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[1]胡松峰,郭蔓岑,金紅婷,等.基于網(wǎng)絡(luò)藥理學(xué)方法和動(dòng)物實(shí)驗(yàn)探討五福健膝方治療膝骨關(guān)節(jié)炎的作用機(jī)制[J].中醫(yī)正骨,2024,36(10):18-24.
　HU Songfeng,GUO Mancen,JIN Hongting,et al.Exploring the mechanism of Wufu Jianxi Fang(五福健膝方)against knee osteoarthritis based on the network pharmacology approach and animal experimentation[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2024,36(10):18-24.
點(diǎn)擊復(fù)制

基于網(wǎng)絡(luò)藥理學(xué)方法和動(dòng)物實(shí)驗(yàn)探討五福健膝方治療膝骨關(guān)節(jié)炎的作用機(jī)制()
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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第36卷
期數(shù):
2024年10期
頁(yè)碼:
18-24
欄目:
基礎(chǔ)研究
出版日期:
2024-10-20

文章信息/Info

Title:
Exploring the mechanism of Wufu Jianxi Fang(五福健膝方)against knee osteoarthritis based on the network pharmacology approach and animal experimentation
作者:
胡松峰1; 郭蔓岑2; 金紅婷3; 袁文華3
1.紹興市中醫(yī)院,浙江 紹興 312000; 2.浙江中醫(yī)藥大學(xué),浙江 杭州 310053; 3.浙江省中醫(yī)院,浙江 杭州 310006
Author(s):
HU Songfeng1; GUO Mancen2; JIN Hongting3; YUAN Wenhua3
1.Shaoxing Hospital of Traditional Chinese Medicine,Shaoxing 312000,Zhejiang,China 2.Zhejiang Chinese Medical University,Hangzhou 310053,Zhejiang,China 3.Zhejiang Provincial Hospital of Traditional Chinese Medicine,Hangzhou 310006,Zhejiang,China
關(guān)鍵詞:
骨關(guān)節(jié)炎; 膝;  五福健膝方;  腫瘤抑制蛋白p53;  網(wǎng)絡(luò)藥理學(xué);  動(dòng)物實(shí)驗(yàn)
Keywords:
osteoarthritis; knee;  Wufu Jianxi Fang;  tumor suppressor protein p53;  network pharmacology;  animal experimentation
摘要:
目的:探討五福健膝方治療膝骨關(guān)節(jié)炎(knee osteoarthritis,KOA)的作用機(jī)制。方法:①網(wǎng)絡(luò)藥理學(xué)研究。采用網(wǎng)絡(luò)藥理學(xué)方法篩選五福健膝方治療KOA的靶點(diǎn)。②動(dòng)物實(shí)驗(yàn)。將18只10周齡SPF級(jí)雄性C57BL/6小鼠隨機(jī)分為3組,每組6只。模型組和五福健膝方組小鼠采用內(nèi)側(cè)半月板失穩(wěn)術(shù)在右后肢構(gòu)建KOA模型,假手術(shù)組小鼠僅切開(kāi)對(duì)應(yīng)區(qū)域皮膚和關(guān)節(jié)囊后縫合。造模手術(shù)后第2天,五福健膝方組小鼠以五福健膝方湯劑(生藥濃度1.1 g·mL-1)灌胃,每次0.6 mL,每天1次,連續(xù)灌胃8周; 模型組和假手術(shù)組小鼠以等量生理鹽水灌胃。藥物干預(yù)結(jié)束后,收集小鼠右后肢膝關(guān)節(jié),分別進(jìn)行股骨遠(yuǎn)端Micro-CT掃描、組織病理學(xué)觀察(阿爾新藍(lán)-蘇木素/橙黃G染色)及免疫組織化學(xué)染色。免疫組織化學(xué)染色主要針對(duì)Ⅱ型膠原蛋白與網(wǎng)絡(luò)藥理學(xué)研究確定的核心靶點(diǎn)蛋白,測(cè)量Ⅱ型膠原蛋白表達(dá)陽(yáng)性區(qū)域的厚度(視為軟骨厚度),同時(shí)計(jì)算核心靶點(diǎn)蛋白陽(yáng)性細(xì)胞百分比。結(jié)果:①網(wǎng)絡(luò)藥理學(xué)研究結(jié)果。通過(guò)網(wǎng)絡(luò)藥理學(xué)研究確定的五福健膝方治療KOA的關(guān)鍵靶點(diǎn)基因15個(gè),其中TP53為核心靶點(diǎn)基因,其對(duì)應(yīng)的蛋白為P53蛋白。②動(dòng)物實(shí)驗(yàn)結(jié)果。股骨遠(yuǎn)端Micro-CT檢查結(jié)果顯示,模型組較假手術(shù)組骨表面積骨體積比值降低(P=0.028)、骨體積分?jǐn)?shù)升高(P=0.003),五福健膝方組較模型組骨表面積骨體積比值升高(P=0.004)、骨體積分?jǐn)?shù)降低(P=0.048)。膝關(guān)節(jié)軟骨組織阿爾新藍(lán)-蘇木素/橙黃G染色顯示,模型組小鼠軟骨缺失明顯,五福健膝方組小鼠軟骨丟失較模型組明顯改善。免疫組織化學(xué)染色結(jié)果顯示,模型組小鼠膝關(guān)節(jié)軟骨厚度較假手術(shù)組減小(P=0.001),五福健膝方組小鼠膝關(guān)節(jié)軟骨厚度較模型組增加(P=0.048); 模型組小鼠膝關(guān)節(jié)軟骨組織中P53陽(yáng)性細(xì)胞百分比較假手術(shù)組增加(P=0.000),五福健膝方組小鼠膝關(guān)節(jié)軟骨組織中P53陽(yáng)性細(xì)胞百分比較模型組減少(P=0.000)。結(jié)論:五福健膝方能夠有效延緩KOA模型小鼠的軟骨退變,其機(jī)制可能與其抑制P53蛋白表達(dá)有關(guān)。
Abstract:
