壮药菲牛蛭提取物的美白作用及其机制初探

doi:10.3969/j.issn.1001-1803.2022.01.006

日用化学工业 ›› 2022, Vol. 52 ›› Issue (1): 35-43.doi: 10.3969/j.issn.1001-1803.2022.01.006

壮药菲牛蛭提取物的美白作用及其机制初探

何瑞源¹,王硕²,韦桂丽²,龚小妹²,吴佩莹³,缪剑华^1,^2,^4,^*()

1.广西中医药大学药学院,广西南宁 530200
2.广西药用植物园广西药用资源保护与遗传改良重点实验室,广西南宁 530023
3.广西医科大学,广西南宁 530021
4.广西药用植物园广西壮族自治区中药资源智慧创制工程研究中心,广西南宁 530023

收稿日期:2021-04-13 修回日期:2022-01-04 出版日期:2022-01-22 发布日期:2022-01-19
通讯作者: 缪剑华
基金资助:
广西创新驱动发展专项资金项目“药用植物大数据中心建设”资助(桂科AA18242040);广西自然科学基金(2020GXNSFAA259008);国家现代农业产业技术体系建设专项(CARS-21);对发展中国家科技援助项目(KY201904001)

Study of whitening effect and the mechanism of Zhuang medicine Poecilobdella manillensis extract

He Ruiyuan¹,Wang Shuo²,Wei Guili²,Gong Xiaomei²,Wu Peiying³,Miao Jianhua^1,^2,^4,^*()

1. School of Pharmacy, Guangxi University of Chinese Medicine, Nanning, Guangxi 530200, China
2. Guangxi Key Laboratory of Medicinal Resources Protection and Genetic Improvement, Guangxi Botanical Garden of Medicinal Plants, Nanning, Guangxi 530023, China
3. Guangxi Medical University, Nanning, Guangxi 530021, China
4. Guangxi Engineering Research Center of TCM Resource Intelligent Creation, Guangxi Botanical Garden of Medicinal Plants, Nanning, Guangxi 530023, China

Received:2021-04-13 Revised:2022-01-04 Online:2022-01-22 Published:2022-01-19
Contact: Jianhua Miao

摘要/Abstract

摘要：

研究壮药菲牛蛭提取物的美白作用及其机制。采用MTT法检测菲牛蛭提取物对黑色素瘤细胞（B16）的增殖抑制作用;采用分光光度法检测菲牛蛭提取物对B16细胞酪氨酸酶（TYR）活力和黑色素合成的影响以及对DPPH、·OH和·O₂^-三种自由基的清除能力和总抗氧化能力;采用紫外线照射法建立裸鼠皮肤色素沉着模型,Masson-Fontana银染法显示皮肤黑色素分布,并测定血清中TYR、超氧化物歧化酶（SOD）、丙二醛（MDA）、α-促黑色素生长激素（α-MSH）、碱性成纤维细胞生长因子（bFGF）、神经生长因子（NGF）、白介素（IL-1α）、肿瘤坏死因子（TNF-α）和γ-干扰素（IFN-γ）的水平。实验结果表明,菲牛蛭提取物能明显抑制B16细胞增殖和TYR活力,降低胞内黑色素含量;具有较强的清除自由基能力;能显著减少裸鼠皮肤中黑色素细胞的数量,提高裸鼠血清中SOD含量,降低MDA、α-MSH、bFGF、NGF、IL-1α、TNF-α和IFN-γ含量及TYR活力。壮药菲牛蛭提取物具有较好的美白作用,其作用机制可能与抑制TYR活性及抗氧化作用有关。

关键词: 菲牛蛭, 美白作用, 黑色素瘤细胞, 酪氨酸酶, 抗氧化

Abstract:

