珠黄复方制剂促进软组织和骨组织创口愈合的细胞学研究

李立峰; 杨华俊; 章巧丽; 来月丽; 赵彤

doi:10.16766/j.cnki.issn.1674-4152.001822

珠黄复方制剂促进软组织和骨组织创口愈合的细胞学研究

doi: 10.16766/j.cnki.issn.1674-4152.001822

杭州市富阳区第一人民医院口腔科，浙江杭州 311400

基金项目:

浙江省中医药科学研究基金项目 2018ZA105

详细信息

通讯作者:
李立峰，E-mail：liboboli1@126.com

中图分类号: R28 R783
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- 被引次数: 0
出版历程
- 收稿日期: 2020-09-27
- 网络出版日期: 2022-02-19

Zhuhuang compound preparation promotes the healing of soft and bone wounds in cellular level

Department of Stomatology, Fuyang People's Hospital, Hangzhou, Zhejiang 311400, China

摘要

摘要: 目的论证传统经方珠黄散的复方制剂在促进软组织和骨组织创口愈合方面的有效性，同时探讨复方制剂中5类中药的数量效应关系，为中药古方新的临床应用提供科学依据和推进方剂筛选与优化。方法以珠黄散为基本方，添加三七、接骨木、续断药材。采用均匀设计法配比药材用量，得到5种配方。将5组配方的药材制备水提物和醇提物干粉。用脂多糖(LPS)刺激RAW 309 Cr.1细胞后给予提取物检测炎症因子释放一氧化氮(NO)水平，采用噻唑蓝(MTT)法检测人脐静脉内皮细胞(HUVEC-12)增殖和成骨细胞(UMR-106)增殖水平。结果 5组复方中2、3、5号取物在1~5 μg/mL浓度下显示出44.3%~56.7%的NO的抑制作用，1、4号提取物在1~5 μg/mL浓度下对NO的抑制达46.7%~73.4%。4号提取物在3~4 μg/mL显示对HUVEC-12细胞有110%~128%的增殖作用，1号提取物在4 μg/mL显示110%~120%的增殖作用。5组复方提取物对UMR-106细胞增殖作用高于对照组的18%，浓度为4 μg/mL时增殖效果明显。结论 5组配方制剂体外细胞实验表现出单一或多重促进细胞增殖的作用。其中4号配方(珍珠8 g、牛磺8 g、三七6 g、断续15 g、接骨木9 g)显示出抑制炎症因子释放，对人脐静脉内皮细胞和成骨细胞增殖活性较强(P < 0.05)。传统经方珠黄散的复方制剂可能通过促进血管细胞和成骨细胞的增殖达到促进软组织和骨组织创口的愈合，4号配方抑制炎症因子释放，对人脐静脉内皮细胞增殖和成骨细胞增殖的影响显著。
- 珠黄复方制剂 /
- 抗炎 /
- 细胞增殖
Abstract: Objective To demonstrate the effectiveness of the traditional compound preparation of Zhuhuang powder in promoting wound healing of soft and bone as well as to explore the quantitative effect relationship of 5 kinds of traditional Chinese medicines in compound in the prescription, and provide scientific basis for the new clinical application of ancient Chinese medicines and screen new prescriptions. Methods Zhuhuang powder was used as the basic prescription, and Panax notoginseng, elderberry and intermittent medicinal materials were added. Five formulations were obtained by using uniform design method to proportionate the dosage of medicinal materials. The water extract and ethanol extract dry powder were prepared from five groups of prescriptions. RAW 309 Cr.1 cells stimulated with LPS were treated with these water extracts or ethanol extracts to detect inflammatory factors (NO) levels. The proliferation of human umbilical vein endothelial cells (HUVEC-12) and osteoblasts (UMR-106) were detected by methyl thiazolyl tetrazolium (MTT). Results The extract 2, 3 and 5 of 5 groups showed 44.3%-56.7% inhibition of NO expression at 1-5 μg/mL. The extract 1 and 4 showed 46.7%-73.4% inhibition of NO expression at 1-5 μg/mL. The extract 4 showed 110%-128% induction of the proliferation of HUVEC-12 cells at 3-4 μg/mL. The extract 1 showed 110%-120% proliferation at 4 μg/mL. Compared with control, all 5 groups showed 18% induction on the proliferation of UMR-106 cells, the proliferation effect was obvious at 4 μg/mL. Conclusion The five formulations showed single or multiple effects on cell proliferation in vitro. Formula No.4 (pearl 8 g, taurine 8 g, notoginseng 6 g, intermittent 15 g, elderberry 9 g) shows inhibition of inflammatory factors release and strong proliferation activity of human umbilical vein endothelial cells and osteoblasts (P < 0.05). The traditional prescription of Zhuhuang powder can promote the healing of soft tissue and bone tissue wounds. Formula 4 inhibits the release of inflammatory factors and has a significant effect on the proliferation of human umbilical vein endothelial cells and osteoblasts.
- Zhuhuang compound preparation /
- Anti-inflammation /
- Cell proliferation

