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BackgroundHerbal　Pyrola　is　the　whole　plant　of　Pyrola　Calliantha　H.　Andres,　Pyrola　decorrata　H.　Andres　or　others　at　the　same　category.　According　to　Chinese　Pharmacopoeia　(edition　in　2000),　it　is　attributed　to　the　channels　of　liver　and　kidney　and　has　the　effects　of　eliminating　wind　and　damp,　strengthening　bones　and　muscles,　adjusting　blood　and　essence,　and　reinforcing　insufficiency　and　kidney.　In　abroad,　the　chemical　constituents　of　Herbal　Pyrola　are　limitedly　studied　on　P.japonica　Klenze　ex　Alef,　P.incarnata　H.　Andres　and　P.renifolia　H.　Andres.　In　recent　years,　intensive　studies　about　it　have　being　performed　in　China.　In　light　of　these　studies,　Herbal　Pyrola　is　rich　in　flavonoids,　tannin,　benzoquinones,　polyphenols　and　others.　Flavonoids　are　major　elements　and　have　many　pharmacological　effects.　2’’-o-galloyhyperin　belongs　both　to　tannin　and　to　flavonoids　because　it　has　2-phenylchromone.　It　is　held　in　Traditional　Chinese　Medicine　that　combination　of　Herbal　Pyrola　with　other　herbs　is　able　to　treat　some　diseases,　such　as　deficient　type　asthma,　yang　deficiency　of　kidney,　low　back　pain,　asthenia　in　feet　and　knees,　hemiplegia,　intestinal　catarrh,　abscess,　and　metrorrhagia,　etc.　Pharmacological　studies　suggest　that　Herbal　Pyrola　has　many　effects,　such　as　strengthening　cardiac　muscle,　decreasing　blood　pressure,　anti-inflammation,　alleviating　symptoms　of　coronary　heart　disease　and　fighting　against　aging.　It　is　reported　that　Herbal　Pyrola　has　ability　to　dilate　blood　vessel　of　limbs,　ears　and　brains　in　vitro　and　to　increase　cardiac　nutritive　blood　in　vivo.　Herbal　Pyrola　is　also　capable　of　elevating　cAMP　in　plasma,　eliminating　oxygen-derived　free　radicals　and　lowering　cerebral　vessel　resistance.　Moreover,　Herbal　Pyrola　had　been　clinically　used　to　treat　coronary　heart　disease,　hypertension,　angitis　and　pulmonary　tuberculosis　etc.AimThis　study　aimed　at　exploring　the　effective　and　economical　methods　of　extracting　Total　Flavonoids　from　Herbal　Pyrola　(TFHP)　and　studying　pharmacological　effects　on　acute　myocardial　ischemia　and　acute　toxicity　to　accumulate　the　evidences　and　bases　for　developing　a　new　drug　from　TFHP.Methods1．The　extract　procedure　of　TFHPThe　whole　plant　of　Herbal　Pyrola　broken　up　was　boiled　and　regurgitated　with　70%　alcohol　for　three　times（2.0h,　1.5h　and　1.0h　respectively）under　the　normal　pressure,　and　then　condensed　under　the　negative　pressure　to　obtain　condensing　liquid　1　and　condensing　sediment　1.　Condensing　sediment　1　was　extracted　with　boiling　water　for　2.0h　under　the　normal　pressure　and　the　extracted　liquid　was　filtered,　condensed　and　evaporated　under　the　negative　pressure　to　obtain　condensing　liquid　2　and　condensing　sediment　2.　Then,　condensing　sediment　2　was　discarded.　Condensing　liquid　1　and　condensing　liquid　2　were　mixed　up　to　get　condensing　liquid　3.　After　condensing　liquid　3　was　solved　with　20%　alcohol　and　filled　it　in　the　column　of　macroporous　adsorbing　resins,　the　column　of　macroporous　adsorbing　resins　was　successively　flushed　with　the　different　concentrations　of　alcohol　(20%,　50%　and　90%　respectively).　As　a　consequence,　the　extract　that　was　separated　with　50%　alcohol　was　obtained　by　evaporation　of　solvent.　With　the　methods　of　ultraviolet　spectrophotography　and　aluminum　trichloride　color,　amount　of　the　extract　was　measured　on　the　basis　of　2’’-o-galloyhyperin　as　standard　component.　2．The　acute　toxicity　of　TFHPTen　ICR　mice　were　used　in　the　preliminary　experiment　to　look　at　general　toxicity　and　death　of　mice　after　intragastric　administration　of　TFHP　in　maximal　concentration　(20%)　and　0.2mL/10g　or　0.4mL/10g.　