马尾松松针提取物对人毛乳头细胞的保护作用及机制研究

doi:10.3969/j.issn.1001-1803.2023.01.009

Abstract

Abstract:

The effect and mechanism of Pinus massoniana pine needle extract on the activity of protecting human dermal papilla cells （HDP） were explored in this paper. The different extracts from pine needle were prepared with different solvents, and the content of active components was determined and analyzed by the corresponding determination method. HDP cells were induced by dihydrotestosterone（DHT）to establish androgenic alopecia cell model. CCK8 method was used to analyze the effect of pine needle extracts on the relative viability of the cells, and qPCR was adopted to investigate the effect of pine needle extract on regulating Wnt/β-Catenin signaling pathway by detecting the relative expression of VEGF, TGF-β2, β-Catenin, and DVL1 genes. UPLC-QTOF/MS was used to identify the components of the pine needle extract. The results show that the 50% alcohol extract of pine needle can significantly improve the relative activity of HDP cells （P<0.05）, increase the expression of VEGF mRNA （P<0.05）, decrease TGF-β2 mRNA relative expression （P<0.01）, and promote the expression of DVL1 mRNA and β-Catenin mRNA expression in the Wnt/β-Catenin signaling pathway（P<0.05, P<0.01）. In addition, a total of 36 components are identified from the 50% alcohol extract of pine needle, including flavonoids, lignans, alkaloids, and terpenes. Pinus massoniana pine needle extract has a protective effect on HDP cells, and its mechanism may be related to Wnt/β-Catenin signaling pathway.

Key words: Pinus massoniana pine needle, dihydrotestosterone, human dermal papilla cells, androgenetic alopecia

CLC Number:

TQ658

Wang Jiancheng, Xing Yan, Xu Ting, Wang Yue, Li Yongqiang, Zhang Limei. Protective effect and mechanism of Pinus massoniana pine needle extract in human dermal papilla cells[J].China Surfactant Detergent & Cosmetics, 2023, 53(1): 62-70.

Figures/Tables 11

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Tab. 3

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Tab. 4

Identification results of compounds in pine needle extract"

