ABSTRACT
BACKGROUND: Posterior lumbar spine surgery is currently the main surgical approach for many surgical operations such as discectomy and spinal canal decompression. However, 10%-40% of patients with posterior lumbar spine surgery can have low back pain and related dysfunctions soon after surgery, which is related to excessive stretching or blunt injury to the paraspinal muscles such as the multifidus muscle during the operation. OBJECTIVE: To explore the effect of “Lumbar Three Needles” on the expression of oxidative stress factors, including malondialdehyde, reactive oxygen species, superoxide dismutase, glutathione peroxidase and phosphorylated protein kinase B, in rats with multifidus muscles injury. METHODS: Twenty-four male Sprague-Dawley rats were randomly divided into control group, model group and “Lumbar Three Needles” group, with 8 rats in each group. An animal model of multifidus muscles injury was made by intramuscular injection of 0.5% bupivacaine hydrochloride in the model and treatment groups in control and “Lumbar Three Needles” group, while normal saline was injected in the control group. No acupuncture intervention was given in the control group and model group, while Dachangshu, Shenshu and Weizhong acupoints were electroacupunctured in the “Lumbar Three Needles” group. Needles were then stimulated electrically using a Han's Acupoint Nerve Stimulator with density wave, frequency of 2 Hz/10 Hz and continuous current of 1 mA for 20 minutes, once daily for 7 days in total. The multifidus muscle was stained with hematoxylin-eosin staining to observe morphological changes, and kits were used to observe the expression of malondialdehyde, reactive oxygen species, superoxide dismutase, and glutathione peroxidase, while western blot was applied to observe the expression of phosphorylated protein kinase B. The study protocol was approved by the Animal Ethic Committee of Guangdong Second Traditional Chinese Medicine Hospital with an approval No. 048617. RESULTS AND CONCLUSION: Hematoxylin-eosin staining results indicated that the muscle fibers were partially fixed in the model group and there were still a lot of macrophages after 7 days of intervention. Compared with the model group, more newborn muscle fibers and fewer macrophages could be seen in the “Lumbar Three Needles” group. After intervention, the cross-sectional area of the multifidus muscles in the “Lumbar Three Needles” group was significantly bigger than that in the model group (P < 0.01). The expression levels of malondialdehyde and reactive oxygen species in the model group were significantly increased compared with the control group (P < 0.01) and “Lumbar Three Needles” group (P < 0.01). The expression of superoxide dismutase, glutathione peroxidase and phosphorylated protein kinase B in the “Lumbar Three Needles” group was significantly higher than that in the model group (P < 0.01 or P < 0.05). Therefore, “Lumbar Three Needles” can significantly decrease the oxidative stress level after multifidus muscle injury, which may be related to the improvement of phosphorylated protein kinase B.
ABSTRACT
ABSTRACT PURPOSE: We examined the effect of intracameral administration of cefuroxime on oxidative stress and endothelial apoptosis in rat corneal tissue. METHODS: In total, 30 rats were divided into 3 groups of 10 rats each (intracameral administration of cefuroxime 0.1 mg/0.01 mL (cefuroxime group); intracameral administration of balanced salt solution 0.01 mL (control group); or absence of intracameral injection (sham group). Corneal endothelial apoptosis was assessed by immunohistochemical analysis using caspase-3 and caspase-8. Total oxidant status, total antioxidant status, oxidative stress index, and paraoxonase and arylesterase levels were examined in corneal endothelial tissue and serum. RESULTS: Paraoxonase levels in the serum were significantly different between the sham and cefuroxime groups (p=0.027). A significant difference was also observed in total oxidant status levels between the cefuroxime and balanced salt solution groups (p=0.023). In addition, there were significant differences in total antioxidant status levels in corneal tissue between the cefuroxime and sham groups (p<0.001) and between the cefuroxime and balanced salt solution groups (p<0.001). Furthermore, significant differences were also observed in oxidative stress index levels between the cefuroxime and balanced salt solution groups (p=0.001) and between the cefuroxime and sham groups (p=0.026). According to the immunohistochemical staining results, a significant association with caspase-3 activity existed between the cefuroxime and balanced salt solution groups (p=0.007), while no significant difference was found with caspase-8 activity (p=0.541). Caspase-3 activity exhibited a significant relationship between the sham and balanced salt solution groups (p=0.018), but no relationship was found with caspase-8 activity (p=0.623). CONCLUSION: Immunohistochemical examination revealed that intracameral cefuroxime increased apoptosis when compared to the sham and balanced salt solution groups. Moreover, intracameral cefuroxime increased oxidative stress in the cornea and simultaneously induced apoptosis.
RESUMO OBJETIVO: Examinamos o efeito da administração intracameral da cefuroxima sobre o estresse oxidativo e a apoptose endotelial no tecido corneano de ratos. MÉTODOS: No total, 30 ratos foram divididos em 3 grupos de 10 ratos cada (administração intracameral de cefuroxima 0,1 mg/0,01 mL (grupo cefuroxima), administração intracameral de solução salina balanceada 0,01 mL (grupo controle) ou ausência de injeção intracameral (grupo sham)). A apoptose endotelial da córnea foi avaliada por análise imuno-histoquimica usando caspase-3 e -8. O status oxidante total, o status antioxidante total, o índice de estresse oxidativo e os níveis de a paraoxonase e arilesterase foram investigados no tecido endotelial da córnea e no soro. RESULTADOS: Os níveis de paraoxonase no soro foram significativamente diferentes entre os grupos sham e cefuroxima (p=0,027). Foi também observada uma diferença significativa nos níveis de estado oxidante total entre os grupos cefuroxima e solução salina balanceada (p=0,023). Além disso, houve diferenças significativas nos níveis de status antioxidante total no tecido da córnea entre os grupos cefuroxima e sham (p<0,001) e entre os grupos cefuroxima e solução salina balanceada (p<0,001). Diferenças significativas também foram observadas nos níveis do índice de estresse oxidativo entre os grupos cefuroxima e solução salina balanceada (p=0,001) e entre os grupos cefuroxima e sham (p=0,026). De acordo com os resultados de coloração imuno-histoquimica, houve associação significativa com a atividade da caspase-3 entre os grupos cefuroxima e solução salina balanceada (p=0,007), enquanto não houve diferença significativa com a atividade da caspase-8 (p=0,541). A atividade da caspase-3 exibiu uma relação significativa entre os grupos sham e solução salina balanceada (p=0,018), mas nenhuma relação foi encontrada com a atividade da caspase-8 (p=0,623). CONCLUSÃO: O exame imuno-histoquímico revelou que a cefuroxima intracameral aumentou a apoptose quando comparada com os grupos sham e solução salina balanceada. Além disso, a cefuroxima intracameral aumentou o estresse oxidativo na córnea e induziu simultaneamente a apoptose.