investigate the effect of salidroside on the proliferation and apoptosis of spermatogenic cells in infertile rats. Methods Fifteen rats were Selected from 60 rats as group A, and 45 infertility model rats were randomly divided into groups B, C and D, with 15 rats in each group. Groups C and D were given 50 and 100 mg/kg of salidroside via gavage, and groups A and B were given the same amount of normal saline. Serum testosterone, follicle-stimulating hormone, and luteinising hormone were detected. Superoxide dismutase (SOD) and malondialdehyde (MDA) were detected by enzyme-linked immunosorbent assay. Haematoxylin and eosin (HE) staining was used to detect the morphology of spermatogenic cells. Proliferating cell nuclear antigen (PCNA) was detected by immunohistochemistry, and the apoptosis rate of spermatogenic cells was detected by TUNEL assay. Western blotting was used to detect the relative expression of fatty acid synthase (Fas), fatty acid synthase ligand (Fasl) and caspase-3. Results Compared with that in group A, the SOD levels in groups B, C and D were decreased, and the order was follows: group B < group C < group D (all P < 0.05). Compared with those in group A, the relative expression levels of MDA, Fas, Fasl, and caspase-3 protein in groups B, C and D were increased, and the order was as follows: group B>group C>group D (all P < 0.05). Compared with group B, the structure of testicular cells, atrophy of seminiferous tubules, and exfoliation of spermatogenic cells in group D were significantly improved. The percentage of PCNA in spermatogenic cells of group A was (41.23±5.46)%, which was higher than that in group B (20.13±3.21)%, group C [(27.64±3.25)%] and group D [(35.11±4.56)%], all P < 0.05. The apoptosis rate of spermatogenic cells in group A was (2.23±0.03)%, which was lower than that in group B [(10.03±1.12)%], group C [(7.94±0.89)%] and group D [(4.07±0.52)%], all P < 0.05. Conclusion Salidroside can regulate the proliferation and apoptosis of spermatogenic cells in infertile rats. The mechanism may be related to the downregulation of the Fas/Fasl signalling pathway.
- Infertility rats /
- Proliferation /
- Apoptosis /
- Salidroside

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图 1 各组大鼠睾丸生精细胞形态比较(HE染色，×400)

注：A为对照组(A组)，B为模型组(B组)，C为红景天苷低剂量组(C组)，D为红景天苷高剂量组(D组)。

Figure 1. Morphological comparison of spermatogenic cells in testis of rats in each group (HE staining, ×400)

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图 2 各组大鼠睾丸生精细胞中PCNA百分率(免疫组化，×400)

注：A为对照组(A组)，B为模型组(B组)，C为红景天苷低剂量组(C组)，D为红景天苷高剂量组(D组)。

Figure 2. Percentage of PCNA in testicular spermatogenic cells of rats in each group (Immunohistochemistry, ×400)

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图 3 各组大鼠睾丸生精细胞凋亡率(TUNEL染色，×400)

注：A为对照组(A组)，B为模型组(B组)，C为红景天苷低剂量组(C组)，D为红景天苷高剂量组(D组)。

Figure 3. Apoptosis rate of testicular spermatogenic cells in each group (TUNEL staining, ×400)

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图 4 各组大鼠Fas、Fasl、caspase-3蛋白表达情况

注：A组为对照组，B组为模型组，C组为红景天苷低剂量组，D组为红景天苷高剂量组。

Figure 4. The protein expression of Fas, Fasl and caspase-3 in each group

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表 1 各组大鼠血清睾酮、卵泡刺激素、黄体生成素水平比较(x±s)

Table 1. Comparison of serum testosterone, follicle stimulating hormone and luteinizing hormone levels in each group(x±s)

组别	只数	睾酮(nmol/L)	卵泡刺激素(IU/L)	黄体生成素(IU/L)
A组	15	189.54±32.52	2.02±0.05	7.35±0.26
B组	15	60.25±10.25^a	2.28±0.06^a	6.02±0.35^a
C组	15	118.25±11.28^ab	2.17±0.08^ab	6.85±0.36^ab
D组	15	127.05±12.75^abc	2.10±0.07^abc	7.11±0.31^abc
F值		115.693	41.925	48.354
P值		＜0.001	＜0.001	＜0.001
注：与A组比较，^aP＜0.05；与B组比较，^bP＜0.05；与C组比较，^cP＜0.05。

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表 2 各组大鼠血清SOD活性、MDA含量比较(x±s)

Table 2. Comparison of SOD activity and MDA content in serum of rats in each group(x±s)

组别	只数	SOD(U/mL)	MDA(nmol/mg)
A组	15	156.23±19.21	0.51±0.06
B组	15	90.35±10.11^a	1.56±0.21^a
C组	15	115.31±12.32^ab	1.16±0.18^ab
D组	15	136.59±13.98^abc	0.81±0.11^abc
F值		58.734	133.406
P值		＜0.001	＜0.001
注：与A组比较，^aP＜0.05；与B组比较，^bP＜0.05；与C组比较，^cP＜0.05。

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表 3 各组大鼠Fas、Fasl、caspase-3蛋白相对表达量比较(x±s)

Table 3. The relative expressions of Fas, Fasl and caspase-3 in each group were compared(x±s)

组别	只数	Fas	Fasl	caspase-3
A组	8	0.25±0.04	0.22±0.04	0.14±0.03
B组	8	0.87±0.10^a	0.90±0.14^a	0.75±0.11^a
C组	8	0.59±0.09^ab	0.61±0.08^ab	0.48±0.08^ab
D组	8	0.40±0.07^abc	0.41±0.05^abc	0.34±0.07^abc
F值		92.997	89.905	86.508
P值		＜0.001	＜0.001	＜0.001
注：与A组比较，^aP＜0.05；与B组比较，^bP＜0.05；与C组比较，^cP＜0.05。

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