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<p>Microgravity alters gastrointestinal physiology, posing health risks during spaceflight. Krüppel-like factors (KLFs) are key transcriptional regulators of intestinal integrity, yet their behavior under microgravity in aged tissues remains unclear. Understanding these changes is vital for protecting gut health in aging individuals exposed to such environments. This study aimed to determine the effect of simulated microgravity on mRNA expression of KLFs in small intestine and colon cells and smooth muscles. This study employed a hindlimb unloading model to simulate microgravity conditions in laboratory-aged rats. A total of 15 male Sprague Dawley rats, aged 18–24 months, were divided into control (<i>n</i> = 7) and suspended (<i>n</i> = 8) groups, with suspension being induced for a period of two weeks. Following the intervention, small intestine and colon tissues were harvested for molecular analysis. Gene expression levels of KLFs (4, 5, 6 and 9) were quantitatively assessed using real-time reverse transcription polymerase chain reaction (RT-qPCR). Immunohistochemistry was employed to detect and quantify KLFs expression patterns. Significant morphological and molecular alterations were observed in intestinal tissues under simulated microgravity. Hindlimb suspension resulted in a pronounced reduction in small intestinal villus length (<i>P</i> &lt; 0.0001) and a marked decrease in Paneth cell count (<i>P</i> &lt; 0.0001). Gene expression analysis revealed tissue-specific modulation of Krüppel-like factors (KLFs). In the small intestine, KLF6 expression significantly declined (<i>P</i> = 0.047), while KLF9 levels increased (<i>P</i> = 0.035). In small intestinal smooth muscle, KLF5 was significantly downregulated (<i>P</i> = 0.009). Conversely, colonic smooth muscle exhibited significant upregulation of KLF4 (<i>P</i> = 0.032) and KLF5 (<i>P</i> &lt; 0.001), alongside a significant reduction in KLF9 (<i>P</i> = 0.021). This study highlighted the impact of simulated microgravity, via hindlimb suspension, on intestinal structure and KLF gene expression in aged rats. Villus length was significantly reduced, indicating impaired absorptive function, while Paneth cell depletion suggests weakened mucosal immunity. Although changes in crypt depth and muscle thickness were minimal, molecular analysis showed downregulation of KLF6 and upregulation of KLF9, pointing to disrupted epithelial homeostasis. KLF4 and KLF5 levels remained stable at the mRNA level.</p>

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Effect of Simulated Microgravity on Krüppel-Like Factor Expression in Gastrointestinal Cells of Aged Rats

  • 期刊论文

  • 发表日期：

    2026年3月23日

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  • Ahmed Al-Dwairi,
  • Tayseer Afifi,
  • Othman Al-Shboul,
  • Abdulsalam Bani Hamad,
  • Doa’a Al-u’datt

摘要

Microgravity alters gastrointestinal physiology, posing health risks during spaceflight. Krüppel-like factors (KLFs) are key transcriptional regulators of intestinal integrity, yet their behavior under microgravity in aged tissues remains unclear. Understanding these changes is vital for protecting gut health in aging individuals exposed to such environments. This study aimed to determine the effect of simulated microgravity on mRNA expression of KLFs in small intestine and colon cells and smooth muscles. This study employed a hindlimb unloading model to simulate microgravity conditions in laboratory-aged rats. A total of 15 male Sprague Dawley rats, aged 18–24 months, were divided into control (n = 7) and suspended (n = 8) groups, with suspension being induced for a period of two weeks. Following the intervention, small intestine and colon tissues were harvested for molecular analysis. Gene expression levels of KLFs (4, 5, 6 and 9) were quantitatively assessed using real-time reverse transcription polymerase chain reaction (RT-qPCR). Immunohistochemistry was employed to detect and quantify KLFs expression patterns. Significant morphological and molecular alterations were observed in intestinal tissues under simulated microgravity. Hindlimb suspension resulted in a pronounced reduction in small intestinal villus length (P < 0.0001) and a marked decrease in Paneth cell count (P < 0.0001). Gene expression analysis revealed tissue-specific modulation of Krüppel-like factors (KLFs). In the small intestine, KLF6 expression significantly declined (P = 0.047), while KLF9 levels increased (P = 0.035). In small intestinal smooth muscle, KLF5 was significantly downregulated (P = 0.009). Conversely, colonic smooth muscle exhibited significant upregulation of KLF4 (P = 0.032) and KLF5 (P < 0.001), alongside a significant reduction in KLF9 (P = 0.021). This study highlighted the impact of simulated microgravity, via hindlimb suspension, on intestinal structure and KLF gene expression in aged rats. Villus length was significantly reduced, indicating impaired absorptive function, while Paneth cell depletion suggests weakened mucosal immunity. Although changes in crypt depth and muscle thickness were minimal, molecular analysis showed downregulation of KLF6 and upregulation of KLF9, pointing to disrupted epithelial homeostasis. KLF4 and KLF5 levels remained stable at the mRNA level.