Objective:To explore the mechanism of Wufu Jianxi Fang(五福健膝方,WFJXF)against knee osteoarthritis(KOA).Methods:①Network pharmacology research.The action targets of WFJXF against KOA were screened by using the network pharmacology approach.②Animal experimentation.Eighteen 10-week-old specific pathogen-free(SPF)-grade male C57BL/6 mice were selected and randomized into model group,WFJXF group and sham-operated group,6 cases in each group.The mice in model group and WFJXF group were subjected to destabilization of the medial meniscus(DMM)on the right hindlimbs for inducing KOA,while the ones in sham-operated group were merely incised the skin and joint capsule at the corresponding site and then sutured.On day 2 after the modeling surgery,the mice in WFJXF group were intervened by intragastric administration with WFJXF decoction(the crude drug concentration was 1.1 g/mL),once a day,0.6 mL at a time for consecutive 8 weeks; while the ones in model group and sham-operated group with the same dosage of normal saline.After the end of drug intervention,the mice were executed,and the knee joints were harvested from their right hindlimbs for scanning the distal femur by using Micro-CT,and observing the histopathological changes of knee articular cartilage tissues via alcian blue-hematoxylin/orange G(ABH/OG)staining,furthermore,the immunohistochemistry(IHC)staining was performed for detecting the typeⅡcollagen and core target proteins determined by network pharmacology analysis,and then the thickness of typeⅡcollagen-positive areas(regarded as the cartilage thickness)was measured,and the proportion of core target protein-positive cells was calculated.Results:①The results of network pharmacology research.As revealed by the network pharmacology analysis,15 key target genes of WFJXF against KOA were obtained,among which the TP53 was identified as the core target gene,with its corresponding protein being recognized as the P53.②The results of animal experimentation.As revealed by Micro-CT examination on distal femur,the ratio of bone surface(BS)to bone volume(BV)decreased,while the bone volume fraction(BVF)increased in model group compared to sham-operated group(P=0.028; P=0.003); whereas,the ratio of BS to BV increased and the BVF decreased in WFJXF group compared to model group(P=0.004; P=0.048).The ABH/OG staining results showed that the cartilages obviously lost in mice of model group,however,the cartilage loss was significantly improved in mice of WFJXF group compared with that of model group.Moreover,the results of IHC staining revealed that the thickness of knee cartilage decreased in mice of model group compared with that of sham-operated group(P=0.001),and it increased in mice of WFJXF group compared with that of model group(P=0.048); besides,the proportion of P53 positive cells in the knee articular cartilage tissues increased in mice of model group compared with that of sham-operated group(P=0.000),and it decreased in mice of WFJXF group compared with that of model group(P=0.000).Conclusion:WFJXF can effectively delay the cartilage degeneration in KOA model mice,which may work by inhibiting the expression of P53 protein.

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備注/Memo

備注/Memo:
基金項(xiàng)目:浙江省中醫(yī)藥科技計(jì)劃項(xiàng)目(2020ZA117); 浙江省基礎(chǔ)公益研究計(jì)劃項(xiàng)目(LTGY23H270008); 浙江省創(chuàng)傷性骨病診治中醫(yī)藥傳承創(chuàng)新團(tuán)隊(duì)項(xiàng)目(浙衛(wèi)發(fā)〔2023〕31號(hào))
通訊作者:袁文華 E-mail:[email protected]
更新日期/Last Update: 1900-01-01