序号	保留时间/ min	中文名称	英文名称	分子式	[M-H] ^-	Score
1	1.06	杜仲甙Ⅱ	Eucommioside-Ⅱ	C₁₅H₂₆O₉	349.158 6	97.04
2	1.128	（-）-表儿茶素	（-）-Epicatechin	C₁₅H₁₄O₆	289.079 1	99.55
3	1.465	3，4-二羟基烯丙基苯-4-O-β-D-吡喃葡萄糖苷	3, 4-Dihydroxy-allylbenzene-4-O-beta-D-glucopyranoside	C₁₅H₂₀O₇	311.121 7	97.25
4	2.004	丁香苷	Syringafghanoside	C₂₅H₃₂O₁₁	507.194 7	98.64
5	2.273	柚皮素-7-O-葡萄糖苷	Naringenin-7-O-glucoside	C₂₁H₂₂O₁₀	433.122 7	98.23
6	2.812	1-（2’-γ-吡喃酮）-6-咖啡酰-α-D-吡喃葡萄糖	1-（2’-gamma-Pyranone）-6-caffeoyl-alpha-D-pyranoglucose	C₂₀H₂₀O₁₁	435.100 5	98.83
7	3.621	女贞果苷A	Lucidumoside A	C₂₅H₃₄O₁₂	525.205 5	99
8	3.823	花旗松素-3’-O-葡萄糖苷	Taxifolin-3’-O-glucoside	C₂₁H₂₂O₁₂	465.103 8	98.23
9	5.171	（-）-5’-甲氧基异戊烯雌酚-2a-O-β-D-木吡喃糖苷	（-）-5’-Methoxyisolariciresinol-2a-O-beta-D-xylopyranoside	C₂₆H₃₄O₁₁	521.210 9	99.13
10	5.845	6-O-E-阿魏基胡桃醇	6-O-E-Feruloylajugol	C₂₅H₃₂O₁₂	523.189 9	98.47
11	9.82	（-）-异紫杉醇-2a-O-β-D-木吡喃糖苷	（-）-Isolariciresinol-2a-O-beta-D-xylopyranoside	C₂₅H₃₂O₁₀	491.200 4	97.54
12	10.292	左旋马尾松树脂醇	Massoniresil	C₂₀H₂₄O₈	391.144 9	98.23
13	11.101	淫羊藿苷H1	Icariside H1	C₁₈H₂₆O₉	385.157 8	99.73
14	11.438	3，4’，5，7-四羟基黄酮-3-L-鼠李糖苷	3, 4’, 5, 7-Tetrahydroxyflavone-3-L-rhamnoside	C₂₁H₂₀O₁₀	431.106 1	95.52
15	11.707	（+）-1-羟基松脂醇-4’’-O-β-D-吡喃葡萄糖苷	（+）-1-Hydroxypinoresinol-4’’-O-beta-D-glucopyranoside	C₂₆H₃₂O₁₂	535.191	97.77
16	11.91	淫羊藿苷E6	Icariside E6	C₂₆H₃₆O₁₁	523.225 7	99.77
17	11.977	马尾松糖苷C	Massonianoside C	C₂₆H₃₄O₁₀	505.216	97.07
18	12.449	细叶皂苷D	Tenuifoliside D	C₁₈H₂₄O₉	383.142	99.54
19	15.212	（+）-环氧树脂醇	（+）-Epipinoresinol	C₂₀H₂₂O₆	357.142	98.99
20	16.761	槲皮素	Quercetin	C₁₅H₁₀O₇	301.042 5	98.54
21	21.276	鸢尾甲黄素B	Iristectorigenin B	C₂₉H₃₄O₁₇	653.180 2	98.97
22	25.454	6-乙酰基呋喃醇	6-Acetylfuranofukinol	C₁₇H₂₄O₄	291.168	98.55
23	27.341	5-羟基-6，7，3-三甲氧基黄酮-8- O-β-D-葡萄糖苷	5-Hydroxy-6, 7, 3-trimethoxyflavone-8-O-beta-D-glucoside	C₂₄H₂₆O₁₂	505.142 7	99.56
24	28.352	14-脱氧-11，12-二氢穿心莲内酯	14-Deoxy-11, 12-didehydroandrographolide	C₂₀H₂₈O₄	331.199 3	97.94
25	31.789	芹菜素-7-O-β-D-吡喃葡萄糖苷	Apigenin-7-O-β-D-glucopyranoside	C₃₉H₃₂O₁₄	723.179 5	99.69
26	33.541	10-姜二酮	10-Gingerdione	C₂₁H₃₂O₄	347.23	98.54
27	35.293	6-甲基二乙酸姜酯	6-Methylgingediacetate	C₂₂H₃₄O₆	393.235 8	98.91
28	36.978	12-甲氧基-8，11，13-枞三烯-7β，11-二醇	12-Methoxy-8, 11, 13-abietatriene-7beta, 11-diol	C₂₁H₃₂O₃	331.234 9	98.24
29	40.212	（-）-17-羟基-16β-考兰-19-油酸	（-）-17-Hydroxy-16beta-kauran-19-oicacid	C₂₀H₃₂O₃	319.235 1	99.53
30	42.908	高三尖杉酯碱	Homoharringtonine	C₂₉H₃₉NO₉	544.261 8	96.98
31	43.986	7β，12-二甲氧基-8，11，13-枞三烯-11-醇	7beta, 12-Dimethoxy-8, 11, 13-abietatrien-11-ol	C₂₂H₃₄O₃	345.250 5	99.27
32	47.962	6-甲基隐乙酰内酯	6-Methylcryptoacetalide	C₁₉H₂₄O₃	299.173 2	97.17
33	49.242	α，β-二半乳糖基-α’-亚麻酸甘油酯	alpha, beta-Digalactosyl-alpha’-linolenic-glyceride	C₃₃H₅₆O₁₄	675.367 8	98.55
34	49.646	9-D-羟基顺式-12-十八烯酸	9-D-Hydroxy-cis-12-octadecenoicacid	C₁₈H₃₄O₃	297.251	99.61
35	52.072	1-（3，4-亚甲基二氧苯基）-1E-十四烯	1-（3, 4-Methylenedioxyphenyl）-1E-tetradecene	C₂₁H₃₂O₂	315.240 7	99.18
36	53.892	补骨脂酚	Backuchiol	C₁₈H₂₄O	255.183 1	99.45

Tab. 4

References 21

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基因	上游引物	下游引物
VEGF	ATCGAGTACATCTTCAAGCCAT	GTGAGGTTTGATCCGCATAATC
TGFβ2	GCAAAGTTGTGAAAACAAGAGC	ATCCCAGGTTCCTGTCTTTATG
DVL1	TTTGAGAACATGAGCAATGACG	GGGACGGTGAAGTAGCTTC
β-Catenin	TGGATTGATTCGAAATCTTGCC	GAACAAGCAACTGAACTAGTCG
GAPDH	GGTGA AGGTC GGAGT CAACG	CAAAG TTGTC ATGGA TGHACC

提取物	总黄酮含量/ （mg/g，以芦丁计）	粗多糖含量/ （mg/g，以葡萄糖计）	原花青素含量/ （mg/g，以儿茶素计）	木脂素含量/ （mg/g，以新木脂素A计）
水提取物	55.9	148.3	69.5	183.0
30%乙醇提取物	76.2	117.2	97.8	200.4
50%乙醇提取物	91.2	63.8	102.4	228.3

Protective effect and mechanism of Pinus massoniana pine needle extract in human dermal papilla cells

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时间/min	A相比例/%	B相比例/%
0	80	20
25	60	40
30	45	55
60	30	70