本文關(guān)鍵詞： 化痰除濕祛瘀劑 人骨關(guān)節(jié)炎 細(xì)胞凋亡　出處：《中華中醫(yī)藥學(xué)刊》2017年06期 　論文類(lèi)型：期刊論文

【摘要】：目的:研究化痰除濕祛瘀劑對(duì)人骨關(guān)節(jié)炎(osteoarthritis,OA)軟骨細(xì)胞凋亡的影響。方法:(1)在蘇州中醫(yī)院骨傷科協(xié)助下,取因外傷急性截肢患者的軟骨組織以及全膝關(guān)節(jié)置換手術(shù)OA患者的軟骨組織,采用酶消化培養(yǎng)法培養(yǎng)人軟骨細(xì)胞。采用甲苯胺藍(lán)染色鑒定細(xì)胞來(lái)源,觀察顯微鏡觀察其形態(tài)變化。(2)將軟骨細(xì)胞分成OA軟骨細(xì)胞組、OA軟骨細(xì)胞組+化痰除濕祛瘀劑(0.016、0.08、0.4、2、10 mg/mL)組,根據(jù)分組情況分別培養(yǎng)24 h或48 h后,應(yīng)用MTT法比較各組細(xì)胞的增殖情況。(3)根據(jù)增殖結(jié)果將軟骨細(xì)胞分成OA軟骨細(xì)胞組、OA軟骨細(xì)胞組+化痰除濕祛瘀劑(0.05、0.1、2 mg/mL)組,用原位末端標(biāo)記法(TUNEL)檢測(cè)各組細(xì)胞凋亡情況。結(jié)果:(1)成功建立了人骨關(guān)節(jié)炎軟骨細(xì)胞有限細(xì)胞系,細(xì)胞生長(zhǎng)狀態(tài)良好;甲苯胺藍(lán)證明所培養(yǎng)的細(xì)胞為來(lái)自中胚層的軟骨細(xì)胞。(2)與對(duì)照組比較(24 h+),不同濃度的化痰除濕祛瘀劑對(duì)OA軟骨細(xì)胞活力無(wú)影響。與對(duì)照組(OD值:0.393±0.010,λ=450 nm)比較(24 h+),化痰除濕祛瘀劑(1 mg/mL,OD值:0.441±0.009)和(2mg/mL,OD值:0.430±0.012)組可明顯促進(jìn)軟骨細(xì)胞增殖(均P0.001)。(3)OA軟骨細(xì)胞經(jīng)TUNEL標(biāo)記后的結(jié)果顯示,對(duì)照組的凋亡指數(shù)為(42.21±2.67)%,化痰除濕祛瘀劑(0.1 mg/mL,37.14%±4.01%;2 mg/mL,34.21%±5.00%)可明顯抑制軟骨細(xì)胞早期凋亡與對(duì)照組比較(P0.05或P0.01)。結(jié)論:化痰除濕祛瘀劑促進(jìn)細(xì)胞增殖,抑制軟骨細(xì)胞過(guò)度凋亡,這可能是抑制軟骨破壞的重要機(jī)制之一。
[Abstract]:Objective: to study the effect of phlegm removing dampness and removing blood stasis on the apoptosis of chondrocytes in osteoarthritis osteoarthritis OAA. Methods: 1) with the assistance of Department of Orthopedics and Trauma, Suzhou traditional Chinese Medicine Hospital. The cartilage tissue of acute amputation due to trauma and the cartilage tissue of OA patients undergoing total knee arthroplasty were collected and cultured by enzyme digestion. The origin of the cells was identified by toluidine blue staining. The chondrocytes were divided into OA chondrocyte group and OA chondrocyte group. The 10 mg / mL group was cultured for 24 h or 48 h according to the group condition. According to the results of proliferation, chondrocytes were divided into OA chondrocyte group and OA chondrocyte group. In 2 mg / mL group, Tunel was used to detect the apoptosis of human osteoarthritis chondrocytes. Results: 1) Human osteoarthritis chondrocyte limited cell line was successfully established. Cell growth was good; Toluidine blue showed that the cultured cells were chondrocytes from mesoderm. The activity of OA chondrocytes was not affected by different concentrations of phlegm removing dampness and removing blood stasis, and the OD value of OA chondrocytes was 0.393 鹵0.010, 位 450nm compared with the control group for 24 h). The OD value of phlegm removing dampness and removing blood stasis was 0. 441 鹵0. 009 and the OD value of 1 mg / mLX was 0. 441 鹵0. 009) and 2 mg / mL. OD: 0.430 鹵0.012) group could significantly promote the proliferation of chondrocytes (all P0.001 ~ (3) OA chondrocytes were labeled with TUNEL. The apoptotic index of the control group was 42.21 鹵2.67, and that of the phlegm removing dampness and removing blood stasis was 37.14% 鹵4.01%, and that of removing phlegm and removing dampness and removing blood stasis was 37.14% 鹵4.01; 2 mg/mL. (34.21% 鹵5.00) can significantly inhibit the early apoptosis of chondrocytes compared with the control group, P0.05 or P0.01.Conclusion: phlegm removing dampness and removing stasis can promote the proliferation of chondrocytes. Inhibition of excessive apoptosis of chondrocytes may be one of the important mechanisms to inhibit cartilage damage.
【作者單位】： 蘇州吳門(mén)醫(yī)派研究院;上海中醫(yī)藥大學(xué)附屬曙光醫(yī)院;蘇州市中醫(yī)醫(yī)院骨傷科;
【基金】：蘇州市科技局計(jì)劃項(xiàng)目(SYS201420)
【分類(lèi)號(hào)】：R274.9

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