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图 1 复方提取物对巨噬细胞RAW309 Cr.1释放NO的影响

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图 2 复方提取物对人脐静脉内皮细胞HUVEC-12增殖的影响

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图 3 复方提取物对成骨细胞UMR-106增殖的影响

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表 1 复方提取物均匀设计配方表 (g)

编号	清热抗炎药		三七活血化瘀药	接骨续筋药
编号	珍珠	牛黄	三七活血化瘀药	接骨木	续断
1	2.0	2.0	4.5	26.25	13.5
2	4.0	4.0	7.5	22.50	12.0
3	6.0	6.0	3.0	18.75	10.5
4	8.0	8.0	6.0	15.00	9.0
5	10.0	10.0	9.0	30.00	15.0

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表 2 复方提取物对RAW 309 Cr.1细胞释放NO的影响 (吸光度OD，x ±s)

配方		浓度(μg/mL)
配方		1	2	3	4	5
醇提取物	A1	1.511±0.121	1.709±0.113	1.804±0.106	1.812±0.127	1.821±0.103
	A2	1.201±0.133	1.227±0.132	1.303±0.103	1.316±0.124	1.331±0.107
	A3	1.207±0.134	1.302±0.103	1.304±0.122	1.317±0.117	1.333±0.128
	A4	1.722±0.141	1.724±0.142	1.803±0.150	1.814±0.143	1.873±0.102
	A5	1.211±0.127	1.228±0.104	1.303±0.112	1.315±0.130	1.337±0.131
水提取物	H6	1.211±0.120	1.308±0.131	1.321±0.142	1.357±0.139	1.359±0.142
	H7	1.799±0.103	1.821±0.111	1.853±0.105	1.884±0.118	1.889±0.128
	H8	1.214±0.155	1.219±0.136	1.313±0.141	1.351±0.151	1.359±0.103
	H9	1.811±0.121	1.831±0.135	1.862±0.126	1.884±0.117	1.902±0.129
	H10	1.215±0.111	1.220±0.106	1.313±0.115	1.353±0.123	1.357±0.101

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表 3 复方提取物对人脐静脉内皮细胞HUVEC-12增殖的影响 (吸光度OD，x ±s)

配方		浓度(μg/mL)
配方		1	2	3	4
醇提取物	A1	1.901±0.102	1.851±0.121	1.899±0.103	1.905±0.116
	A2	1.903±0.106	1.903±0.105	1.901±0.112	1.909±0.124
	A3	1.912±0.103	1.914±0.111	1.911±0.107	1.912±0.121
	A4	1.910±0.101	1.910±0.101	1.913±0.101	1.929±0.101
	A5	1.911±0.116	1.911±0.106	1.912±0.113	1.914±0.108
水提取物	H6	1.898±0.101	1.899±0.116	1.890±0.105	1.893±0.113
	H7	1.899±0.113	1.901±0.102	1.892±0.102	1.899±0.126
	H8	1.879±0.109	1.899±0.111	1.899±0.111	1.801±0.105
	H9	1.902±0.122	1.902±0.122	1.924±0.107	1.989±0.131
	H10	1.898±0.101	1.902±0.124	1.899±0.109	1.901±0.113

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表 4 复方提取物对成骨细胞UMR-106增殖的影响 (吸光度OD，x ±s)

配方		浓度(μg/mL)
配方		1	2	3	4
醇提取物	A1	1.871±0.107	1.876±0.103	1.899±0.125	1.897±0.115
	A2	1.888±0.104	1.889±0.101	1.901±0.112	1.901±0.120
	A3	1.879±0.106	1.888±0.121	1.911±0.107	1.912±0.107
	A4	1.911±0.130	1.922±0.104	1.930±0.112	1.934±0.115
	A5	1.889±0.104	1.890±0.105	1.912±0.113	1.913±0.111
水提取物	H6	1.859±0.121	1.861±0.105	1.850±0.117	1.855±0.105
	H7	1.859±0.105	1.861±0.101	1.882±0.118	1.889±0.116
	H8	1.861±0.113	1.866±0.115	1.859±0.109	1.868±0.113
	H9	1.912±0.121	1.933±0.107	1.954±0.115	1.979±0.104
	H10	1.898±0.111	1.801±0.131	1.879±0.119	1.899±0.122

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