In vitro effects of sodium hyaluronate on the proliferation and the apoptosis in chondrocytes from patients with Kashin-Beck disease and osteoarthritis

Objective：To identify the in vitro effects of sodium hyaluronate（HA） on the proliferation and the apoptosis of chondrocytes from patients with Kashin-Beck disease（KBD） and osteoarthritis（OA）. Methods：Samples of articular cartilages from KBD and OA patients, as well as healthy volunteers（6 subjects in each of the 3 groups） were dissected, digested with collagenase and the cells cultured in monolayers. Chondrocytes from each sample were assigned to an untreated group and two HA-treated groups： H0（no HA）, H100（HA, 0.1 g/L） and H500（HA, 0.5 g/L）. The first passage chondrocytes were used to observe proliferation using the MTT assay, and apoptosis by flow cytometry through Annexin V/PI staining. Results：HA promoted proliferation of chondrocytes in all the three groups, and.in KBD and OA groups, for cells cultured for 4 and 6 days, H500 significantly promoted the cell proliferation. The apoptotic rates of both KBD and OA group chondrocytes were in the order H500 〈 HA100 〈 H0. Conclusion：Sodium hyaluronate administration has a dosedependent in vitro effect to promote proliferation and inhibit apoptosis of chondrocytes from patients with KBD and OA.

Effect of sodium hyaluronate combined with rehabilitation training on knee joint injury caused by golf

BACKGROUND In high-intensity sports like golf,knee joints are prone to injury,leading to pain,limited mobility,and decreased quality of life.Traditional treatment methods typically involve rehabilitation exercises,but their effectiveness may be limited.In recent years,sodium hyaluronate has emerged as a widely used biomedical material in the treatment of joint diseases.AIM To explore the effect of sodium hyaluronate combined with rehabilitation training on pain degree,flexion range of motion and motor function of knee joint injured by golf.METHODS Eighty patients with knee joint injury caused by golf were randomly divided into control(group B)and observation group(group A).The group B was treated with rehabilitation training,and the group A was treated with sodium hyaluronate combined with rehabilitation training.The clinical efficacy,range of motion and function of knee joint,quality of life and inflammatory factors were compared.RESULTS The excellent and good rate of rehabilitation in the group A was raised than group B.At 6 weeks and 3 months after treatment,the range of motion of the two groups was raised than that before treatment,and that of the group A was raised than group B.After treatment,the scores of Lysholm and International Knee Documentation Committee(IKDC)in the group A were raised,and those in the group A were raised than group B.The VAS score of the two groups was reduced than that of the group B,and the SF-36 score of the group A was reduced than group B.The interleukin(IL)-1β,IL-8 and tumor necrosis factor-αin the two groups were reduced,and those in the group A were reduced than group B.CONCLUSION Sodium hyaluronate combined with rehabilitation training has a good clinical effect in the treatment of patients with knee joint injury caused by golf,which relieve pain,maintain knee joint function and improve patients'life quality.

Efficacy of recombinant human epidermal growth factor plus sodium hyaluronate eye drops in diabetic dry eye post-cataract surgery

BACKGROUND Dry eye syndrome(DES)after diabetic cataract surgery can seriously affect the patient’s quality of life.Therefore,effective alleviation of symptoms in patients with this disease has important clinical significance.AIM To explore the clinical effect of recombinant human epidermal growth factor(rhEGF)plus sodium hyaluronate(SH)eye drops on DES after cataract surgery in patients with diabetes.METHODS We retrospectively evaluated 82 patients with diabetes who experienced DES after cataract surgery at Tianjin Beichen Hospital,Affiliated Hospital of Nankai University between April 2021 and April 2023.They were classified into an observation group(42 cases,rhEGF+SH eye drops)and a control group(40 cases,SH eye drops alone),depending on the different treatment schemes.The therapeutic efficacy,dry eye symptom score,tear film breakup time(TFBUT),basic tear secretion score[assessed using Schirmer I test(SIt)],corneal fluorescein staining(FL)score,tear inflammatory markers,adverse reactions during treat-ment,and treatment satisfaction were compared between the two groups.RESULTS Therapeutic efficacy was higher in the observation group compared with the control group.Both groups showed improved TFBUT and dry eye,as well as improved SIt and FL scores after treatment,with a more pronounced improvement in the observation group.Although no marked differences in adverse reactions were observed between the two groups,treatment satisfaction was higher in the observation group.CONCLUSION rhEGF+SH eye drops rendered clinical benefits to patients by effectively ameliorating dry eye and visual impairment with favorable efficacy,fewer adverse reactions,and high safety levels.Thus,this treatment should be promoted in clinical practice.

