Medicinal Plants for the Treatment of Mental Diseases in Pregnancy: An In Vitro Safety Assessment

Pregnancy is a critical period for medical care, during which the well-being of woman and fetus must be considered. This is particularly relevant in managing non-psychotic mental disorders since treatment with central nervous system-active drugs and untreated NMDs may have negative effects. Some well-known herbal preparations (phytopharmaceuticals), including St. Johnʼs wort, California poppy, valerian, lavender, and hops, possess antidepressant, sedative, anxiolytic, or antidepressant properties and could be used to treat mental diseases such as depression, restlessness, and anxiety in pregnancy. Our goal was to assess their safety in vitro, focusing on cytotoxicity, induction of apoptosis, genotoxicity, and effects on metabolic properties and differentiation in cells widely used as a placental cell model (BeWo b30 placenta choriocarcinoma cells). The lavender essential oil was inconspicuous in all experiments and showed no detrimental effects. At low-to-high concentrations, no extract markedly affected the chosen safety parameters. At an artificially high concentration of 100 µg/mL, extracts from St. Johnʼs wort, California poppy, valerian, and hops had minimal cytotoxic effects. None of the extracts resulted in genotoxic effects or altered glucose consumption or lactate production, nor did they induce or inhibit BeWo b30 cell differentiation. This study suggests that all tested preparations from St. Johnʼs wort, California poppy, valerian, lavender, and hops, in concentrations up to 30 µg/mL, do not possess any cytotoxic or genotoxic potential and do not compromise placental cell viability, metabolic activity, and differentiation. Empirical and clinical studies during pregnancy are needed to support these in vitro data

Intestinal permeability and gut microbiota interactions of pharmacologically active compounds in valerian and St. John’s wort

Phytomedicines such as valerian and St. John's wort are widely used for the treatment of sleeping disorders, anxiety and mild depression. They are perceived as safe alternatives to synthetic drugs, but limited information is available on the intestinal absorption and interaction with human intestinal microbiota of pharmacologically relevant constituents valerenic acid in valerian, and hyperforin and hypericin in St. John's wort. The intestinal permeability of these compounds and the antidepressant and anxiolytic drugs citalopram and diazepam was investigated in the Caco-2 cell model with bidirectional transport experiments. In addition, interaction of compounds and herbal extracts with intestinal microbiota was evaluated in artificial human gut microbiota. Microbiota-mediated metabolisation of compounds was assessed, and bacterial viability and short-chain fatty acids (SCFA) production were measured in the presence of compounds or herbal extracts. Valerenic acid and hyperforin were highly permeable in Caco-2 cell monolayers. Hypericin showed low-to-moderate permeability. An active transport process was potentially involved in the transfer of valerenic acid. Hyperforin and hypericin were mainly transported through passive transcellular diffusion. All compounds were not metabolized over 24 h in the artificial gut microbiota. Microbial SCFA production and bacterial viability was not substantially impaired nor promoted by exposure to the compounds or herbal extracts

Placental Passage of Protopine in an Ex Vivo Human Perfusion System

The placental passage of protopine was investigated with a human ex vivo placental perfusion model. The model was first validated with diazepam and citalopram, 2 compounds known to cross the placental barrier, and antipyrine as a positive control. All compounds were quantified by partially validated U(H)PLC-MS/MS bioanalytical methods. Protopine was transferred from the maternal to the fetal circuit, with a steady-state reached after 90 min. The study compound did not affect placental viability or functionality, as glucose consumption, lactate production, and beta-human chorionic gonadotropin, and leptin release remained constant. Histopathological evaluation of all placental specimens showed unremarkable, age-appropriate parenchymal maturation with no pathologic findings

