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Journal of Chinese Integrative Medicine ›› 2009, Vol. 7 ›› Issue (6): 532-540.doi: 10.3736/jcim20090608

• Original Experimental Research • Previous Articles     Next Articles

Effects of Bushen Kangshuai Tang in retrieving oxidative stress-induced reproductive defects in Caenorhabditis elegans

Xin-guo Cao, Li-li Hou, Jun-xia Chen, Qin Lu   

  1. Department of Traditional Chinese Medicine, Nanjing Maternity and Child Health Care Hospital, Nanjing Medical University, Nanjing 210004, Jiangsu Province, China
  • Received:2008-09-22 Accepted:2009-04-03 Online:2009-06-20 Published:2009-06-15

Objective

To explore the function of Bushen Kangshuai Tang (BKT), a compound traditional Chinese herbal medicine, in alleviating oxidative stress-induced reproductive defects in organism nematode Caenorhabditis elegans.
Methods

The L4-larvae were cultured with 25%, 50%, 75%, and 100% of BKT with the final concentration of 0.33 g/mL. Ultraviolet irradiation [20 J/(m 2·min)], heat-shock (36 ℃, 2 h), and paraquat treatment (2 mmol/L, 2 h) were exerted as the stresses. Reproduction was assayed by the egg number in uterus, brood size, and generation time.
Results

Ultraviolet irradiation, heat-shock, and paraquat treatment could significantly reduce egg number in uterus and brood size, increase generation time, and suppress activities of catalase and superoxide dismutase of the treated wild-type N2 nematodes. Administration of BKT did not cause toxic or altered effects on reproduction under wild-type background in C. elegans. However, BKT administration at higher concentrations could not only effectively alleviate the reproductive defects induced by ultraviolet irradiation, heat-shock, and paraquat treatment, but also increase the catalase and superoxide dismutase activities suppressed by ultraviolet irradiation, heat-shock, and paraquat treatment as compared with the control. Moreover, administration of BKT at higher concentrations could largely ecover the reproductive defects formed in mev-1 mutant nematodes with elevated oxidative stress.
Conclusion

Oxidative stress can negatively regulate the reproductive process, and the administration of high concentrations of BKT will largely retrieve the oxidative stress-induced reproductive defects in exposed nematodes.

Key words: Bushen Kangshuai Tang, Oxidative stress, Reproduction, Caenorhabditis elegans

Figure 1

Effects of BKT administration on egg number in uterus, brood size and generation time in wild-type N2 nematodes Exposures were performed for 2 h on L4-larva stage nematodes with 25%, 50%, 75%, and 100% of examined BKT diluted with M9 buffer. At least 20 replicates were performed for assay of egg number in uterus, brood size and generation time for statistical purposes. One-way ANOVA followed by a Dunnett's t test was used to determine the significance of the differences between the groups. Data are expressed as x±s, n=30."

Figure 2

Retrieval effects of BKT administration on egg number in uterus, brood size and generation time in UV-irradiated wild-type N2 nematodes Approximately 30 L2-stage larvae were irradiated on NGM plates without food at 20 J/(m2·min). All UV irradiation assays were performed at 20 ℃, and afterwards further maintained at 20 ℃. Exposures were performed for 2 h on L4-larva stage nematodes with 25%, 50%, 75%, and 100% of examined BKT diluted with M9 buffer. All assays were replicated more than three times. Data are expressed as x±s, n=30. *P<0.05, **P<0.01, vs control group; △P<0.05, △△P<0.01, vs UV group."

Figure 3

Retrieval effects of BKT administration on egg number in uterus, brood size and generation time in heat-shock treated wild-type N2 nematodes HS, heat-shock. Approximately 30 L2-stage larvae grown on 20 ℃ were heat stressed for 2 h at 36 ℃ and afterwards further maintained at 20 ℃. Exposures were performed for 2 h on L4-larva stage nematodes with 25%, 50%, 75%, and 100% of examined BKT. All assays were replicated more than three times. Data are expressed as x±s, n=30. *P<0.05, **P<0.01, vs control group; ▲P<0.05, ▲▲P<0.01, vs HS group."

Figure 4

Retrieval effects of BKT administration on egg number in uterus, brood size and generation time in paraquat-treated wild-type N2 nematodes Approximately 30 L2-stage larvae were treated with 2 mmol/L paraquat solution for 2 h and survival at 20 ℃. Exposures were performed for 2 h on L4-larva stage nematodes with 25%, 50%, 75%, and 100% of examined BKT diluted with M9 buffer. All assays were replicated more than three times. Data are expressed as x±s, n=30. *P<0.05, **P<0.01, vs control group; □P<0.05, □□P<0.01, vs paraquat group."

Figure 5

Effects of BKT administration on CAT and SOD activities of UV-irradiated, heat-shock- and paraquat-treated wild-type N2 nematodes HS, heat-shock. Approximately 30 L2-stage larvae were irradiated on NGM plates without food at 20 J/(m2·min) at 20 ℃, or heat stressed for 2 h at 36 ℃, or treated with 2 mmol/L paraquat solution for 2 h and survival at 20 ℃. Exposures were performed for 2 h on L4-larva stage nematodes with 25%, 50%, 75%, and 100% of examined BKT diluted with M9 buffer. All assays were replicated more than three times. Data are expressed as x±s, n=30. *P<0.05, **P<0.01, vs control group; △P<0.05, △△P<0.01, vs UV group; ▲P<0.05, ▲▲P<0.01, vs HS group; □P<0.05, □□P<0.01, vs paraquat group."

Figure 6

Administration of BKT could retrieve the defects of egg number in uterus, brood size and generation time in mev-1 mutant nematodes N2, wild-type N2 nematodes. Exposures were performed for 2 h on L4-larva stage nematodes with 25%, 50%, 75%, and 100% of examined BKT diluted with M9 buffer. All assays were replicated more than three times. Data are expressed as x±s, n=30. *P<0.05, **P<0.01, vs control group; ■P<0.05, ■■P<0.01, vs mev-1 group."

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