Efficacy and safety of 0.4 percent sodium hyaluronate for endoscopic submucosal dissection of gastric neoplasms

AIM:To evaluate the efficacy and safety of sodium hyaluronate solution(SH) in endoscopic submucosal dissection(ESD) of gastric neoplasms.METHODS:A prospective multicenter randomized,double blind,controlled trial was designed and utilized in this study.A total of 76 patients with 5-20 mm sized gastric neoplasms were enrolled at three academic hospitals in South Korea from June 2011 to October 2011.Patients were randomly assigned to the 0.4% sodium hyaluronate or control groups.All lesions underwent endoscopic ESD.ESD was performed with 0.4%SH and normal saline(NS) solution for submucosal injection.Efficacy was assessed using en bloc resection and the number of additional injections.Secondary evaluation variables were the volume of injection material,steepness of mucosal elevation,bleeding rate,procedural time and operator satisfaction.Finally,the safety was assessed by analyzing adverse events during the study.RESULTS:The usefulness rate in the 0.4%SH group and the controlled group had statistically significant difference under intention to treat(ITT) analysis(90.91% vs 61.11% P = 0.0041).Under per protocol(PP),the usefulness rate is statistically significant different(93.10% vs 61.76%,P = 0.0036).The difference in volume of the solution injected between 0.4%SH group and the controlled group and NS group was also statistically significant under intention to treat and per protocol analysis(ITT:0.03 ± 0.02 mL vs 0.06 ± 0.03 mL,P = 0.0003,PP:0.03 ± 0.02 mL vs 0.06 ± 0.03 mL,P = 0.0004).Satisfaction above the grade good was significantly higher in the SH group under intention to treat and per protocol analysis(ITT:90.91% vs 61.11%,P = 0.0041,PP = 93.11% vs 61.77%,P = 0.0022).Adverse events above grade 3 were not noticed in either group.All adverse events were treated and were judged as not associated with the submucosal injection solutions.CONCLUSION:0.4%SH solution is a safe and effective agent that doesn't cause any significant adverse events and is useful for submucosal injection during ESD.

Low concentration of sodium hyaluronate temporarily elevates the tear film lipid layer thickness in dry eye patients with lipid deficiency

AIM： To investigate the effects of different concentrations of artificial tears on lipid layer thickness （LLT） and blink rate （BR） in dry eye patients. METHODS： This study included 106 eyes of 58 patients with dry eye. The lipid deficiency type was defined as the LLT baseline 〈75 nm. The LLT and BR were measured at baseline and 1, 5 and 15min after the instillation of 0.1% or 0.3% sodium hyaluronate （SH） eye drops by using the LipiView ocular surface interferometer. RESULTS： In the lipid deficiency group, the LLT increased from baseline at 1rain post instillation. The LLT after the instillation of 0.1% SH was significantly higher than that after the instillation of 0.3% SH （P〈0.001）. The LLT returned to baseline at 15min post instillation of 0.1% SH and at 5min post instillation of 0.3% SH. In the non-lipid deficiency group, the LLT decreased from baseline at lmin and returned to baseline at 5rain for both treatments. The BRs were not significantly different at different time points for both treatments. CONCLUSION： SH eye drops induce a short-term increase in LLT of patients with lipid deficiency. A low concentration of artificial tears have a stronger effect than a high concentration of artificial tears on the increase in LLT. in comparison, SH eye drops induce a transient and slight decrease in LLT of patients without lipid deficiency. A low concentration of artificial tears might be better for patients with lipid deficiency.