Transplacental passage of hyperforin, hypericin, and valerenic acid

Safe medications for mild mental diseases in pregnancy are needed. Phytomedicines from St. John’s wort and valerian are valid candidates, but safety data in pregnancy are lacking. The transplacental transport of hyperforin and hypericin (from St. John’s wort), and valerenic acid (from valerian) was evaluated using the ex vivo cotyledon perfusion model (4 h perfusions, term placentae) and, in part, the in vitro Transwell assay with BeWo b30 cells. Antipyrine was used for comparison in both models. U(H)PLC-MS/MS bioanalytical methods were developed to quantify the compounds. Perfusion data obtained with term placentae showed that only minor amounts of hyperforin passed into the fetal circuit, while hypericin did not cross the placental barrier and valerenic acid equilibrated between the maternal and fetal compartments. None of the investigated compounds affected metabolic, functional, and histopathological parameters of the placenta during the perfusion experiments. Data from the Transwell model suggested that valerenic acid does not cross the placental cell layer. Taken together, our data suggest that throughout the pregnancy the potential fetal exposure to hypericin and hyperforin – but not to valerenic acid – is likely to be minimal

Transplacental passage of hyperforin, hypericin, and valerenic acid

Safe medications for mild mental diseases in pregnancy are needed. Phytomedicines from St. John’s wort and valerian are valid candidates, but safety data in pregnancy are lacking. The transplacental transport of hyperforin and hypericin (from St. John’s wort), and valerenic acid (from valerian) was evaluated using the ex vivo cotyledon perfusion model (4 h perfusions, term placentae) and, in part, the in vitro Transwell assay with BeWo b30 cells. Antipyrine was used for comparison in both models. U(H)PLC-MS/MS bioanalytical methods were developed to quantify the compounds. Perfusion data obtained with term placentae showed that only minor amounts of hyperforin passed into the fetal circuit, while hypericin did not cross the placental barrier and valerenic acid equilibrated between the maternal and fetal compartments. None of the investigated compounds affected metabolic, functional, and histopathological parameters of the placenta during the perfusion experiments. Data from the Transwell model suggested that valerenic acid does not cross the placental cell layer. Taken together, our data suggest that throughout the pregnancy the potential fetal exposure to hypericin and hyperforin – but not to valerenic acid – is likely to be minimal

Safety assessment of herbal drugs as alternatives to treat non-psychotic mental diseases during pregnancy

Pregnancy is an extraordinary state for the expecting mother. Her body goes through many changes to bring forth new life. Amidst the joy of bearing a child, up to 20% of pregnant women will suffer from depression. Other ailments are anxiety, restlessness, stress, and sleeping issues, which are categorized as non-psychotic mental diseases (NMDs). In order to prevent further discomfort for the expecting mothers and negative birth outcomes like preterm birth or preeclampsia, medications are prescribed to mitigate the symptoms. Typical treatments are selective serotonin reuptake inhibitors (e.g., citalopram) and benzodiazepines (e.g., diazepam) to treat mild depression and anxiety, respectively. Those synthetic drugs are for instance known to cross the placental barrier, which might be one of many factors having physicians and pregnant women discuss possible alternatives. Alternatives to treat NMDs are valerian, lavender, hops, St. John’s Wort (SJW), and California poppy. Treatment with herbal medication is as old as the history of medicine itself and remedies of natural origin have ever since been considered trustworthy. This notion is also reflected in the different handling of synthetic drugs versus herbal ones. The regulations worldwide differ. In some countries herbal medications are categorized as dietary supplements. Although the Committee on Herbal Medicinal Products has regulations in place for herbal medications; data on those products are often missing and most herbal medication are simply labelled as not recommended during pregnancy and lactation. The high trust in herbal medications and the lack of safety data at the same time, potentially expose pregnant women and their unborn child to unknown risks. To assess the safety of valerian, hops, SJW, and California poppy we first established bioanalytical methods for the compounds attributed to the therapeutic effect, namely valerenic acid, humulone, hyperforin, hypericin, and protopine. Lavender and its volatile constituent linalool have been excluded from the following studies. In a first step the bioanalytical methods allowed determination of foetal exposure to those single compounds in an ex vivo placental perfusion model. We could show that protopine and valerenic acid crossed the placental barrier, while hyperforin only transferred to a small extent into the foetal circuit. Hypericin on the other hand, accumulated in the placenta. To make an informed safety assessment, knowledge of potential metabolites is also needed. We therefore investigated phase I and phase II metabolism for valerenic acid, protopine, and humulone in human liver microsomes. For valerenic acid, we were able to identify five dehydrovalerenic acid isomers, valerenic acid glucuronide, and oxoglucuronide. Protopine was metabolised to dimethylcryptopine, and to a metabolite formed by catechol-O-methylation and a third metabolite formed by ketone reduction. Finally, for humulone we could identifiy eleven hydroxyhumulone isomers, three dihydroxyhumulone isomers, and a metabolite formed by aliphatic hhydroxylation and ketone reduction. Those metabolites have been described in human liver microsomes for the first time. The third part of the project was dedicated to screening for potential mitochondria targeted liver toxicity in HepG2 and HepaRG cells. Hypericin was, in term of toxic effects, the most promising candidate. Hypericin turned out to be a potent mitochondria toxicant. It showed inhibition of complexes I, III, and IV in the electron transport chain and a decrease in mitochondrial ATP production, which leads to mitochondrial reactive oxygen species accumulation. Nrf2 was activated, which eventually leads to necrosis. In conclusion we could show that the foetus is directly exposed to valerenic acid and protopine, while exposure to hyperforin is minimal. Further studies to determine the potential harm for the unborn are still needed. An investigation for possible influences of the accumulation of hypericin in the placenta (change in the viability of the placenta and therefore consequences for the foetus) also needs to be carried out further since we now know the toxicant potential of hypericin. Additionally, a study of the identified metabolites and their interactions with the body are necessary. Finally, an assessment of the full extracts in vitro and in vivo is required since those single compounds are rarely consumed alone, but as part of complex matrix in the herbal medication. With the obtained results we cannot conclude that the investigated compounds or the full extracts are safe

