Human Amnion Mesenchymal Stem Cells Improve Testicular Steroidogenesis in Aging Mice

doi:10.12280/gjszjk.20210138

Abstract

Abstract:

Objective: To explore whether human amnion mesenchymal stem cells(hAMSC) can improve testicular steroidogenesis in the aging mice. Methods: 31 male mice aged 36-weeks as the model of aging males were randomly divided into 5 groups, with 3 mice in the normal saline control group and 7 mice in the other four groups. Different doses of hAMSC (3.4×10⁶ cells/kg, 6.7×10⁶ cells/kg, 1.4×10⁷ cells/kg) were injected through the tail vein, and the 1% human serum albumin group was used as the treatment control. And the normal saline was used as the blank control. Seven mice aged 10 weeks were used as the young control. The mice were injected four times and once a week. After five weeks of recovery, both testes were taken. The histopathological change was observed by HE staining. The expressions of StAR and 17β-HSD in testicular tissues were detected by immunoblotting. The expressions of STEM121 and CD105 were also detected by immunofluorescence. The epididymis was taken to evaluate sperm count, sperm motility rate and sperm morphology. The level of testosterone was tested by chemiluminescence immune assay.Results: Compared with the young control, the aged mice in the blank control and the treatment control had the increased body weight, the decreased testicular mass-to-weight ratio, the increased fat around the testes (all P<0.01), and the decreased sperm motility rate and normal morphological sperm rate (bothP<0.05). There were no significant differences in sperm density and the expressions of StAR and 17β-HSD protein in testicular tissues between the aging control and the young control (allP>0.05). After five weeks of the hAMSC treatment, the immunofluorescence images showed that there were many cells with the positive expressions of STEM121 and CD105 in the interstitial area of testis. There were no significant differences in the expression of StAR in testis among the treatment groups. The expression of 17β-HSD in the high-dose group was increased than that in the treatment-control group, and the level of testosterone was also increased (bothP<0.05). The histopathology showed that the cell layers of seminiferous tubules were increased in the medium dose group and the high dose group than those in the treatment control group (P<0.05).Conclusions: The preliminary results indicate that hAMSC has a protective effect on testicular aging and can improve testicular steroidogenesis in the aging mice.

Key words: Mesenchymal stromal cells, Amnion, Aging, Fertility, Mesenchymal stem cell transplantation, Human amnion mesenchymal stem cells, Testicular aging, Spermatogenic function, Steroidogenesis

ZHANG Yue-xin, LIU Han-wen, SHI Chen-nan, NING Song, ZHOU Jing, LI Chu-yu, YANG Xiao-yu, QIN Lian-ju, LIU Jia-yin, HU Yan-qiu, CUI Yu-gui. Human Amnion Mesenchymal Stem Cells Improve Testicular Steroidogenesis in Aging Mice[J]. Journal of International Reproductive Health/Family Planning, 2021, 40(4): 265-271.

Figures/Tables 11

References 21

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组别	n	体质量（g）	睾丸体质比	睾周脂肪质量（g）	血清睾酮（nmol/L）	StAR/ β-actin	17β-HSD/ GAPDH	精子计数（个/mL,×10⁶）	精子活动率（%）	正常形态精子率（%）	生精小管直径（μm）	生精小管细胞层数
年轻对照组	7	24.33±0.33	0.74±0.02	0.33±0.00	46.44±9.05	0.61±0.24	0.49±0.08	5.80±0.59	72.93±8.07	95.00±1.73	146.90±55.48	8.20±1.40
治疗前高龄组	7	37.67±0.88	0.57±0.02	2.42±0.27	12.91±2.25	0.65±0.31	0.44±0.18	5.88±1.58	52.57±9.25	85.43±2.76	190.90±27.07	5.80±1.22
t		14.14	4.86	7.61	3.60	0.23	0.51	0.08	2.87	5.09	3.92	7.09
P		0.00	0.01	0.00	0.01	0.82	0.63	0.94	0.04	0.01	0.00	0.00

组别	n	体质量（g）	睾丸体质比	睾周脂肪质量（g）	血清睾酮（nmol/L）	StAR/ β-actin	17β-HSD/ GAPDH	精子计数（个/mL,×10⁶）	精子活动率（%）	正常形态精子率（%）	生精小管直径（μm）	生精小管细胞层数
年轻对照组	7	24.33±0.33	0.74±0.02	0.33±0.00	46.44±9.05	0.61±0.24	0.49±0.08	5.80±0.59	72.93±8.07	95.00±1.73	146.90±55.48	8.20±1.40
治疗前高龄组	7	37.67±0.88	0.57±0.02	2.42±0.27	12.91±2.25	0.65±0.31	0.44±0.18	5.88±1.58	52.57±9.25	85.43±2.76	190.90±27.07	5.80±1.22
t		14.14	4.86	7.61	3.60	0.23	0.51	0.08	2.87	5.09	3.92	7.09
P		0.00	0.01	0.00	0.01	0.82	0.63	0.94	0.04	0.01	0.00	0.00

组别	n	血清睾酮	StAR/β-actin	17β-HSD/GAPDH
年轻对照组	7	46.44±9.05	0.61±0.10	0.49±0.04
治疗前高龄组	7	12.91±2.25	0.65±0.13	0.44±0.09
生理盐水对照组	3	16.87±3.31*	0.70±0.21	0.51±0.11
F		9.79	0.13	0.27
P		0.01	0.88	0.77

组别	n	血清睾酮	StAR/β-actin	17β-HSD/GAPDH
年轻对照组	7	46.44±9.05	0.61±0.10	0.49±0.04
治疗前高龄组	7	12.91±2.25	0.65±0.13	0.44±0.09
生理盐水对照组	3	16.87±3.31*	0.70±0.21	0.51±0.11
F		9.79	0.13	0.27
P		0.01	0.88	0.77

组别	n	血清睾酮	StAR/β-actin	17β-HSD/GAPDH
生理盐水对照组	3	16.87±3.31	0.70±0.21	0.51±0.11
人血清白蛋白对照组	7	14.85±2.72	0.54±0.18	0.49±0.11
低剂量组	7	18.36±3.41	0.49±0.19	0.48±0.19
中剂量组	7	15.24±1.92	0.55±0.17	0.71±0.10
高剂量组	7	28.51±9.40**	0.84±0.33	0.82±0.09*
F		5.56	1.63	4.53
P		0.01	0.40	0.02