Assessing the therapeutic potential and safety of traditional anti-obesity herbal blends in Palestine

Assessing the therapeutic potential and safety of traditional anti-obesity herbal blends in Palestine

The increasing use of traditional medicinal herbs has raised concerns about potential side effects and misapplication. Herbal products have been linked to a variety of negative side effects, including life-threatening conditions and serious injuries. Some of the difficulties in monitoring the safety of herbal remedies stem from the lack of mandatory safety evaluations before marketing, as well as the lack of quality standards regulations, and effective manufacturing practices in many countries44,45. Therefore, our study focused on the biological evaluation of two traditional herbal remedies containing eight herbal plants commonly used in traditional Palestinian medicine in the West Bank of Palestine and evaluate their anti-obesity, antidiabetic, antifibrotic, antioxidant, and anti-cancer effects.

The anti-oxidant activities of the evaluated Herbal blends were evaluated using the DPPH assay, and the results are displayed in Fig. 1. Herbal Blend 1 exhibited an IC50 value of 68.16 ± 2.45 µg/mL, while Herbal Blend 2 showed an IC50 value of 33.97 ± 1.14 µg/mL, compared to a positive control (Trolox) with an IC50 of 7.72 ± 1.05 µg/mL, as indicated in Table 2.

Figure 1
figure 1

DPPH free radicals scavenging property of two herbal blends and Trolox.

Table 2 The IC50 Values in µg/mL on each biological target of both Herbal blends 1 and 2 in comparison with the positive control.

It was found that Herbal Blend 2 had a more potent antioxidant activity compared to Herbal Blend 1. The higher antioxidant activity of Herbal Blend 2 can be attributed to the presence of C. longa, A. sativum, and P. nigrum. In fact, C. longa is known to contain curcumin, which has been extensively studied for its potential antioxidant properties46. Similarly, A. sativum contains sulfur-containing compounds that also possess antioxidant properties. A study published in the Journal of Agricultural and Food Chemistry showed that garlic extracts have high antioxidant activity, as measured by the ORAC (Oxygen Radical Absorbance Capacity) assay47. Besides, an investigation established by Nahak and Sahu found that P. nigrum fruits exhibited remarkable DPPH free radicals scavenging ability at different concentrations48. Moreover, P. nigrum enhances the bioavailability of curcumin, thereby increasing the antioxidant potential of Herbal Blend 2. Although Herbal Blend 1 ingredients may also have some antioxidant potential, they may not be as potent as the combination of C. longa, A. sativum, and P. nigrum found in Herbal Blend 2.

The percentage of inhibition was measured for the Herbal blends at different concentrations, and the results are illustrated in Fig. 1. Herbal Blend 2 showed a significantly higher percentage of inhibition than Herbal Blend 1, and in some concentrations, it even surpassed the control (Trolox). For instance, at a concentration of 50 µg/mL, Herbal Blend 2 exhibited a percentage of inhibition of 68%, while Trolox showed 61%.

Herbal remedies have been used for centuries in traditional medicine for their anticancer and cytotoxic effects. These effects are attributed to the presence of various active compounds in plants, such as triterpene saponins, flavonoids, and phenolic compounds, which have been shown to inhibit cancer cell growth, induce apoptosis, and cause DNA damage49.

The anti-cancer activities of the tested Herbal blends were evaluated using the MTS assay, and the results showed that Herbal Blend 2 had potent anticancer activities against liver cancer and breast cancer compared to Herbal Blend 1, which also exhibited anticancer activities by inducing apoptosis in liver and breast cancer cells. The percentage of cell viability was measured at different concentrations for the Herbal blends, as shown in Fig. 2.

Figure 2
figure 2

Cytotoxic activity of Two herbal blends, DMSO and DOX on Hep3B, MCF-7, and HeLa cancer cells.

Based on the results of the biological evaluation, it was found that Herbal Blend 2 had a more potent anticancer activity compared to Herbal Blend 1. One possible reason for this difference in activity could be attributed to the presence of C. longa and A. sativum in Herbal Blend 2. C. longa contain curcumin, which has been extensively studied for its anticancer properties, Curcumin, employed in Ayurvedic medicine for its anti-inflammatory properties, has been reported to synergistically inhibit tumor cell growth and induce apoptosis50. Similarly, A. sativum contains organosulfur compounds including allicin, which have also been shown to possess anticancer properties51. Piper nigrum, which is present in both Herbal blends, is known to enhance the bioavailability of curcumin, potentially increasing its activity52. Additionally, previous studies explored the anticancer activity of P. nigrum fruits in both in vitro and in vivo breast cancer models which showed that the treatments with the plant extracts induced intracellular oxidative stress, which was considered the main component responsible for its cytotoxic effects in cancer cells53. In contrast, While these ingredients in Herbal Blend 1 may also possess some medicinal properties, they may not have the same level of anticancer activity as C. longa. and A. sativum found in Herbal Blend 2.

