观察大剂量维生素B₆对严重创伤后应激性肝损伤(HSI)的影响。

将HepG2细胞按维生素B₆终浓度0、0.5、1、2、4 mmol/L培养24 h后，使用CCK8试剂盒测定IC50。选用雄性SD大鼠按随机数字表法分为假模型组(n＝30)、假模型＋B₆组(n＝30)、创伤组(n＝30)、创伤＋B₆组(n＝30)，每个时间点(12、24、36、48和72 h)各6只。创伤组、创伤＋B₆组予腹壁损伤、双侧股骨骨折、单侧颅脑损伤、失血20%建立多发伤模型后予补液复苏。假模型组及假模型＋B₆组大鼠行股动脉穿刺抽血后，予补液复苏。复苏完成后12、24、36、48、72 h处死大鼠，留取血标本及肝脏组织。采用全自动生化仪检测丙氨酸氨基转氨酶(ALT)、天门冬氨酸氨基转移酶(AST)水平。苏木素-伊红(HE)染色、油红染色、透射电子显微镜观察大鼠肝脏组织病理学变化。

HepG2细胞的维生素B₆ IC50值为4 mmol/L。本研究维生素B₆使用剂量在安全范围内。与假模型组、假模型＋B₆组相比，大鼠严重创伤后12、24、36 h血清ALT、AST水平逐渐升高，48、72h血清ALT、AST水平逐渐下降。创伤＋B₆组的12、36、72h血清ALT水平显著均低于创伤组[(121.78±2.38)U/L比(177.41±6.00)U/L；(299.05±18.54)U/L比(447.03±22.02)U/L；(35.01±1.47)U/L比(48.32±4.79)U/L，P均<0.05]。创伤＋B₆组的12、24、36、48h的血清AST水平显著均低于创伤组[(601.52±17.27)U/L比(726.66±22.20)U/L；(619.44±45.05)U/L比(779.81±27.29)U/L；(672.36±16.50)U/L比(871.61±20.23)U/L；(133.26±19.11)U/L比(285.13±31.28 U/L)，P均<0.05]。HE染色可见大鼠肝脏损伤区在创伤后12 h集中于中央静脉与小叶间静脉的肝细胞。创伤后24、36h，肝损伤区域向中央静脉周围肝细胞和小叶间血管周围肝细胞进展。创伤后48、72h，肝损伤区域固定并逐渐缩小。各时间点创伤＋B₆组大鼠的肝细胞损伤范围及程度较创伤组均明显减轻。创伤后36 h油红染色显示创伤＋B₆组大鼠肝细胞内脂滴的数量及直径较创伤组明显下调。创伤后36 h透射电镜可见创伤＋B₆组肝组织较创伤组线粒体肿胀减轻，嵴恢复，可见杆状线粒体，脂滴、自噬前体、自噬小体、自噬溶酶体明显减少。

大剂量维生素B₆可通过减轻肝脏水肿变性及微泡性脂肪变性，减轻由严重创伤引起的HSI。创伤后36 h可能是创伤后HSI转归的重要时间点。

To assess the influence of a high-dose of vitamin B₆ on severe trauma-induced hepatic stress injury (HSI).

HepG2 cells were cultured with final concentrations of vitamin B₆ ranging from 0 to 4 mmol/L for 24 hours, and the IC50 was determined using the CCK8 assay. Male SD rats were selected and randomized into sham group (n=30), sham+ B₆ group (n=30), trauma group (n=30), and trauma+ B₆ group (n=30) using a random number table. At each time point (12、24、36、48 adn 72 h), there were 6 rats in ecah group. Severe multiple injuries were induced, including abdominal injury, bilateral femoral fractures, and unilateral cranial injury, followed by 20% blood loss and fluid resuscitation. Sham groups received arterial puncture and fluid resuscitation without severe injuries. Rats were euthanized at 12, 24, 36, 48, and 72 hours post-injury, and blood samples and liver tissues were collected. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured using an automatic biochemical analyzer. Pathological changes in liver tissues were observed through hematoxylin-eosin (HE) staining, oil red staining, and transmission electron microscope.

The IC50 value of vitamin B₆ for HepG2 cells was determined to be 4 mmol/L, confirming the safe dosage range used in this study. In comparison to sham groups, the serum ALT and AST levels in rats gradually increased at 12, 24, and 36 hours post-trauma and declined thereafter. The trauma+ B₆ group exhibited significantly lower ALT levels at 12, 36, and 72 hours [ (121.78±2.38) U/L vs. (177.41±6.00) U/L; (299.05±18.54) U/L vs. (447.03±22.02) U/L; (35.01±1.47) U/L vs. (48.32±4.79) U/L, all P<0.05]. Similarly, the trauma+ B₆ group showed significantly lower AST levels at 12, 24, 36, and 48 hours [ (601.52±17.27) U/L vs. (726.66±22.20) U/L; (619.44±45.05) U/L vs. (779.81±27.29) U/L; (672.36±16.50) U/L vs. (871.61±20.23) U/L; (133.26±19.11) U/L vs. (285.13±31.28) U/L, all P<0.05]. HE staining revealed that the injured area in rat livers was concentrated around the central vein and portal vein at 12 hours post-trauma, progressed to periportal hepatocytes and sinusoids at 24 and 36 hours, and became fixed and gradually reduced in size at 48 and 72 hours. The trauma+ B₆ group exhibited significantly reduced liver cell damage at all time points. Oil red staining at 36 hours post-trauma showed a significant downregulation of lipid droplet quantity and diameter in liver cells of the trauma+ B₆ group compared to the trauma group. Transmission electron microscopy at 36 hours post-trauma revealed reduced mitochondrial swelling, restored cristae, visible rod-shaped mitochondria, and a decrease in lipid droplets, autophagic precursors, autophagosomes, and autolysosomes in the trauma+ B₆ group compared to the trauma group.

High-dose vitamin B₆ can alleviate hepatic edema, degeneration, and microvesicular steatosis, reducing severe trauma-induced HSI. Additionally, the 36-hour time point post-trauma appears to be a critical juncture in the progression of HSI following severe trauma.