Determination of sodium hyaluronate in pharmaceutical formulations by HPLC–UV

A liquid chromatography （HPLC） method with UV detection was developed for determination of sodium hyaluronate in pharmaceutical formulation. Sodium hyaluronate is a polymer of disaccharides, composed of D-glucuronic acid and D-N-acetylglucosamine, linked via alternating 13-1, 4 and D-l, 3 glycosidic bonds. Being a polymer compound it lacks a UV absorbing chromophore. In the absence of a UV absorbing chromophore and highly polar nature of compound, the analysis becomes a major challenge. To overcome these problems a novel method for the determination of sodium hyaluronate was developed and validated based on size exclusion liquid chromatography （SEC） with UV detection. An isocratic mobile phase consisting of buffer 0.05 M potassium dihydrogen phosphate, pH adjusted to 7.0 using potassium hydroxide （10%） was used. Chromatography was carried out at 25 ~C on a BioSep SEC $2000, 300 mm ~ 7.8 mm column. The detection was carded out using variable wavelength UV-vis detector set at 205 nm. The compounds were eluted isocratically at a steady flow rate of 1.0 mL/min. Sodium hyaluronate retention time was about 4.9 min with an asymmetry factor of 1.93. A calibration curve was obtained from 1 to 38 g/mL （r〉0.9998）. Within-day % RSD was 1.0 and between-day % RSD was 1.10. Specificity/selectivity experiments revealed the absence of interference from excipients, recovery from spiked samples for sodium hyaluronate was 99-102. The developed method was applied to the determination of sodium hyaluronate in pharmaceutical drug substance and product.

Determination of sodium hyaluronate with some basic bisphenylnaphthylmethane dyes by resonance Rayleigh scattering method

In an acetic acid–sodium acetate buffer solution of pH 3.6–6.8, a compound complex was formed between sodium hyaluronate (abbreviated as SH) and some basic bisphenylnaphthylmethane dyes, leading to a great enhancement of the intensity of resonance Rayleigh scattering (RRS) and giving a new RRS spectrum, with its maximum scattering peak near 280 nm. It was also found that the intensity of RRS was directly proportional to the concentration of SH near the range between 0 and 3.0 mg/L. Based on these facts, a sensitive method for the determination of SH has been established. The method had good selectivity, and has been used for the determination of total amounts of SH in samples with satisfactory results. For the NB–SH system, the detection limit of SH was down to 13.7 ng/mL.

Anti-scarring effect of sodium hyaluronate at filtration pathway after filtering surgery in rabbits

AIM: To investigate the anti-scarring effect of sodium hyaluronate(HA) at filtration pathway after filtering surgery in a rabbit model.METHODS: Fifteen healthy adult New Zealand white rabbits were selected for trabeculectomy in both eyes. The right eyes were used as HA group with 0.1 m L HA injected into the anterior chamber at the end of the operation;the left eyes were used with 0.1 m L sodium lactate Ringer’s solution(RS) injected into the anterior chamber as RS group. Intraocular pressure(IOP), filtering blebs morphology, inflammatory reaction and complications were observed at the 7, 60, and 90 d after surgery.RESULTS: One day after surgery, the IOP of HA and RS groups were 12.75±1.92 and 10.50±1.59 mm Hg(P=0.005). At the 7;day postoperative, the filtering blebs of each group were functional type and TGF-β expression was significantly difference in both groups(0.10±0.01 vs 0.14±0.02, P=0.024). After 60 d of the operation, all filtering blebs were scarring and alpha-smooth muscle actin(α-SMA) expression was significantly difference in both groups(0.40±0.04 vs 0.35±0.02, P=0.032). α-SMA positive cells were mainly distributed in the junction of conjunctiva and sclera and around the blood vessels. The collagen volume fraction(CVF) of HA and RS group was(75.49±7.01)% and(79.93±5.35)%(P=0.044). On the 90;day after the operation, CVF was(82.57±5.19)% and(88.08±1.75)% in HA and RS groups(P=0.036). There was no α-SMA positive cell in HA group, while a few positive cells were observed in RS group(P=0.000).CONCLUSION: HA has effect of anti-scar and antiinflammation on filtration pathway after filtering surgery within 3 mo by inhibiting fibroblast proliferation and collagen deposition.