For Hep3B cancer cell lines, the IC50 of Herbal Blend 1 was 148.37 ± 2.45 µg/mL, while the IC50 of Herbal Blend 2 was 71.74 ± 1.77 µg/mL, compared to a positive control (DOX) with an IC50 of 1.21 ± 1.0 µg/mL, as shown in Table 2. Similarly, for MCF-7 cells, the IC50 of Herbal Blend 1 was 127.10 ± 2.08 µg/mL, while the IC50 of Herbal Blend 2 was 52.53 ± 0.78 µg/mL, compared to a positive control (DOX) with an IC50 of less than 1, as shown in Table 2. In addition, for HeLa cells, the IC50 of Herbal Blend 1 was 258.06 ± 1.07 µg/mL, while the IC50 of Herbal Blend 2 was 164.89 µg/mL, compared to a positive control (DOX) with an IC50 of 1.55 µg/mL, as presented in Table 2. These results indicate that Herbal Blend 2 has a stronger anti-cancer activity against liver cancer and breast cancer cells compared to Herbal Blend 1.

C. verum commonly known as Cinnamon, exhibits anti-cancer, antioxidant, anti-lipid, and anti-fibrotic properties17,54 It demonstrates anticancer effects against various cancer cell types, including breast cancer, through apoptosis induction and angiogenesis suppression. This extends to its efficacy against lung and colon cancer cells55,56. C. longa, or Turmeric, is recognized for its potent anti-cancer properties, effectively inhibiting the growth of various cancer cells such as those in breast, prostate, and colon cancers57,58,59. Additionally, C. longa displays significant antioxidant activity, contributing to the prevention of disorders associated with oxidative stress, including cardiovascular diseases and cancer57,60. The herb’s anti-lipid properties are notable, demonstrated by its ability to reduce blood glucose and lipid levels in diabetic mice. Furthermore, Curcuma longa showcases anti-fibrotic properties, suggesting potential benefits in preventing and treating fibrotic conditions like liver fibrosis20,21,61.

The anti-fibrotic activity of the tested Herbal blends was also evaluated, and it was found that Herbal Blend 2 exhibited more potent anti-fibrotic activity compared to Herbal Blend 1. The IC50 values for Herbal Blend 2 and Herbal Blend 1 in the anti-cancer assay were 24.36 ± 0.54 and 83.25 ± 2.45 µg/mL, respectively, while the control drug DOX showed an IC50 of < 1, as shown in Table 2. These results suggest that the tested Herbal blends may have potential therapeutic applications not only for fibrotic conditions but also for cancer treatment due to their multi-faceted activities. The findings of this study provide a basis for further research and development of these Herbal blends as potential anti-fibrotic, anti-cancer, and antioxidant agents.

Herbal Blend 2 had a more potent antifibrotic activity compared to Herbal Blend 1. The higher antifibrotic activity of Herbal Blend 2 could be due to the presence of C. longa and A. sativum in the Herbal blend. Curcumin, present in C. longa has been reported to have antifibrotic properties by inhibiting fibrosis-promoting factors62. Additionally, A. sativum contains sulfur-containing compounds that possess antifibrotic properties63. Therefore, the combination of C. longa and A. sativum in Herbal Blend 2 may be responsible for its higher antifibrotic activity.

When considering all of these activities together, it is evident that Herbal Blend 2 has the potential to provide multiple benefits for individuals with liver cancer. The anti-cancer activity of Herbal Blend 2 against Hep3B cells, combined with its anti-fibrotic activity, may help to prevent the progression of liver fibrosis, a common complication of liver cancer. Furthermore, the antioxidant activity of Herbal Blend 2 may help to protect liver cells from oxidative stress, which is known to contribute to the development and progression of liver cancer. Therefore, the multi-faceted benefits of Herbal Blend 2 make it a promising candidate for further investigation and development as a potential therapeutic agent for liver cancer.

Figure 3 displays the percentage of LX-2 cells inhibition at various concentrations for both Herbal Blend 1 and Herbal Blend 2, demonstrating promising results. The inhibition percentages for the Herbal blends were compared with those of 5FU and DOX. At a concentration of 80 µg/mL, Herbal Blend 2 exhibited an inhibition percentage of 68%, while Herbal Blend 1 displayed a percentage of 50%. In contrast, the inhibition percentages of 5-Fu and DOX were 72% and 75%, respectively. Moreover, at a concentration of 250 µg/mL, Herbal Blend 2 demonstrated a percentage of 77%, which was almost equivalent to the percentage of Herbal Blend 1 (77%). However, the LX-2 cells inhibition percentages of 5-Fu and DOX were 95% and 75%, respectively. These results indicate that both Herbal blends have significant inhibitory effects on LX-2 cancer cells and suggest that they could be effective alternatives to traditional chemotherapy drugs.