This research was aimed to study the whitening effect and mechanism of Zhuang medicine Poecilobdella manillensis extract. MTT method was used to detect the inhibitory effect of Poecilobdella manillensis extract on the proliferation of melanoma cells （B16）. Spectrophotometry was used to detect the effect of Poecilobdella manillensis extract on TYR activity and melanin synthesis in B16 cells, and the scavenging ability of DPPH, ·OH and ·O₂^- free radicals and the total antioxidant capacity were also detected. Based on an ultraviolet-irradiated nude mice skin pigmentation model, Masson Fontana silver staining was used to show the distribution of melanin in skin, and the levels of TYR, SOD, MDA, α-MSH, bFGF, NGF, IL-1α, TNF-α and IFN-γ in serum were also measured. The results show that the IC₅₀ of B16 cells proliferation are 230.66, 109.70, 53.18 μg/mL after Poecilobdella manillensis extract treatment at 24, 48 and 72 h, respectively. The IC₅₀ of arbutin at 24, 48 and 72 h are 1.11, 1.40 and 1.30 times higher than those of Poecilobdella manillensis extract, respectively. The non-cytotoxic concentration of Poecilobdella manillensis extract could significantly increase the tyrosinase activity and melanin content of B16 cells. The Poecilobdella manillensis extract has strong antioxidant ability in vitro. The IC₅₀ of DPPH, ·OH and ·O₂^- free radicals are 0.37, 0.38 and 0.43 g/L, respectively. The total antioxidant capacity of 0.8 g/L Poecilobdella manillensis extract is equivalent to 1.0 g/L vitamin E. Poecilobdella manillensis extract could significantly reduce the number of melanocytes in skin of nude mice, increase the content of SOD in serum, decrease the contents of MDA, α-MSH, bFGF, NGF, IL-1α, TNF-α and IFN-γ and TYR activity. Zhuang medicine Poecilobdella manillensis extract has a good whitening effect, and its mechanism may be related to the inhibition of tyrosinase activity and antioxidation capacity.

Key words: Poecilobdella manillensis, whitening effect, melanoma cells, tyrosinase, antioxidant

中图分类号:

TQ658

何瑞源,王硕,韦桂丽,龚小妹,吴佩莹,缪剑华. 壮药菲牛蛭提取物的美白作用及其机制初探[J]. 日用化学工业, 2022, 52(1): 35-43.

He Ruiyuan,Wang Shuo,Wei Guili,Gong Xiaomei,Wu Peiying,Miao Jianhua. Study of whitening effect and the mechanism of Zhuang medicine Poecilobdella manillensis extract[J]. China Surfactant Detergent & Cosmetics, 2022, 52(1): 35-43.

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	质量浓度/（μg/mL）	抑制率/%				质量浓度/（μg/mL）	抑制率/%
	质量浓度/（μg/mL）	24 h	48 h	72 h		质量浓度/（μg/mL）	24 h	48 h	72 h
菲牛蛭提取物	10	1.16±0.45	5.46±0.23	13.86±0.75	熊果苷	10	1.89±1.03	7.21±1.68	10.71±0.05
	20	3.01±0.59	13.49±3.37	21.41±2.15		20	3.96±1.84	10.53±0.47	20.56±0.32
	40	15.96±3.48	30.87±3.03	44.99±1.68		40	12.23±2.81	23.15±0.46	40.73±2.12
	60	16.75±2.56	34.17±4.46	48.32±2.14		60	14.76±2.52	29.28±2.22	46.81±3.87
	80	17.34±2.17	38.72±2.78	55.82±1.38		80	18.88±1.12	30.60±2.05	48.88±3.12
	100	26.35±4.69	47.02±5.67	76.37±7.35		100	25.06±4.28	42.37±5.17	61.25±4.06
	IC₅₀	230.66	109.70	53.18		IC₅₀	255.40	153.19	68.96

	质量浓度/（μg/mL）	48 h抑制率/%		质量浓度/（μg/mL）	48 h抑制率/%
熊果苷	14	10.96±1.25	菲牛蛭提取物	14	21.63±1.44^△△
	7	7.13±0.94		7	11.19±1.11^△△
	3.5	5.64±0.54		3.5	8.24±1.20^△

	质量浓度/（μg/mL）	48 h抑制率/%		质量浓度/（μg/mL）	48 h抑制率/%
熊果苷	14	7.12±1.03	菲牛蛭提取物	14	9.90±0.93^△
	7	5.81±1.09		7	8.03±0.21^△
	3.5	4.61±0.36		3.5	4.98±0.39

