附子对高糖刺激下施万细胞中Krox20-Oct6信号通路及其调控的髓鞘蛋白的影响

目的观察附子对高糖培养下施万细胞中Krox20-Oct6途径及其调控的髓鞘蛋白的影响,以揭示附子治疗糖尿病周围神经病变的机制。方法将施万细胞分为以下6组:(1)正常组(低浓度葡萄糖);(2)对照组(甘露醇);(3)模型组(高浓度葡萄糖);(4)附子水提物高、中、低剂量组(10μg/mL,1.0μg/mL,0.1μg/mL)。常规培养3天后,采用实时荧光定量PCR法检测各组细胞Oct6、Krox20、髓鞘碱性蛋白和髓鞘蛋白Z mRNA的表达水平,采用Western blot法检测各组细胞的Oct6和Krox20蛋白的表达水平,采用免疫荧光法检测各组细胞的髓鞘碱性蛋白和髓鞘蛋白Z蛋白的表达水平。结果 PCR结果显示,与正常组相比,模型组中Oct6、Krox20、髓鞘碱性蛋白和髓鞘蛋白Z m RNA表达水平下降;与模型组比,附子水提物各剂量组中Oct6、Krox20、髓鞘碱性蛋白和髓鞘蛋白Z m RNA表达水平上升。Western blot结果证明,与正常组相比,模型组中Oct6蛋白、Krox20蛋白表达水平下降;与模型组比,附子水提物各剂量组中Oct6蛋白、Krox20蛋白表达水平上升。免疫荧光结果提示,与正常组相比,模型组中髓鞘碱性蛋白蛋白及髓鞘蛋白Z蛋白表达水平下降;与模型组比,附子水提物各剂量组中髓鞘碱性蛋白蛋白及髓鞘蛋白Z蛋白表达水平上升。结论高浓度葡萄糖通过抑制Krox20-Oct6途径使施万细胞髓鞘蛋白生成能力下降,而附子可能通过Krox20-Oct6途径促进髓鞘蛋白的形成,从而改善糖尿病周围神经病变。

OCT6 inhibits differentiation of porcine-induced pluripotent stem cells through MAPK and PI3K signaling regulation

As a transcription factor of the Pit-Oct-Unc(POU)domain family,octamer-binding transcription factor 6(OCT6)participates in various aspects of stem cell development and differentiation.At present,however,its role in porcine-induced pluripotent stem cells(piPSCs)remains unclear.Here,we explored the function of OCT6 in piPSCs.We found that piPSCs overexpressing OCT6 maintained colony morphology and pluripotency under differentiation conditions,with a similar gene expression pattern to that of non-differentiated piPSCs.Functional analysis revealed that OCT6 attenuated the adverse effects of extracellular signal-regulated kinase(ERK)signaling pathway inhibition on piPSC pluripotency by activating phosphatidylinositol 3-kinase-protein kinase B(PI3K-AKT)signaling activity.Our research sheds new light on the mechanism by which OCT6 promotes PSC maintenance.

Fuzi promoted the formation of myelin sheath in Schwann cells induced by high glucose

OBJECTIVE Diabetic peripheral neuropathy(DPN)is the cause of considerable morbidity and mortality indiabetic patients.The loss of nerve fibersis the main pathological characteristics of the DPN and the pathway of Oct6-Krox20 plays an important role in the formation of myelin sheath.In our previous study,we found that Fuzi(Radix aconite lateral ispreparata)could significantly improve the nerve conduction deficits and thermal hypoalgesia deficits in the diabetic rats,but the underlying molecular mechanisms have not been established.The aim of this study is to investigate the expression of Oct6,Krox20,myelin basic protein(Mbp)and myelin protein zero(Mpz)in Schwann cells and analyze the effect of Fuzi in the formation of myelin sheath.METHODS There were six groups in the study.In the control group,thecells weresupplemented with normal cell culture medium.In the mannitol group,the cells were fed with normal glucose plus mannitol.In the model group,the cells were supplemented with high glucose medium(75 mmol·L-1).In the other group,the cells weretreated with high glucose medium(75mmol·L-1)plus different concentrations of Fuzi(0.1,1.0 and 10.0μg·mL-1).After three days,real-time PCR was used to detect gene expression.RESULTS Compared with the control group,the model group showed lowerexpression of Oct6,Krox20,Mbp and Mpz.In comparison to the model group,Fuzi(0.1,1.0and 10.0μg·mL-1)increased the expression of Oct6,Krox20,Mbp and Mpz.CONCLUSION These results demonstrate that Fuzi enhances the protein of myelin sheath through impacting the pathway of Oct6-Krox20.