Study On Antiviral,Anti-inflammatory,Immunomodulatory And Antipyretic Pharmacological Effects Of Marine Traditional Chinese Medicine Ipomoea Pes-caprae (Linn.) Sweet

Objective:To evaluate the antiviral,anti-inflammatory,immunomo-dulatory and antipyretic pharmacological activities of extracts from leaves of Ipomoea pes-caprae（Linn.）Sweet,in order to develop new applications and provide references for clinical medication.Methods:（1）Use the method of the cold immersion to obtain the ethanol extract of Ipomoea pes-caprae（Linn.）Sweet（IPEE）;use the method of two-phase extraction to prepare the petroleum ether fraction（IPPE）,ethyl acetate fraction（IPEAE）,n-butanol fraction（IPNE）;use the method of HPLC to separate IPEE and separate IPPE by TLC.（2）Cytotoxicity to IPEE fraction and IPPE fraction on Huh-7,JEG-3 and Hela cells by Alamar blue method.The activity of IPEE fraction and IPPE fraction on dengue virus（DENV）and Zika virus（ZIKV）Chikungunya virus（CHIKV）virus titer was detected by plaque assay.（3）Establish LPS-induced RAW264.7 cell inflammation model,detect NO,TNF-α,IL-6,and IL-1βcontent in cell supernatant,and detect anti-inflammatory activity of IPEE,IPPE,IPEAE and IPNE.（4）Establish a mouse cyclophosphamide-induced immunesuppressive model,determine the immune organ coefficient,NK cell proliferation rate,T lymphocyte and B lymphocyte proliferation rate,serum IL-2,TNF-α,IFN-γcontent The spleen tissue ACP and LDH activities were evaluated to evaluate the immunomodulatory activity of IPEE.（5）To establish a model of rat dry yeast heat,and to determine the body temperature change,maximum body temperature rise,△T_max,serum IL-1β,TNF-αand PGE2 levels before and after modeling,so as to evaluate the antipyretic activity of IPEE.Results:（1）A total of 5 IPEE fractions and 6 IPPE fractions were prepared.（2）The fractions with good anti-DENV activity in all fractions were IPEEF5,IPPEF2 and IPPEF3.Among the three fractions,IPPEF2 was better for anti-ZIKV activity,and IPPEF2 and IPPEF3 were better for anti-CHIKV activity.It indicated that IPEE has good antiviral effect.（3）LPS induced RAW264.7 cell inflammation model.The best modeling conditions were 1μg/m L LPS induction for 24h.IPEE,IPPE and IPEAE significantly decreased LPS-induced RAW264.7 cells.The release of NO increased,with the lowest IPPE inhibition concentration(IC₅₀),which was13.88μg/m L.IPEE,IPPE,IPEAE and IPNE all significantly reduced the increase of TNF-α,IL-6 and IL-1βsecretion in RAW264.7 cells induced by LPS.IPEE and IPPE had better inhibitory effect.It shows that IPEE shows better anti-inflammatory activity in the in vitro inflammation model.（4）The high-dose IPEE group significantly inhibited the reduction of the spleen coefficient of immunosuppressed mice induced by cyclophosphamide（P<0.01）,but did not significantly inhibit the decrease of thymus coefficient（P>0.05）;IPEE high and medium dose groups could The activity of NK cells in immunosuppressed mice was significantly decreased（P<0.05,P<0.05）.The high and medium doses of IPEE significantly increased the proliferation rate of T lymphocytes after 48 hours of induction（P<0.01,P<0.01）.The high,medium and low dose groups of IPEE significantly increased the T lymphocyte proliferation rate after 72 hours of induction（P<0.01,P<0.01）.The IPEE high and middle dose groups significantly increased B lymphocytes after 48 hours of induction.The cell proliferation rate（P<0.01,P<0.01）,IPEE high dose group can significantly increase the B lymphocyte proliferation rate after 72h induction in vitro（P<0.01）;IPEE high dose group can significantly inhibit mouse serum IL-2The levels of TNF-αand IFN-γwere decreased（P<0.01,P<0.01,P<0.05）.It showed that IPEE had a significant effect on improving immunity（5）IPEE did not significantly inhibit the increase of body temperature in dry yeast-induced rats,but the high-dose group significantly inhibited the increase of TNF-αand IL-1βlevels（P<0.01,P<0.05）.Conclusion:This study conducted antiviral,anti-inflammatory,immunomodu-latory and antipyretic pharmacological activities on the leaves of Ipomoea pes-caprae（Linn.）Sweet.The results showed that the thick vine leaves had better anti-dengue virus,Zika virus and chikungunya virus activity.It has better anti-inflammatory and in vivo immunomodulatory activities,but has no significant antipyretic effect.The study on the pharmacological activity of the Ipomoea pes-caprae（Linn.）Sweet increases the applicable range of the thick vine,provides a theoretical basis for improving the application value of Ipomoea pes-caprae（Linn.）Sweet,and also provides a reference for guiding the drug.