Effect of arthroscopic debridement combined with intra-articular sodium hyaluronate injection on the Wnt/β-catenin pathway in joint fluid of patients with knee osteoarthritis

Objective: To investigate the effect of arthroscopic debridement combined with intra-articular sodium hyaluronate injection on the Wnt/β-catenin pathway in joint fluid of patients with knee osteoarthritis. Methods: Patients undergoing arthroscopic debridement for knee osteoarthritis in the hospital between June 2014 and April 2017 were selected as the research subjects and randomly divided into the observation group who accepted arthroscopic debridement combined with intra-articular sodium hyaluronate injection and the control group who accepted arthroscopic debridement alone. The Wnt/β-catenin pathway molecules as well as downstream collagen metabolism molecules and bone metabolism molecules in the joint fluid were measured before surgery and 7 d after surgery. Results: 7 d after surgery, Wnt1, Wnt5a, Wnt7b, β-catenin, MMP1, MMP13, CTX-I, CTX-II, ADAMTS4, ADAMTS5, RANK, RANKL and TRACP5b mRNA expression in joint fluid of both groups of patients were higher than those before surgery whereas OPG and OC contents were lower than those before surgery, and Wnt1, Wnt5a, Wnt7b, β-catenin, MMP1, MMP13, CTX-I, CTX-II, ADAMTS4, ADAMTS5, RANK, RANKL and TRACP5b mRNA expression in joint fluid of observation group were significantly lower than those of control group whereas OPG and OC contents were significantly higher than those of control group. Conclusion: Arthroscopic debridement combined with intra-articular sodium hyaluronate injection can inhibit the collagen metabolism and bone metabolism disorder mediated by Wnt/β-catenin after surgery.

Clinical effect of Rehabmtation Treating combined with Intra-Articular Injection of Sodium Hyaluronate for Exercise induced knee osteoarthritis

Objective: To study the clinical effect of rehabmtation treating combined with intra-articular injection of sodium hyaluronate for exercise induced knee osteoarthritis, and to research the function, biomechanics of knee joint, and inflammatory factor in articular cavity. Method:Selecting 120 patients with exercise induced knee osteoarthritis, then they were randomly divided into control group (n=60) and intervene group (n=60). The patients in the control group were given conventional rehabmtation treating, and the intervene group were given intra-articular injection of sodium hyaluronate on this basic, they were treated 4 weeks. Evaluating the function of knee joint;conduct isokinetic muscle strength test (determination of the angIllar velocity were 60°/S, 180°/S);test the ratio of muscle peak torque and four muscles centripetal centripetal peak torque and peak torque, work load of femoral knee joints after (simultaneous determination of the angular velocity were 60°/S);detect the inflammatory factor in the joint fluid. Results: After the treatment, clinical effect in intervene group was superior to that in control group,but had no statistical significance;after the treatment, the score of function of knee joint in the two groups were higher than before treatment, and had statistical significance, and the score of function of knee joint in the intervene group were higher than control group, and the difference had statistical significance;Angular velocity of 60°/S and 180°/S the two groups peak torque, work load and the H/Q values were significantly increased, the intervene group were higher than control group, and the difference had statistical significance;inflammatory factor in the two groups were significantly decreased, the intervene group were lower than control group, and the difference had statistical significance.Conclusions: On the basic of rehabmtation treating, the treatment of intra-articular injection of sodium hyaluronate can improve the function of knee joint through improving the knee muscle strength, the level of their participation in the activities of the biomechanics of knee joint, and the inflammatory factor in the joint fluid.