Figure 3
figure 3

Antifibrotic effect of two herbal blends, 5-FU and DOX on LX-2 cell lines.

The anti-diabetic and anti-obesity potential of the tested Herbal blends was evaluated by examining their effects on porcine pancreatic α-amylase and lipase enzymes. The results showed that Herbal Blend 2 exhibited more potent anti-amylase activity compared to Herbal Blend 1, with IC50 doses of 243.73 ± 1.57 and 468.98 ± 2.44 µg/mL, respectively. In contrast, the control drug Acarbose displayed an IC50 value of 6.42 ± 1.02 µg/mL, indicating that both Herbal blends have moderate anti-amylase effects. However, there was a negligible effect on lipase activity for both Herbal blends, as Herbal Blend 2 and Herbal Blend 1 exhibited IC50 values of 1358.39 ± 2.04 and 1466.85 ± 3.54 µg/mL, respectively, whereas the control drug Orlistat had an IC50 value of 5.44 µg/mL, as shown in Table 2. These findings suggest that the evaluated Herbal blends could be used as natural treatments for diabetes and obesity due to their significant anti-amylase activity with caution because of their potent activities on various cell lines.

Herbal Blend 2 had a more potent anti-amylase activity compared to Herbal Blend 1. The higher anti-amylase activity of Herbal Blend 2 could be due to the presence of C. verum and P. nigrum in the Herbal blend. Both of these ingredients are known to possess anti-amylase properties. For example, cinnamon extract has been reported to inhibit α-amylase, an enzyme involved in carbohydrate digestion, and reduce the postprandial glucose response64. Additionally, P. nigrum contains piperine, which has been shown to inhibit α-amylase and reduce starch digestion65. Therefore, the combination of C. verum and P. nigrum in Herbal Blend 2 may be responsible for its higher anti-amylase activity.

Herbal Blend 1 and Herbal Blend 2, revealed that they had a negligible effect on lipase enzyme activity. The reason for this could be due to the absence of specific ingredients in both Herbal blends that are known to have lipase-inhibitory properties. For example, some studies have shown that polyphenols, such as epigallocatechin-3-gallate (EGCG) found in green tea (C. sinensis) and curcuminoids found in turmeric (C. longa ), can inhibit pancreatic lipase activity66. However, the levels of these polyphenols in Herbal Blend 1 and Herbal Blend 2 may not have been sufficient to have a significant impact on lipase enzyme activity.

Figure 4 displays the percentage of inhibition at different concentrations of the tested Herbal blends on α-amylase, and the results were promising for both Herbal blends compared to the control Acarbose. Herbal Blend 2 demonstrated a percentage of inhibition of 59% at a concentration of 510 µg/mL, while Herbal Blend 1 showed 44% inhibition at the same concentration. In comparison, the percentage of inhibition for Acarbose was 64%. At a concentration of 730 µg/mL, both Herbal blends showed similar percentages of inhibition, as Herbal Blend 2 demonstrating a 60% inhibition and Herbal Blend 1 showing 58% of inhibition, while the percentage of inhibition for Acarbose was 67%. These results indicate that both Herbal blends have anti-diabetic activity through their inhibition of α-amylase, and Herbal Blend 2 was found to have slightly more potent activity compared to Herbal Blend 1.

Figure 4
figure 4

Anti-amylase effect of Two herbal blends and Acarbose.

Figure 5 presents the percentage of inhibition at different concentrations of the two tested Herbal blends against lipase. The results showed negligible effects for both Herbal blends compared to Orlistat. At a concentration of 510 µg/mL, Herbal Blend 2 had almost the same percentage of inhibition as Herbal Blend 1, with Herbal Blend 2 showing a percentage of 35% and Herbal Blend 1 showing 37%, while Orlistat exhibited a percentage of 97%. Similarly, at a concentration of 730 µg/mL, Herbal Blend 2 showed a percentage of 44%, and Herbal Blend 1 showed 42%, while Orlistat exhibited a percentage of 99%. These results suggest that the two tested Herbal blends have a negligible effect on lipase activity compared to Orlistat.

Figure 5
figure 5

Anti-lipase effect of two Herbal blends and Orlistat.

Further standardization of the active components in the investigated herbal blends, along with in vivo and preclinical research, is necessary to validate the findings of our study and develop appropriate pharmaceutical dosages from these traditional blends to enhance community healthcare.

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