南京中医药大学
纸质出版:2020
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[1]赵凡,尚志远,瞿融.黄芩含药血清对神经干细胞分化钠离子通道基因及cAMP/PKA信号通路的影响[J].中医杂志,2020,61(16):1444-1448.
ZHAO Fan, SHANG Zhiyuan, QU Rong. Effects of the Serum Containing Radix Scutellariae- on Sodium Channel Genes and cAMP/PKA Signaling Pathway of Neural Stem Cells Differentiation[J]. Journal of Traditional Chinese Medicine2020, 61(16): 1444-1448.
[1]赵凡,尚志远,瞿融.黄芩含药血清对神经干细胞分化钠离子通道基因及cAMP/PKA信号通路的影响[J].中医杂志,2020,61(16):1444-1448. DOI: 10.13288/j.11-2166/r.2020.16.016.
ZHAO Fan, SHANG Zhiyuan, QU Rong. Effects of the Serum Containing Radix Scutellariae- on Sodium Channel Genes and cAMP/PKA Signaling Pathway of Neural Stem Cells Differentiation[J]. Journal of Traditional Chinese Medicine2020, 61(16): 1444-1448. DOI: 10.13288/j.11-2166/r.2020.16.016.
目的研究黄芩含药血清对神经干细胞分化的影响及可能机制。方法分离培养原代乳大鼠神经干细胞
利用免疫荧光法进行鉴定。SD大鼠分别给予500和1000mg/kg黄芩水煎剂每日灌胃1次
连续7天后制备黄芩高、低剂量含药血清。黄芩低、高含药血清均设0.1%、1%、10%三个浓度干预神经干细胞
进行试验药物浓度筛选。收集神经干细胞以每孔1×10
5
个接种于24孔板中
实验分为对照组、黄芩含药血清组和含药血清加抑制剂组
每组3个复孔。对照组每孔加入10μl空白大鼠血清
黄芩含药血清组每孔加入10μl筛选浓度的黄芩含药血清
含药血清加抑制剂组每孔加入10μl筛选浓度的黄芩含药血清及1μl浓度为10mmol/L的H89溶液
隔天换液
共分化7天。比较各组细胞中双肾上腺皮质激素(DCX)、神经元核心抗原(NeuN)细胞增长率;检测细胞中钠离子通道亚型SCN1A、SCN2A、SCN3A、SCN8A mRNA表达
蛋白激酶A(PKA)、磷酸化蛋白激酶A(p-PKA)蛋白表达
细胞上清中腺苷酸环化酶(AC)、环磷酸腺苷(cAMP)浓度。结果筛选出1%黄芩高剂量含药血清用于后续实验。与对照组比较
黄芩含药血清组细胞中DCX、NeuN细胞增长率增加
SCN1A、SCN2A、SCN3A、SCN8A mRNA表达增加
p-PKA/PKA值及AC、cAMP浓度增加(P
<
0.05或P
<
0.01);与黄芩含药血清组比较
含药血清加抑制剂组上述各指标均不同程度降低(P
<
0.01)。结论黄芩含药血清可促进海马神经干细胞分化
其机制可能与促进钠离子通道基因、cAMP/PKA通路酶及蛋白的表达有关。
Objective To study the effects and possible mechanism of Huangqin(Radix Scutellaria)-containing serum on differentiation of neural stem cells. Methods Primary rat neural stem cells were isolated
cultured and identified by immunofluorescence. SD rats were given 500 and 1000 mg/kg of Huangqin Decoction(黄芩水煎剂) once a day for gavage. After 7 consecutive days
high-dose and low-dose Radix Scutellariae-containing serum were prepared. The low and high drug-containing serums were set at 3 concentrations of 0.1%
1%
and 10% to intervene neural stem cells
and the test drug concentration was screened. Neural stem cells were collected and inoculated into a 24-well plate with 1×10
5
per well. The experiment was divided into a control group
a drug-containing serum group
and a drug-containing serum plus inhibitor group
with 3 replicate wells per group. The control group was added with 10μl of blank rat serum per well
the drug-containing serum group was added with 10μl of screening concentration of Radix Scutellariae-containing serum per well
the drug-containing serum plus inhibitor group was added with 10μl of screening concentration of Radix Scutellariae-containing serum per well. In the drug-containing serum plus inhibitor group
10μl of Radix Scutellariae containing serum and 1μl of H89 solution at a concentration of 10 mmol/L were added to each well
and the fluid was changed every other day for 7 days. The cell growth rates of the double adrenocortical hormone(DCX) and neuronal core antigen(NeuN) in each group of cells were compared; the expre-ssion of sodium channel subtypes SCN1 A
SCN2 A
SCN3 A
SCN8 A mRNA
protein kinase A(PKA)
phosphate in the cells Protein kinase A(p-PKA) protein expression
concentration of adenylate cyclase(AC) and cAMP in cell supernatant were detected. Results 1% Radix Scutellariae high-dose serum was screened for subsequent experiments. Compared with the control group
the growth rate of DCX and NeuN cells in the drug-containing serum group cells increased
SCN1 A
SCN2 A
SCN3 A
SCN8 A mRNA expression increased
p-PKA/PKA value and AC
cAMP concentration increased(P
<
0.05 or P
<
0.01); compared with the drug-containing serum group
the above-mentioned indexes of the drug-containing serum plus inhibitor group were reduced to varying degrees(P
<
0.01). Conclusion The Radix Scutellariae-containing serum can promote the differentiation of hippocampal neural stem cells. The mechanism may be related to the promotion of the expression of sodium channel genes
cAMP/PKA pathway enzymes and proteins.
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