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. 2022 Mar 29;27(7):2218.
doi: 10.3390/molecules27072218.

Effects of Abelmoschus manihot Flower Extract on Enhancing Sexual Arousal and Reproductive Performance in Zebrafish

Chi-Chang Chang  1   2 , Jer-Yiing Houng  3   4 , Wei-Hao Peng  1 , Tien-Wei Yeh  5 , Yun-Ya Wang  5 , Ya-Ling Chen  2 , Tzu-Hsien Chang  2 , Wei-Chin Hung  6   7 , Teng-Hung Yu  6   7
Affiliations

Effects of Abelmoschus manihot Flower Extract on Enhancing Sexual Arousal and Reproductive Performance in Zebrafish

Chi-Chang Chang et al. Molecules. .

Abstract

The flower of Abelmoschus manihot L. is mainly used for the treatment of chronic kidney diseases, and has been reported to have bioactivities such as antioxidant, anti-inflammatory, antiviral, and antidepressant activities. This study used wild-type adult zebrafish as an animal model to elucidate the potential bioactivity of A. manihot flower ethanol extract (AME) in enhancing their sexual and reproductive functions. Zebrafish were fed AME twice a day at doses of 0.2%, 1%, and 10% for 28 days, and were then given the normal feed for an additional 14 days. The hormone 17-β estradiol was used as the positive control. Sexual behavioral parameters such as the number of times males chased female fish, the production of fertilized eggs, and the hatching rate of the fertilized eggs were recorded at days 0.33, 7, 14, 21, 28, and 42. The expression levels of sex-related genes—including lhcgr, ar, cyp19a1a, and cyp19a1b—were also examined. The results showed that the chasing number, fertilized egg production, and hatching rate were all increased with the increase in the AME treatment dose and treatment time. After feeding with 1% and 10% AME for 28 days, the chasing number in the treated group as compared to the control group increased by 1.52 times and 1.64 times, respectively; the yield of fertilized eggs increased by 1.59 times and 2.31 times, respectively; and the hatching rate increased by 1.26 times and 1.69 times, respectively. All three parameters exhibited strong linear correlations with one another (p < 0.001). The expression of all four genes was also upregulated with increasing AME dose and treatment duration. When feeding with 0.2%, 1%, and 10% AME for 28 days, the four sex-related genes were upregulated at ranges of 1.79−2.08-fold, 2.74−3.73-fold, and 3.30−4.66-fold, respectively. Furthermore, the effect of AME was persistent, as the promotion effect continued after the treatment was stopped for at least two weeks. The present findings suggest that AME can enhance the endocrine system and may improve libido and reproductive performance in zebrafish.

Keywords: Abelmoschus manihot; gene expression; reproductive performance; sexual arousal; zebrafish.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Effects of the supplementation with AME on the chasing number between the male and female zebrafish: Each dataset is represented by five independent repeated experiments. Statistical differences compared to the vehicle group were tested using one-way ANOVA. Levels of significance are expressed as ** p < 0.01 and *** p < 0.001.
Figure 2
Figure 2
Effects of the supplementation of AME on the fertilized egg production. Each dataset is represented by five independent repeated experiments. Statistical differences compared to the vehicle group were tested using one-way ANOVA. Levels of significance are expressed as * p < 0.05, ** p < 0.01, and *** p < 0.001.
Figure 3
Figure 3
Effects of the supplementation with AME on the hatching rate of the fertilized eggs. Each dataset is represented by five independent repeated experiments. Statistical differences compared to the vehicle group were tested using one-way ANOVA. Levels of significance are expressed as ** p < 0.01 and *** p < 0.001.
Figure 4
Figure 4
Effects of the supplementation with AME (A: Female, B: Male) on the body weight of zebrafish during 42-day cultivation. At each sampling time, five male or female adult fish were weighed.
Figure 5
Figure 5
Effects of AME stimulation on the expression of sex-related genes in zebrafish: (A) female, lhcgr in ovaries; (B) female, cyp19a1a in the brain; (C) male, ar in the testes; (D) male, cyp19a1b in the brain. Five adult zebrafish were used for each experimental group at each sampling time. Statistical differences in gene expression between the AME-supplemented-diet group and the vehicle group were tested using one-way ANOVA, and levels of significance are denoted as * p < 0.05, ** p < 0.01, and *** p < 0.001.
Figure 6
Figure 6
HPLC chromatograms of AME: Identified components—rutin, hyperoside, isoquercitrin, myricetin, and quercetin. The peak of the internal standard (naringin) is denoted as “IS”.
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