目的 探討甜菜堿對糖尿病足大鼠創(chuàng)面愈合及AMPK/SIRT1/FoxO1通路的影響。方法 SD大鼠隨機(jī)分為對照組、模型組、二甲雙胍（200 mg·kg^-1，ig給藥）組和甜菜堿低、高劑量（50、100 mg ·kg^-1，ip給藥）組，每組18只。除對照組外，其余組大鼠均構(gòu)建糖尿病足模型。檢測大鼠空腹血糖水平和創(chuàng)面愈合率；ELISA法檢測大鼠血清炎性因子白細(xì)胞介素-6（IL-6）、腫瘤壞死因子-α（TNF-α）、C反應(yīng)蛋白（CRP）水平；HE染色檢測大鼠創(chuàng)面皮膚組織病理學(xué)改變；免疫組化染色檢測大鼠創(chuàng)面皮膚組織中CD31蛋白陽性表達(dá)；試劑盒法檢測大鼠創(chuàng)面皮膚組織中氧化應(yīng)激指標(biāo)超氧化物歧化酶（SOD）活性、丙二醛（MDA）含量；Western blotting檢測大鼠創(chuàng)面皮膚組織中AMP依賴的蛋白激酶（AMPK）/沉默信息調(diào)節(jié)因子相關(guān)酶1（SIRT1）/叉頭框蛋白1（FoxO1）通路相關(guān)蛋白表達(dá)。結(jié)果 給藥結(jié)束后，與對照組比較，模型組大鼠空腹血糖、血清IL-6、TNF-α、CRP水平以及創(chuàng)面皮膚組織中MDA含量和p-FoxO1/FoxO1值顯著升高，創(chuàng)面愈合率和創(chuàng)面皮膚組織中CD31陽性表達(dá)率、SOD活性、p-AMPK/AMPK值和SIRT1蛋白表達(dá)水平顯著降低（P<0.05），且創(chuàng)面皮膚組織病理學(xué)損傷嚴(yán)重；與模型組比較，甜菜堿低、高劑量組和二甲雙胍組大鼠空腹血糖、血清IL-6、TNF-α、CRP水平以及創(chuàng)面皮膚組織中MDA含量和p-FoxO1/FoxO1值顯著降低，創(chuàng)面愈合率和創(chuàng)面皮膚組織中CD31陽性表達(dá)率、SOD活性、p-AMPK/AMPK值和SIRT1蛋白表達(dá)水平顯著升高（P<0.05），且創(chuàng)面皮膚組織病理學(xué)損傷均有不同程度改善，且甜菜堿高劑量組和二甲雙胍組上述指標(biāo)變化更為明顯。結(jié)論 甜菜堿可激活A(yù)MPK/SIRT1/FoxO1通路，減輕糖尿病足大鼠氧化應(yīng)激和炎癥反應(yīng)，促進(jìn)血管生成，加速創(chuàng)面愈合。

Objective To explore the effects of betaine on wound healing and AMPK/SIRT1/FoxO1 pathway in diabetes foot rats. Methods SD rats were randomly grouped into control group, model group, betaine (low and high) dose group (50, 100 mg·kg^-1) and metformin group (200 mg ·kg^-1), with 18 rats in each group. Except the control group, the rats in the other groups were constructed with diabetes foot models. The fasting blood glucose level and wound healing rate were measured. The levels of serum inflammatory factors (IL-6, TNF-α, CRP) were detected by ELISA. HE staining was applied to detect the histopathological changes of the wound skin. Immunohistochemical staining was applied to detect the positive expression of CD31 protein in the wound skin tissue of rats. The level of oxidative stress index (SOD activity, MDA content) in rat wound skin tissue was detected by commercial kit. Western blotting was applied to detect the expression of AMPK/SIRT1/FoxO1 pathway related protein in rat wound skin tissue.Results After administration, compared with the control group, the fasting blood glucose, serum IL-6, TNF-α, CRP levels, and the MDA content and p-FoxO1/FoxO1 ratio in the wound skin tissue of the model group were higher, the wound healing rate, the positive expression rate of CD31, the activity of SOD, the ratio of p-AMPK/AMPK and the expression level of SIRT1 protein in the wound skin tissue were lower (P< 0.05), the histopathological damage of the wound skin was serious. Compared with the model group, the fasting blood glucose, serum IL-6, TNF-α, CRP levels, and the MDA content and p-FoxO1/FoxO1 ratio in the wound skin tissue of rats in betaine (low and high) dose groups and metformin group were lower, the wound healing rate, the positive expression rate of CD31, the activity of SOD, the ratio of p-AMPK/AMPK and the expression level of SIRT1 protein in the wound skin tissue were higher (P< 0.05), in addition, the histopathological damage of the wound skin was improved to varying degrees, the changes of the above indexes were more obvious in betaine high-dose group and metformin group. Conclusion Betaine can activate AMPK/SIRT1/FoxO1 pathway, alleviate oxidative stress and inflammatory reaction, promote angiogenesis and accelerate wound healing in diabetes foot rats.

R965

北京市科技計(jì)劃項(xiàng)目（2021ZF073）