Perceived Efficacy of Vaginal Dryness Relief: A Comparative Clinical Study between <i>Sodium hyaluronate vaginal gel</i>¹<i>vs. Promestriene cream</i>²

In the study, we report the comparative efficacy of the sodium hyaluronate gel vaginal application and promestriene cream which were tested to treat the gynecological symptoms of people who suffered from vaginal dryness. Method: 35 patients applied sodium hyaluronate vaginal gel one time a day for a period of two times a week during three weeks and other 33 patients using promestriene cream applied it every day for a period of three weeks also. Results: No significant difference was observed between vaginal application of sodium hyaluronate gel and promestriene cream, regarding both, as regarding the dryness of the intimate mucosa (p = 0.786), the attribute of moisturizing properties to the intimate mucosa (p = 0.142), the comfort sensation (p = 0.528), and no significant difference was observed regarding the fragrance of the product (p = 0.088). Conclusion: The similar results between vaginal application of both products support the use of sodium hyaluronate vaginal gel (Lubrinat&reg;) in the initial approach of symptoms of vaginal dryness.

The effectiveness of local anesthetic and sodium hyaluronate injection for the posterior disc displacement: A case report

Posterior disc displacement of the temporomandibular joint is rare. We present a unique treatment method and magnetic resonance (MR) images of this condition. The patient was a 43-year-old male with a chief complaint of abnormal occlusion. Regarding the present status, there was no occlusion on the right side. No temporomandibular joint pain, myalgia, or clicking were observed while mouth opening. On MR images, the posterior disc displacement without reduction on the right was observed and it was assumed that occlusal abnormality was due to this condition. We treated him with the following methods. After injection into the superior articular cavity with 2% lidocaine, a sodium hyaluronate preparation was injected followed by intermaxillary traction applied using rubber. The recovery of occlusion was confirmed in follow-up at 5 days after treatment. In the 13th days, the intermaxillary traction was removed. On MR images, the right disc condition was changed to anterior disc displacement with reduction. We consider our treatment methods are effective for this clinical condition.

Meta-analysis of the traditional Chinese medicine fumigation combined with sodium hyaluronate in the treatment of knee osteoarthritis

Objective:To explore the meta-analysis of traditional Chinese medicine fumigation combined with sodium hyaluronate in the treatment of knee osteoarthritis.Methods: The database was originated from CNKI, Wanfang, Chongqing Weipu database, PubMed and so on. The deadline was April 2017. The database was searched with the keywords of'Chinese herbal fumigation','sodium hyaluronate' and'knee osteoarthritis'. Finally, 13 articles about the treatment of Chinese herbal fumigation combined with sodium hyaluronate were obtained. RCT literatures of knee osteoarthritis identified the outcome indicators as the effective rate and recovery rate. Lysholm's score was observed after treatment in both groups, and all the results were input into Review Manager 5.2 for analysis.Result: A total of 13 RCT literatures about the treatment of knee osteoarthritis with Chinese herbal fumigation and sodium hyaluronate were obtained. The control group was treated with sodium hyaluronate or anti-inflammatory drugs, totaling 619 cases. The study group was treated with sodium hyaluronate and Chinese herbal fumigation, totaling 649 cases. The combined effect of forest map OR=1.82, Z=5.00 (P<0.00001), 95% CI. (1.44, 2.30). Compared with sodium hyaluronate or anti-inflammatory drugs, fumigation with traditional Chinese medicine combined with sodium hyaluronate has obvious advantages in curing knee osteoarthritis. The combined effect of forest chart OR=5.55, Z=7.44 (P<0.00001), 95% CI (3.54, 8.73), indicating that compared with sodium hyaluronate or anti-inflammatory drugs, traditional Chinese medicine is used. Fumigation combined with sodium hyaluronate has obvious advantages in the treatment of knee osteoarthritis;forest map combined effect OR=12.11, Z=5.18 (P<0.00001), 95% CI (7.53, 16.68), indicating that compared with sodium hyaluronate or anti-inflammatory drugs, traditional Chinese medicine fumigation combined with sodium hyaluronate in the treatment of knee osteoarthritis can restore knee joint function. Advantages.Conclusion: Chinese herbal fumigation combined with sodium hyaluronate has obvious advantages in curing and effective rate of knee osteoarthritis, and can better restore knee joint function. Due to the lack of literature and low quality, the exact effect still needs to be confirmed by higher quality RCT literature.