Intestinal permeability and gut microbiota interactions of pharmacologically active compounds in valerian and St. John’s wort

Phytomedicines such as valerian and St. John’s wort are widely used for the treatment of sleeping disorders, anxiety and mild depression. They are perceived as safe alternatives to synthetic drugs, but limited information is available on the intestinal absorption and interaction with human intestinal microbiota of pharmacologically relevant constituents valerenic acid in valerian, and hyperforin and hypericin in St. John’s wort. The intestinal permeability of these compounds and the antidepressant and anxiolytic drugs citalopram and diazepam was investigated in the Caco-2 cell model with bidirectional transport experiments. In addition, interaction of compounds and herbal extracts with intestinal microbiota was evaluated in artificial human gut microbiota. Microbiota-mediated metabolisation of compounds was assessed, and bacterial viability and short-chain fatty acids (SCFA) production were measured in the presence of compounds or herbal extracts. Valerenic acid and hyperforin were highly permeable in Caco-2 cell monolayers. Hypericin showed low-to-moderate permeability. An active transport process was potentially involved in the transfer of valerenic acid. Hyperforin and hypericin were mainly transported through passive transcellular diffusion. All compounds were not metabolized over 24 h in the artificial gut microbiota. Microbial SCFA production and bacterial viability was not substantially impaired nor promoted by exposure to the compounds or herbal extracts

Retraction: Placental Passage of Humulone and Protopine in an Ex Vivo Human Perfusion System

The authors have retracted this paper after they discovered that one of the compounds commercially obtained was in fact another substance. This unfortunate situation was beyond the control of the authors who regret any confusion that may have been caused by the published results

DataSheet1_Transplacental passage of hyperforin, hypericin, and valerenic acid.PDF

Safe medications for mild mental diseases in pregnancy are needed. Phytomedicines from St. John’s wort and valerian are valid candidates, but safety data in pregnancy are lacking. The transplacental transport of hyperforin and hypericin (from St. John’s wort), and valerenic acid (from valerian) was evaluated using the ex vivo cotyledon perfusion model (4 h perfusions, term placentae) and, in part, the in vitro Transwell assay with BeWo b30 cells. Antipyrine was used for comparison in both models. U(H)PLC-MS/MS bioanalytical methods were developed to quantify the compounds. Perfusion data obtained with term placentae showed that only minor amounts of hyperforin passed into the fetal circuit, while hypericin did not cross the placental barrier and valerenic acid equilibrated between the maternal and fetal compartments. None of the investigated compounds affected metabolic, functional, and histopathological parameters of the placenta during the perfusion experiments. Data from the Transwell model suggested that valerenic acid does not cross the placental cell layer. Taken together, our data suggest that throughout the pregnancy the potential fetal exposure to hypericin and hyperforin – but not to valerenic acid – is likely to be minimal.</p