An extract of Rosaceae, Solanaceae and Zingiberaceae increases health span and mobility in Caenorhabditis elegans

Abstract

Background: Members of the Rosaceae, Solanaceae and Zingiberaceae families which include fruits such as cherries, tomatoes and ginger are known to have health promoting effects. There is growing interest in consuming these "functional foods" as a means to increase health and healthy ageing. However, many studies explore the effect of these foods in isolation, not as a blend of multiple functional foods.

Methods: In this study, an extract containing the dried berries, fruits, and roots of members of these families was prepared, which we called Bioact®180. The nematode Caenorhabditis elegans was used to evaluate the effects of Bioact®180 on lifespan and health endpoints, including muscle and mitochondria structure and locomotion.

Results: Exposure to the 1000 µg/mL of Bioact®180 extract, containing 4% total phenols, were healthier, as observed by an increase in mean lifespan with and small but significant increase in maximal lifespan. Nematodes exposed to Bioact®180 displayed better mobility in mid-life stages as well as enhanced mitochondrial morphology, which was more comparable to younger animals, suggesting that these worms are protected to some degree from sarcopenia.

Conclusions: Together, our findings reveal that Bioact®180, a blend of fruits and roots from Rosaceae, Solanaceae and Zingiberaceae family members has anti-aging effects. Bioact®180 promotes health and lifespan extension in C. elegans, corresponding to functional improvements in mobility.

Keywords: C. elegans; Health span; Mitochondria; Muscle; Plant extract; Sarcopenia.

Fig. 1

Lifespan is extended when worms are exposed to 1000 µg/mL to Bioact®180. (a) Survival curves of wild type (strain, N2) animals exposed to Bioact®180 supplemented NGM. The animals in control conditions (blue line; n = 276) have a similar curve to those exposed to the vehicle control (0.2% DMSO; red line; n = 192), 30 µg/mL Bioact®180 (green line; n = 178) and 100 µg/mL Bioact®180 (purple line; n = 203). Exposure to 1000 µg/mL Bioact®180 (orange line; n = 145) shifts the curve to the right, displaying an increase in health span as well as a slight increase in maximal lifespan. (b) The restricted mean lifespan is calculated from the survival curves [32, 33] and plotted here. Error bars represent the standard error of mean (s.e.m.) and statistical differences were compared to the control using the Fisher’s Exact test at 50% mortality, where */^#p < 0.05 and **/^##p < 0.001 and * compares data to the no compound control and # to the DMSO vehicle control. All data is further described in Table 1

Fig. 2

Bioact®180 does not affect the nuclear localisation of DAF-16::GFP. (a) Representative images of worms that have (i) cytoplasmic and (ii) nuclear DAF-16::GFP localisation. Scale bars are 100 µm. (b) Worms were placed onto Bioact®180 supplemented NGM as L1s and the localisation of DAF-16::GFP observed at the L4 stage. (c) From these, 5 random L4 animals were transferred to a fresh Bioact®180 supplemented NGM plate and allowed to lay eggs and then removed. When these progeny were L4, DAF-16::GFP localisation was recorded. Black bars indicate cytoplasmic localisation and white bars show DAF-16::GFP located in the nucleus where it is able to activate the insulin signalling pathway. Grey bars show an intermediate localisation, whereby DAF-16::GFP is found in both the nucleus and cytoplasm. The negative control was NGM, a 0.2% DMSO vehicle control was included together with a 2% DMSO positive control. Strain, TJ356; n = 100 for all conditions

Fig. 3

The mobility of animals exposed to Bioact®180 is maintained over time. Worms (strain, N2) were placed on no compound control (circles), 0.2% DMSO (squares) or 1000 µg/mL Bioact®180 (triangles) supplemented NGM as L4s (at T = 0). The number of head-to-tail body bends were assessed for each condition with the average and standard error of the mean shown. On day 4, worms in control conditions (open circles; n = 18) had the same number of body bends per minute, BBPM, as those exposed to DMSO (open squares; n = 19) or Bioact®180 (open triangles; n = 15). When the worms are 8 days of age, the number of BBPM significantly decreases when worms are exposed to control conditions (closed circles; n = 18) and the DMSO vehicle control (closed squares; n = 18). In contrast, worms on 1000 µg/mL Bioact®180 (closed tringles; n = 10) show no change in BBPM, suggesting that the compound is providing some health benefit. p-value obtained by the 2-tailed 2-sample t-test, where *p < 0.05 and **p < 0.001. If no asterisks, there is no significant difference

Fig. 4

Muscle morphology is unchanged in Bioact®180 exposed worms compared to wild type. Worms (strain, RW1596) were placed on no compound control, 0.2% DMSO or 1000 µg/mL Bioact®180 supplemented NGM as L4s (at T = 0). The muscle morphology was observed via the myo-3p::GFP reporter for each condition on days 4, 8 and 15 and assessed as being linear or fragmented. (a) Representative image of a worm displaying (i) linear or (ii) fragmented muscle fibres. Scale bars, 100 µm. (b) Graph to show the percentage of worms in each category at the different time points. Black bars indicate linear muscle morphology while the grey bars are for those fibres which are fragmented. Data was from 4 independent biological replicates with more than 85 worms assessed on day 4 (control n = 88; DMSO n = 85; Bioact®180 n = 85) and more than 95 worms on day 8 (control n = 110; DMSO n = 95; Bioact®180 n = 93), and around 20 worms on day 15 (control n = 28; DMSO n = 19; Bioact®180 n = 24)

Fig. 5

Mitochondrial morphology of worms exposed to Bioact®180 have less degradation on day 8 of life. Worms (strain, SD1347) were placed on no compound control, 0.2% DMSO or 1000 µg/mL Bioact®180 supplemented NGM as L4s (at T = 0). The mitochondrial network of worms was assessed for linear, intermediate or fragmented morphology on day 4 and 8. (a) Representative image of a worm displaying a (i) linear or (ii) fragmented mitochondrial network. Scale bars, 100 µm. Graphs to show the mitochondrial morphology on (b) day 4 and (c) day 8. The classifications are as follows: linear (black bars) where the mitochondrial network has even linear lines; fragmented (white bars) is where the network has reduced significantly and is mostly broken; an intermediate classification (grey bars) is where the network is uneven and starting to fragment. The mitochondrial morphology was assessed over 2 independent experiments, with a total of n = 40 for day 4 control, day 4 Bioact®180 and day 8 Bioact®180; n = 38 for the DMSO control at day 4 and 8; n = 41 for control worms on day 8