Clinical effect of sodium hyaluronate joint cavity filling combined with exercise therapy on patients with post-traumatic knee arthritis

Objective:To research the sodium hyaluronate joint cavity filling combined with exercise therapy on the indicator of pain,motor function,and levels of inflammatory factors in joint fluid in patients with post-traumatic knee arthritis.Method Totally 100 patients(Department of Orthopaedics,The second Affiliated Hospital to Liaoning University of Chinese Medicine,2013.10-2018.8)with post-traumatic knee arthritis were selected,then they were divided into observation group(n=50)and control group(n=50).The control group was given glucosamine sulfate and sodium hyaluronate joint cavity filling,the observation group was given leg press with visual feedback,they were treated 4 weeks.Assessing the pain degree through visual analog scale(VAS),hospital for special surgery knee score(HSS)as the indicator of motor function,inflammatory factor(TNF-α,IL-6,and IL-8)in the joint fluid.Results The score of VAS were lower than pretherapy(P<0.05),the degree of VAS of observation group were lower than control group(P<0.05).The score of HSS were higher than pretherapy(P<0.05),the degree of VAS of observation group were higher than control group(P<0.05).The inflammatory factor(TNF-α,IL-6,and CRP)in the joint fluid were lower than pretherapy(P<0.05),and the observation group were lower than control group(P<0.05).Conclusions Giving sodium hyaluronate joint cavity filling combined with exercise therapy can improve the function of joint.

Study on sodium hyaluronate product as inhibitor of pain due to epidural cicatrization

Objective To evaluate the clinical effect of sodium hyaluronate product(SHP) used as inhibitor to preventing the formation of epidural scar.Method 160 patients suffering from lumbar intervertebral disc herniation were treated by operation,during which SHP was daubed around dura matter as obstruct to prevent epidural scar formation.Result No adverse reaction was observed.Pain was relieved and VAS was decreassed from 8.35 to 0.42 in mean.Using SHP got better curative effect than control group did.The cure and mend rate is 100%.No relapse occurs.Less scar formation was proved by CT in some cases.Conclusion SHP is ideal material used as inhibitor of epidural cicatrization.

火针联合玻璃酸钠注射液关节腔注射对膝骨关节炎患者Lysholm评分和外周血TLR4浓度的影响

目的:观察火针联合玻璃酸钠注射液关节腔注射对膝骨关节炎患者Lysholm评分和TLR4浓度的影响。方法:将80例临床诊断为膝骨关节炎的患者随机分为治疗组和对照组,治疗组采用火针联合玻璃酸钠注射液关节腔内注射治疗,对照组单纯玻璃酸钠注射液关节腔注射,比较治疗前、治疗后5周和12周时Lysholm评分及外周血TLR4浓度的变化。结果:治疗后5周、12周时,治疗组和对照组的膝关节Lysholm评分比较(P=0.000,P=0.000),治疗组和对照组外周血TLR4浓度比较(P=0.000,P=0.000),组间差异具有统计学意义。结论:火针联合玻璃酸钠关节腔注射治疗可提高Lysholm评分、降低血液中TLR4浓度,缓解膝关节疼痛,改善膝关节功能。

SH关节腔内注射治疗兔膝关节炎的实验研究

[目的]探讨透明质酸钠(sod ium hyaluronate,SH)关节腔内注射治疗膝骨性关节炎(OA)的效果。[方法]24只兔子建立骨关节炎动物模型,随机分OA组、SH组、对照组,观察3组软骨、滑膜细胞病理切片及软骨MMP-1免疫组化,进行软骨M ank in s评分,以及检测血液和关节液的IL-1含量的效果。[结果]SH组可见软骨破坏减轻,滑膜纤维增生减少,M ank in s评分有明显改善(P<0.05);血、膝关节滑液IL-1浓度降低(P<0.05),但关节软骨中MMP表达仍然活跃。[结论]SH能减少滑液中炎性介质IL-1的分泌,从而减轻滑膜炎症,缓解对软骨细胞的破坏,延缓OA进展,但仍阻止不了OA进展。

