ประสิทธิภาพของสารสกัดพอลิแซ็กคาไรด์จาก Arthrospira platensis และสาหร่ายสีเขียวขนาดใหญ่บางชนิดในการยับยั้งเชื้อไวรัสก่อโรคเริมและเซลล์มะเร็งปากมดลูก

Abstract

Herpes simplex infection is caused by herpes simplex virus (HSV). The infection remains a prominent problem on human health because the viruses can cause a latent infection. The purpose of this study was to investigate the inhibitory effect of the polysaccharide extract from Arthrospira platensis, Cladophora spp., Spirogyra spp. and Ulva reticulata against herpes simplex virus and cervical cancer cell. The polysaccharides extract contain sugar (47.44 - 57.79 %), protein (13.69 - 22.80 %) and sulfate (0.30 - 7.31 %). The chemical characteristic of polysaccharide extract was analyzed by FT-IR technique. The polysaccharide extract represents C – H and C – O – C group of glycosidic linkage of polysaccharides, S = O and C – O – S group of sulfate groups. The toxicity of polysaccharide extract on Vero cell was determined by MTT assay. The polysaccharide extract from A. platensis, Cladophora spp. and U. reticulata showed low toxicity on the cell with CC50 was greater than 5,000 μg/mL, whereas the polysaccharide extract from Spirogyra sp. showed CC50 of 778.31 μg/mL. Then, antiviral activity of the polysaccharide extract against HSV was evaluated by plaque reduction assays. The results indicated that polysaccharide extracts from A. platensis, Cladophora spp. and U. reticulata showed antiviral activity against HSV-1 upon treatment before viral attachment with EC50 of 5,748.10, 70.31 and 2,525.90 μg/mL, respectively. The antiviral activity against HSV-2 upon treatment before viral attachment with EC50 of 286.11, 5.85 and 163.26 μg/mL, respectively. Additionally, the polysaccharide extracts from A. platensis, Cladophora spp., Spirogyra spp. and U. reticulata showed antiviral activity against HSV-1 upon treatment during viral attachment with EC50 of 1,282.90, 15.17, 40.66 และ 58.32 μg/mL, respectively. The antiviral activity against HSV-2 upon treatment during viral attachment with EC50 of 126.46, 2.57, 22.49 and 9.70 μg/mL, respectively. Furthermore, the polysaccharide extracts from A. platensis, Cladophora spp. and U. reticulata showed antiviral activity against HSV-1 upon treatment after viral attachment with EC50 of 1,186.40, 9.78 and 263.95 μg/mL, respectively. The antiviral activity against HSV-2 upon treatment after viral attachment with EC50 of 2,203.50, 26.96 and 527.28 μg/mL, respectively. The inactivation of virus particle was examined by virucidal assay and the inhibition of viral multiplication was determined by viral growth kinetics assay. The polysaccharide extract was demonstrated the highest direct inactivation of HSV-1 virions at 4 hours after treatment by viral load reduced of 0.42 - 3.65 log PFU/mL. The highest direct inactivation of HSV-2 virions at 4 hours after treatment by viral load reduced of 2.11 - 3.01 log PFU/mL. Moreover, the inhibitory effect on HSV-1 multiplication was observed to be the highest at 36 hours after treatment with polysaccharide extract by viral load reduced of 0.61 - 7.51 log PFU/mL. The multiplication of HSV-2 was also showed the highest inhibition after treatment with the algal polysaccharide extracts for 36 hours by viral load reduced of 0.35 - 5.38 log PFU/mL. The inhibition of cervical cancer cell proliferation was evaluated by MTT assay. The polysaccharide extract from Spirogyra spp. inhibited the cervical cancer cell proliferation with EC50 of 1,522.60 μg/mL at 72 hours after treatment. Besides, the polysaccharide extract could induce apoptosis on the cervical cancer cell. On the contrary, the polysaccharide extracts of A. platensis, Cladophora spp. and U. reticulata at the concentration of 10,000 μg/mL inhibited the cervical cancer cell proliferation less than 50%. The polysaccharide extract from A. platensis, Cladophora spp. and Spirogyra spp. presented the inhibition of cervical cancer cell colony formation by clonogenic assay with EC50 of 745.16, 843.10 and 2,067.80 μg/mL, respectively. Moreover, the polysaccharide extracts from A. platensis, Cladophora spp. and U. reticulata were assessed the inhibition of cervical cancer cell migration by scratch assay with the inhibition of 88.85, 89.45 and 54.51 %, respectively. Therefore, the polysaccharide extract was developed to the soft gelatin capsule product as a potential anti-HSV agent for treatment of HSV infection and cervical cancer. The soft gelatin capsule product was stable at 25 °C for 1 month. Furthermore, the efficiency of product on antiviral activity demonstrated stability at a temperature between 4 - 45 °C for 3 months.