The　toxic　reactions　were　observed　for　7　days.　On　the　basis　of　the　preliminary　result,　20　ICR　mice　were　used　to　measure　the　maximal　administration　dose　(MAD)　in　the　normal　experiment.　20%　TFHP　(0.4mL/10g)　was　intragastrically　gived　to　mice　two　times　in　the　first　day.　Then,　mice　were　observed　for　7days.3．The　protective　effect　of　TFHP　on　acute　myocardial　ischemia　induced　by　Pit　in　rats　Sixty　male　SD　rats　were　randomized　into　control　group,　model　group,　positive　control　group　and　TFHP　groups　(30,　15　and　7.5mg/kg　respectively).　Compound　salvia　drop　pill　(CSDP,　290mg/kg)　was　given　to　rats　in　positive　control　group,　TFHP　to　rats　in　the　drug-treated　group　and　distilled　water　to　rats　in　control　group　and　model　group　for　7　days　by　intragastric　administration.　The　acute　myocardium　ischemia　was　induced　by　sublingual　venous　injection　of　Pit　(0.　5U/kg)　after　1h　of　the　last　administration.　The　changes　of　the　T　wave　height　on　II　channel　electrocardiograph　(ECG)　and　the　latent　period,　lasting　time,　times　and　rate　of　arrhythmias　were　observed　before　and　after　Pit　injection.4．The　protective　effect　of　TFHP　on　AMI　induced　by　the　ligation　of　left　descending　coronary　artery　in　ratsSixty　male　SD　rats　were　randomized　into　sham　group,　model　group,　positive　control　group,　and　TFHP　groups　(240,　120　and　60mg/kg　respectively).　Nifedipine　(2mg/kg)　was　given　to　rats　in　positive　control　group,　TFHP　to　rats　in　the　drug-treated　group　and　distilled　water　to　rats　in　control　group　and　model　group　for　7　days　by　intragastric　administration.　The　model　of　acute　myocardial　infarction　(AMI)　was　established　by　ligating　the　left　descending　coronary　artery　with　a　ligature　(5-0　silk　suture).　The　changes　of　T　wave　height　(II　channel　ECG)　were　observed　before　and　after　ligating　the　left　descending　coronary　artery.　After　myocardial　infarction　for　24h,　the　changes　of　the　T　wave　height,　size　of　myocardial　infarction,　activities　of　creatine　phosphokinase　(CK),　lactate　dehydrogenase　(LDH)　and　superoxide　dismutase　(SOD)　and　levels　of　nitric　oxide　(NO),　free　fat　acid　(FFA)　and　malondialdehyde　(MDA)　in　serum　were　determined.Results1．The　extraction　and　identification　of　TFHPThe　results　showed　that　the　extract　content　of　TFHP　was　20.86%　and　the　extract　rate　was　8.89%.2．The　acute　toxicity　of　TFHPMice　did　not　show　any　abnormality　and　death　during　the　preliminary　experiment　and　it　was　suggested　that　the　LD50　of　TFHP　was　not　detected.　Instead,　MAD　was　examined.　The　result　indicated　that　MAD　of　TFHP　was　16g/kg.3．The　protective　effect　of　TFHP　on　acute　myocardial　ischemia　induced　by　Pit　inratsThe　results　displayed　that　the　height　of　T　waves　was　raised,　the　latent　period　of　arrhythmias　was　prolonged　and　the　lasting　time,　times　and　rate　of　arrhythmias　were　decreased　after　TFHP　pretreatment　in　comparison　with　the　model　group　(P＜0.05　or　0.01).4．The　protective　effect　of　TFHP　on　AMI　induced　by　the　ligation　of　left　descending　coronary　artery　in　ratsThe　results　showed　that　pretreatment　with　TFHP　was　capable　of　elevating　the　height　of　T　waves,　reduced　the　size　of　myocardial　infarction,　decreased　the　activities　of　CK　and　LDH　and　the　levels　of　FFA　and　MDA　,　and　increased　the　release　of　NO　and　the　activity　of　SOD　in　comparison　with　the　model　group　(P　＜0.05　or　0.01).ConclusionThe　extract　content　of　TFHP　was　20.86%　and　the　extract　rate　of　TFHP　8.89%.　MAD　of　TFHP　was　16g/kg.　TFHP　had　an　ability　to　protect　myocardium　from　ischemic　injury　induced　by　Pit　and　the　ligation　of　left　descending　coronary　artery　possibly　via　antioxidation　and　modulation　of　metabolism　disorders.

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