rhBMP-2/SH复合物对微种植体支抗稳定性作用的实验研究

目的评价rhBMP-2/SH复合物对微种植体支抗稳定性的作用。方法实验选用新西兰大白兔12只,每只动物每侧胫骨植入rhBMP-2/SH/微种植体(实验组)、微种植体(对照组)各1枚;术后2、4、8周处死动物进行微种植体周围骨组织形态学观察与微种植体-骨界面剪切力的测量。结果在2、4、8周时,实验组rh-BMP-2/SH/微种植体周围胫骨表面有新骨生成,术后4周和8周时rhBMP-2/SH/微种植体骨质附着量多于单纯微种植体。实验组的剪切力分别为(121.52±20.32)、(213.78±25.30)、(252.15±36.96)N;对照组的剪切力分别为(124.32±18.25)、(175.63±23.45)、(201.07±33.49)N;术后4周和8周时实验组的剪切力大于对照组,比较差异有统计学意义(P<0.05)。结论rhBMP-2/SH复合物可以促进微种植体周围新骨形成,增加骨皮质厚度、骨结合率,从而增加微种植体的稳定性。

丙戊酸钠下调SH-SY5Y细胞中Bcl-2表达的作用及机制分析

目的探讨丙戊酸钠(sodium valproate,VPA)在神经细胞中下调Bcl-2表达的作用及可能机制。方法用不同浓度的VPA处理SH-SY5Y细胞,定量PCR检测Bcl-2 mRNA与miR-34a水平变化,Western blot检测Bcl-2蛋白水平变化,报告基因系统检测Bcl-2启动子活性变化,联合转录抑制剂处理细胞,分析Bcl-2 mRNA稳定性变化,联合miR-34a模拟物及抑制剂处理细胞,进一步分析miR-34a在VPA下调Bcl-2表达中的作用。Annexin V/PI双染结合流式分析细胞凋亡情况。结果 VPA可浓度依赖性下调SH-SY5Y细胞中Bcl-2的mRNA与蛋白水平(P<0.05)。VPA暴露不改变Bcl-2的启动子活性(P>0.05),但显著降低了其mRNA的稳定性(P<0.05)。同时,VPA可上调SH-SY5Y细胞中miR-34a的表达(P<0.05),联合使用miR-34a的抑制剂可逆转VPA对Bcl-2表达的下调作用,且单独的miR-34a模拟物也可以下调细胞中Bcl-2的表达。VPA暴露还可引起SH-SY5Y细胞的凋亡增加(P<0.05)。结论 VPA可通过上调SH-SY5Y细胞中miR-34a水平而抑制Bcl-2的表达。

丙戊酸钠通过激活miR-148a-3p/Mcl-1通路促进SH-SY5Y细胞凋亡

目的探讨抗癫痫药丙戊酸钠(sodium valproate,VPA)对神经细胞的凋亡诱导作用及相关机制。方法将人神经母细胞瘤细胞(SH-SY5Y)用不同剂量VPA处理,流式分析细胞凋亡水平变化,Western blot检测Mcl-1蛋白水平变化,定量PCR分析Mcl-1 mRNA与miR-148a-3p水平变化,报告基因实验检测Mcl-1启动子活性变化。用miR-148a-3p抑制剂联合VPA处理细胞,观察miR-184a-3p在VPA下调Mcl-1表达中的作用及对VPA促SH-SY5Y细胞凋亡效果的影响。结果VPA可促进SH-SY5Y细胞凋亡,并抑制SH-SY5Y细胞中Mcl-1的蛋白和mRNA水平(P<0.05)。VPA处理不影响SH-SY5Y细胞中Mcl-1的启动子活性(P>0.05)。VPA可显著增强SH-SY5Y细胞中miR-148a-3p的表达(P<0.05),使用miR-148a-3p抑制剂可减轻VPA对Mcl-1表达的抑制作用,并减弱VPA的促细胞凋亡效果。结论VPA可能通过激活miR-148a-3p/Mcl-1信号通路促进SH-SY5Y细胞凋亡。