	质量浓度/（g/L）	清除率/%				质量浓度/（g/L）	清除率/%
	质量浓度/（g/L）	DPPH	·OH	·O₂^-		质量浓度/（g/L）	DPPH	·OH	·O₂^-
菲牛蛭提取物	0.1	21.23±1.73	20.23±2.55	23.43±3.21	V_E	0.1	15.36±1.54	15.37±1.95	20.35±2.31
	0.2	30.32±1.44	30.78±2.22	35.2±2.13		0.2	21.05±2.57	27.69±2.38	25.67±1.58
	0.3	38.16±2.31	38.16±1.89	40.53±1.56		0.3	28.16±2.17	31.44±1.96	31.28±3.25
	0.4	46.76±1.42	44.58±2.07	48.78±1.77		0.4	36.55±3.53	39.08±2.39	36.25±3.50
	0.5	56.32±2.09	50.32±1.07	56.32±2.33		0.5	43.67±2.13	44.27±1.85	41.98±2.05
	0.6	67.19±2.19	63.73±1.87	66.77±1.99		0.6	50.37±3.45	56.38±2.37	47.38±2.66
	0.7	73.66±1.89	70.67±1.53	73.87±2.07		0.7	55.69±2.67	62.34±2.22	52.87±1.89
	0.8	75.39±2.33	72.72±2.07	75.56±1.53		0.8	68.25±3.41	66.02±3.57	59.13±2.43
	0.9	78.13±1.17	73.32±1.02	76.43±1.63		0.9	74.01±2.57	70.08±2.59	68.24±2.33
	1.0	80.12±1.87	78.21±1.89	77.25±2.78		1.0	76.25±3.74	70.39±2.58	69.52±1.66

	质量浓度/（g/L）	总抗氧化能力/mL		质量浓度/（g/L）	总抗氧化能力/mL
菲牛蛭提取物	0.1	1.25±0.31	V_E	0.1	0.58±0.38
	0.2	2.66±0.61		0.2	0.95±0.15
	0.3	3.18±0.58		0.3	1.28±0.34
	0.4	3.96±0.62		0.4	1.89±0.21
	0.5	4.51±0.45		0.5	2.38±0.47
	0.6	5.27±0.42		0.6	3.66±0.48
	0.7	5.83±0.47		0.7	4.58±0.34
	0.8	6.33±0.58		0.8	5.13±0.61
	0.9	6.99±0.61		0.9	5.97±0.32
	1.0	7.15±0.56		1.0	6.52±0.57

壮药菲牛蛭提取物的美白作用及其机制初探

Study of whitening effect and the mechanism of Zhuang medicine Poecilobdella manillensis extract

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组别	给药质量浓度/（g/L）	MDA/（nmol/mL）	SOD/（nmol/mL）	TYR/（nmol/mL）
空白组	—	9.54±0.98	127.76±9.48	77.86±2.25
模型组	—	14.95±1.59^△	106.78±6.05^△	90.36±1.69^△
阳性组	0.100	10.38±1.39^**	120.92±10.10^**	80.12±2.31^**
低剂量组	0.025	14.70±0.59	105.35±8.98	87.22±2.09
中剂量组	0.050	12.79±1.57	111.55±5.87	81.09±3.31^**
高剂量组	0.100	9.89±1.13^**	122.98±8.26^**	79.87±1.49^**

组别	给药质量浓度/（g/L）	α-MSH/（nmol/mL）	bFGF/（nmol/mL）	NGF/（nmol/mL）
空白组	—	25.63±3.69	58.39±8.21	103.62±9.12
模型组	—	39.65±6.39^△△	89.36±7.01^△△	134.25±6.20^△△
阳性组	0.100	16.99±3.25^**	61.37±5.23^**	90.32±8.65^**
低剂量组	0.025	36.31±6.92	88.24±7.25	128.36±6.38
中剂量组	0.050	28.36±5.03^*	82.33±7.20	113.25±5.22^*
高剂量组	0.100	20.50±3.27^**	68.61±5.07^**	100.58±6.27^**

组别	给药质量浓度/（g/L）	IL-1α/（nmol/mL）	TNF-α/（nmol/mL）	IFN-γ/（nmol/mL）
空白组	—	38.88±5.37	22.22±5.48	160.62±10.85
模型组	—	79.65±5.21^△△	47.66±6.31^△△	199.45±10.65^△△
阳性组	0.100	41.31±3.63^**	27.11±3.15^**	137.61±12.15^**
低剂量组	0.025	74.32±6.17	43.39±5.92	189.24±11.93
中剂量组	0.050	70.08±8.67^*	38.71±6.71^*	176.46±10.26^**
高剂量组	0.100	45.40±3.90^**	26.86±3.29^**	152.38±11.93^**

组别	给药质量浓度/（g/L）	阳性面积比例（×100）	MOD（×100）	IOD（×100）
空白组	—	0.05±0.03	10.32±3.42	0.13±0.10
模型组	—	0.80±0.34^△	14.03±5.61^△	0.25±0.04^△
阳性组	0.100	0.65±0.24^*	12.09±4.37^*	0.20±0.09
低剂量组	0.025	0.78±0.31	13.99±7.52	0.23±0.8
中剂量组	0.050	0.66±0.12	13.78±2.61	0.18±0.05
高剂量组	0.100	0.30±0.23^*	11.02±6.32^*	0.15±0.04^*

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