Protective effect of ammonium trichloride tellurate (AS101) on ovarian injury induced by chemotherapy drug doxorubicin in rats

Abstract

Objective: To investigate the protective effect of ammonium trichloride tellurate (AS101) on doxorubicin induced ovarian function damage in rats.

Methods: Eighteen female SD rats were randomly divided into three groups with 6 rats in each group: group A blank control group (0.9% normal saline 0.1 ml intraperitoneally for 21 consecutive days), group B (0.9% normal saline 0.1 ml intraperitoneally for 5 days, doxorubicin 10 mg / kg intraperitoneally on day 6, and continued to be injected with normal saline for 15 days), group C (0.9% normal saline 0.1 ml intraperitoneally for 5 days, doxorubicin 10 mg / kg intraperitoneally on day 6, and continued to be injected with AS101 twice every other day). The contents to be recorded by the operator team include: (1) before operation: ① observe the general state of rats in each group and record the weight of rats. ② Vaginal smears of rats in each group were observed at the same time every day to observe the estrous cycle of rats. (2) After operation: ① observe the general state of rats in each group, read the weight of rats, and record the data Vaginal smears of rats in each group were observed at the same time every day after operation to observe the estrous cycle of rats. ③ At the end of the postoperative observation period, blood samples were taken from the tail of rats to measure the serum levels of estrogen 2 (E2), follicle stimulating hormone (FSH), and anti-M ü llerian hormone (AMH). ④ At the end of the observation period, the animals were sacrificed to observe the development of bilateral ovarian follicles. ⑤ At the end of the observation period, blood was taken from the abdominal aorta of rats after anesthesia, and the upper serum was separated after centrifugation at 3000 rpm. The levels of malondialdehyde (MDA), superoxide dismutase (SOD) and reactive oxygen species (ROS) were measured according to the operation methods of the kit instructions.

Result: The body weight of rats in group B was significantly lower than that in group A, while that in group C was significantly higher than that in group B (P < 0.05). The estrous cycle of rats in group B was disordered, and the estrous cycle of rats in group C was restored regularly (P < 0.05). The serum estradiol(E2) and AMH levels in group C were significantly higher than those in group B, and the FSH level was significantly lower than that in group B. Compared with group B, the number of primary follicles increased and the number of atretic follicles decreased in group C (P < 0.05). The SOD level in group B was lower than that in group A, and that in group C was higher than that in group B (P < 0.05). The levels of MDA and ROS in group B were higher than those in group A (P < 0.05), while those in group C were lower than those in group B (P < 0.05).

Conclusion: AS101 can protect the damage of doxorubicin on ovarian function in rats, and its mechanism is related to its antioxidant effect.

Clinical trial number: Not applicable.

Keywords: AS101; Chemotherapy; Ovarian; Oxidative stress; Reproductive endocrine.

Fig. 1

General state of rats in each groups. (A) Group A (saline)(n = 6); (B) Group B (saline + doxorubicin) (n = 6); (C) Group C (saline + doxorubicin AS101) (n = 6). After collecting blood samples from each group of rats for further analysis, the 6 samples were uniformly measured using the ELISA in a single test. The experiment was repeated 3 times, and since the conclusions were consistent, the dataset with the best results was selected for statistical analysis. Moreover, the data presented in this study (tables and bar charts) represent the mean value of 6 samples per group of rats

Fig. 2

Trends in body weight of rats in each group

Fig. 3

HE staining of vaginal smear cells from normal rat estrous cycles. (A) Proestrus: The duration typically ranges from 12 to 14 h, with nucleated epithelial cells being the predominant cell type observed in rat vaginal smears, along with occasional keratinized cells and leukocytes; (B) Estrus: The duration typically ranges from 25 to 27 h, with rat vaginal smears consisting entirely of anucleate keratinized epithelial cells, occasionally accompanied by a few oval-shaped epithelial cells; (C) Interestrus: The duration typically ranges from 55 to 57 h, with the most notable characteristic of the vaginal smear being the presence of numerous leukocytes and a small number of nucleated epithelial cells; (D) Late estrus: The duration typically ranges from 6 to 8 h, with the vaginal smear displaying a mixture of cell types, including leukocytes, keratinized cells, and nucleated epithelial cells. Blue arrow: unnucleated keratinocyte; Orange arrows: nucleated epithelial cells; Green arrows: white blood cells

Fig. 4

Estrous cycle of rats in each group before intervention

Fig. 5

Estrous cycle of rats in each group after intervention

Fig. 6

Comparison of serum E2 levels of rats in each group after intervention

Fig. 7

Comparison of serum FSH levels of rats in each group after intervention

Fig. 8

Comparison of serum AMH levels of rats in each group after intervention

Fig. 9

Comparison of MDA levels in ovarian tissue of rats after intervention

Fig. 10

Comparison of SOD levels in ovarian tissue of rats in each group after intervention

Fig. 11

Comparison of ROS levels in ovarian tissue of rats in each group after intervention

Fig. 12

HE staining of ovarian tissue in each groups. (A) HE staining of ovarian tissue in Ggroup A; (B) HE staining of ovarian tissue in Group B; (C) HE staining of ovarian tissue in Group C. Black arrow: Primordial follicle. Green arrow: Atretic follicle

Fig. 13

Comparison of the number of primordial follicles in each group after intervention

Fig. 14

Comparison of the number of atretic follicles in each group after intervention