Unveiling the interplay between NSAID-induced dysbiosis and autoimmune liver disease in children: insights into the hidden gateway to autism spectrum disorders. Evidence from ex vivo, in vivo, and clinical studies

Doaa I Mohamed; Hebatallah H Abo Nahas; Asmaa M Elshaer; Dalia Alaa El-Din Aly El-Waseef; Omnyah A El-Kharashi; Soha M Y Mohamed; Yasmine Gamal Sabry; Riyad A Almaimani; Hussain A Almasmoum; Abdulmalik S Altamimi; Ibrahim Abdel Aziz Ibrahim; Samar Z Alshawwa; Mariusz Jaremko; Abdul-Hamid Emwas; Essa M Saied

doi:10.3389/fncel.2023.1268126

Unveiling the interplay between NSAID-induced dysbiosis and autoimmune liver disease in children: insights into the hidden gateway to autism spectrum disorders. Evidence from ex vivo, in vivo, and clinical studies

Front Cell Neurosci. 2023 Oct 31:17:1268126. doi: 10.3389/fncel.2023.1268126. eCollection 2023.

Authors

Doaa I Mohamed¹, Hebatallah H Abo Nahas², Asmaa M Elshaer¹, Dalia Alaa El-Din Aly El-Waseef³, Omnyah A El-Kharashi¹, Soha M Y Mohamed⁴, Yasmine Gamal Sabry⁴, Riyad A Almaimani⁵, Hussain A Almasmoum⁶, Abdulmalik S Altamimi⁷, Ibrahim Abdel Aziz Ibrahim⁸, Samar Z Alshawwa⁹, Mariusz Jaremko¹⁰, Abdul-Hamid Emwas¹¹, Essa M Saied^{12

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Affiliations

¹ Department of Clinical Pharmacology and Therapeutics, Faculty of Medicine, Ain Shams University, Cairo, Egypt.
² Physiology Department, Faculty of Science, Suez Canal University, Ismailia, Egypt.
³ Department of Histology and Cell Biology, Faculty of Medicine, Ain Shams University, Cairo, Egypt.
⁴ Physiology Department, Faculty of Medicine, Ain Shams University, Cairo, Egypt.
⁵ Department of Biochemistry, Faculty of Medicine, Umm Al-Qura University, Makkah, Saudi Arabia.
⁶ Department of Laboratory Medicine, Faculty of Applied Medical Sciences, Umm Al-Qura University, Makkah, Saudi Arabia.
⁷ Department of Pharmaceutical Chemistry, College of Pharmacy, Prince Sattam Bin Abdulaziz University, Alkharj, Saudi Arabia.
⁸ Department of Pharmacology and Toxicology, Faculty of Medicine, Umm Al-Qura University, Makkah, Saudi Arabia.
⁹ Department of Pharmaceutical Sciences, College of Pharmacy, Princess Nourah bint Abdulrahman University, Riyadh, Saudi Arabia.
¹⁰ Smart-Health Initiative and Red Sea Research Center, Division of Biological and Environmental Sciences and Engineering, King Abdullah University of Science and Technology, Thuwal, Saudi Arabia.
¹¹ Advanced Nanofabrication Imaging and Characterization Center, King Abdullah University of Science and Technology, Core Labs, Thuwal, Saudi Arabia.
¹² Chemistry Department, Faculty of Science, Suez Canal University, Ismailia, Egypt.
¹³ Institute for Chemistry, Humboldt Universität zu Berlin, Berlin, Germany.

Abstract

Autism spectrum disorders (ASD) represent a diverse group of neuropsychiatric conditions, and recent evidence has suggested a connection between ASD and microbial dysbiosis. Immune and gastrointestinal dysfunction are associated with dysbiosis, and there are indications that modulating the microbiota could improve ASD-related behaviors. Additionally, recent findings highlighted the significant impact of microbiota on the development of autoimmune liver diseases, and the occurrence of autoimmune liver disease in children with ASD is noteworthy. In the present study, we conducted both an in vivo study and a clinical study to explore the relationship between indomethacin-induced dysbiosis, autoimmune hepatitis (AIH), and the development of ASD. Our results revealed that indomethacin administration induced intestinal dysbiosis and bacterial translocation, confirmed by microbiological analysis showing positive bacterial translocation in blood cultures. Furthermore, indomethacin administration led to disturbed intestinal permeability, evidenced by the activation of the NLRP3 inflammasomes pathway and elevation of downstream biomarkers (TLR4, IL18, caspase 1). The histological analysis supported these findings, showing widened intestinal tight junctions, decreased mucosal thickness, inflammatory cell infiltrates, and collagen deposition. Additionally, the disturbance of intestinal permeability was associated with immune activation in liver tissue and the development of AIH, as indicated by altered liver function, elevated ASMA and ANA in serum, and histological markers of autoimmune hepatitis. These results indicate that NSAID-induced intestinal dysbiosis and AIH are robust triggers for ASD existence. These findings were further confirmed by conducting a clinical study that involved children with ASD, autoimmune hepatitis (AIH), and a history of NSAID intake. Children exposed to NSAIDs in early life and complicated by dysbiosis and AIH exhibited elevated serum levels of NLRP3, IL18, liver enzymes, ASMA, ANA, JAK1, and IL6. Further, the correlation analysis demonstrated a positive relationship between the measured parameters and the severity of ASD. Our findings suggest a potential link between NSAIDs, dysbiosis-induced AIH, and the development of ASD. The identified markers hold promise as indicators for early diagnosis and prognosis of ASD. This research highlights the importance of maintaining healthy gut microbiota and supports the necessity for further investigation into the role of dysbiosis and AIH in the etiology of ASD.

Keywords: NSAIDs; autism spectrum disorders; autoimmune hepatitis; clinical study; dysbiosis; gut microbiota; inflammasomes; prognostic biomarkers.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This research was funded by Faculty of Medicine, Ain Shams University, and Faculty of Science, Suez Canal University, Egypt. This research was also funded by Princess Nourah bint Abdulrahman University Researchers Supporting Project number (PNURSP2023R165), Princess Nourah bint Abdulrahman University, Riyadh, Saudi Arabia.