| Background: Cardiovascular disease is an important factor causing human death,and incidence rate and mortality rate are increasing year by year.Cardiovascular diseases mainly include myocardial ischemia or infarction,heart failure and other diseases,mainly due to vascular wall sclerosis leading to progressive stenosis of vascular lumen,which mainly due to vascular wall sclerosis leading to progressive stenosis of vascular lumen,resulting in ischemia and hypoxia.Meanwhile,due to the overload of calcium and massive reactive oxygen species production,which is accompanied by the development of inflammatory reaction.LPS can induce myocarditis,which is highly correlated with cardiovascular disease.Therefore,reducing inflammatory response may be a potential measure or target for the treatment of cardiovascular disease.Icariin is the main active component of the traditional Chinese medicine Epimedium.It has anti-inflammatory and cardiovascular protection effects,and has unique pharmacological effects on inflammatory cardiovascular diseases.However,its hydrophobicity and lower bioavailability limit its clinical application.Objective: This study is to design a new type of nano-drug mPEG-ICA(d)NPs to change the hydrophobicity of icariin,increase drug loading and drug release duration of action,improve its drug delivery,enhance its targeting,which provide an experimental basis for the clinical application of icariin.Methods:First,hydrophilic polyethylene glycol monomethyl ether(m PEG)was grafted with succinic anhydride(SA)to form a polyethylene glycol-icariin(mPEG-ICA)polymer.mPEG-ICA(d)nanoparticles were prepared by physical embedding of ICA and dialysis method.The structure of the mPEG-ICA polymer was characterized by Fourier Infrared Spectroscopy and Nuclear magnetic resonance spectroscopy.Then,to measure mPEG-ICA(d)nanoparticles’particle size and zeta potential,observe its morphology,and examined its drug loading and encapsulation efficiency as well as its release degree in solutions with different p H values,optimize the prepareation parameter,and test the cytotoxicity in vitro.Using LPS induced inflamation model to verify the anti-inflammation functionin of mPEG-ICA(d)NPs.the MTT experiment was used to detect the viability of H9c2 cells.Lactate dehydrogenase(LDH)release was observed to determine cell damage,Cell apoptotis was detected by Hoechst 33258 stainaing,TUNEL and Annexin V-FITC/PI staining;Using RT-q PCR methods were to detect effect of the NPs on the mRNA expression of inflammation markers TNF-α,IL-1βand IL-6 in H9c2cells induced by LPS.Results:1.The nanoparticle mPEG-ICA(d)was successfully prepared.the nanoparticles have regular morphology,spherical shape,uniform size.The average partical size of(220.0±13.7)nm,the zeta potential of(2.30 ± 1.33)m V.The content of ICA in nanoparticles measured by Ultraviolet Spectrophotometry was(21.9±0.013)%,and the encapsulation efficiency was(54.9±0.032)%,it can be prepared repeatedly and have good biological safety;2.In the p H7.4 PBS solution,the release of free ICA drug reached to(77.21 ±0.15)% within 12 hours,while the mPEG-ICA(d)NPs reached to(52.80 ± 1.70)%within 72 hours.In the p H6.8 PBS solution,the cumulative drug release of mPEG-ICA(d)NPs within 48 h was reached to(75.66±0.17)%.this showed that nanoparticle was more easily release ICA in the acidic environment;3.mPEG-ICA(d)NPs can reduce cell damage induced by LPS,such as LDH and cell apoptosis rate decreasing.what’s more,inflamatory markers TNF-α,IL-1β and IL-6mRNA were inhibited by mPEG-ICA(d)NPs.Conclusion:1.A new type of mPEG-ICA(d)NPs was successfully prepared;2.In mPEG-ICA(d)NPs,the ICA release in acidic mediators is higher than that in neutral solutions,the release characteristics of mPEG-ICA(d)NPs may be beneficial to the local treatment of myocardial inflammatory injury;3.mPEG-ICA(d)NPs can reduce LPS-induced cell damage,and inhibit the mRNA expression of pro-inflammatory cytokines,the results indicated that mPEG-ICA(d)NPs may have a protective effect on LPS induced myocarditis. |
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