Treatment Of Shaoyaogancao Decoction On Pancreatic Type Of Oddi Dysfunction And The Study Of L-type Calcium Channel Activity Regulation Mechanism

Background: In1887Italy physiologist Ruggero Oddi described thedescending portion of duodenum biliary and pancreatic duct openings exist sphincter forthe first time, which was named the sphincter of Oddi (Sphincter of Oddi, SO) after that.Oddi Sphincter (Sphincter of Oddi, SO) is independent of the duodenal wall, openended in duodenal papilla Sphincter, is the only portal bile discharge biliary, regulatingthe secretion of bile pancreatic juice, maintaining the balance of low pressure, low flowinside the bile duct[2,3].According to the SOD classification of Rome III criteria, SOD isdivided into the pancreatic duct of SO dysfunction and SO dysfunction of bile duct.Duct type SOD can be divided into Ⅰ,Ⅱ,Ⅲ. The present study was performed toinvestigate the alleviating effects of Shaoyaogancao Decoction on Pancreatic TypeSOD.Peony liquorice decoction originated in Zhang Zhongjing article29in treatise onfebrile disease. Peony liquorice decoction is composed of white peony, licorice[4], andexerts antispasmodic effects duo to the active ingredient of paeoniflorin., in the Theactive ingredient of radix glycyrrhizae is glycyrrhizic acid, which has anti-inflammatoryeffect. Research showed that peony liquorice decoction is used as Pain Medication andexerts sedative and analgesic and smooth muscle relaxation effects, currently in clinic, Peony liquorice decoction has been widely used in respiratory and digestive systemsymptoms in clinically[5].Analgesic effect peony liquorice decoction priorities related toAnti-spasticity effects of Peony liquorice decoction is involved in its effects on relievepain.Research showed that hypercholesterolemia in rabbit leads to motility disorder anddysfunction of SOD.[7].L type calcium channel is the voltage-gated calcium whichdetermines smooth muscle contractile function, the alpha1subunit is the functional one.It has been confirmed that calcium triggers muscle contraction through the influx of L-type calcium channel. Literature shows paeoniflorin Peony (Paeoniflorin, PF) can relaxgastrointestinal smooth muscle and isolated rabbit SO muscle. However, the underlyingmechanism of the relaxation of paeoniflorin remains largely unclear.To further explorethe underlying mechanism involved in the relaxation of paeoniflorin, the present studywas performed to investigate the effects of PF on gene and protein expression ofsubunit in hypercholesterolemia rabbit cells. Whole cell patch clamp was also used toinvestigate the effects of different concentration of PF on L-type calcium current (ICa,L).Objectives:Part1Peony liquorice decoction treatment in patients with pancreatic SOD.Part21. Gene and protein expression of L type calcium channel alpha1c subunit inhypercholesterolemia rabbits;2. Effects of different concentrations of PF on alpha1c subunit mRNA and proteinexpression in hypercholesterolemia rabbits;3. Effects of PF on L-type calcium current in hypercholesterolemia rabbits andhealthy ones;4. Mechanism involved in relaxation effects of PF.Methods:Part1: 1. Acquisition of dalian medical university first affiliated hospital outpatient andinpatient specimens, patients in the outpatient clinic or hospital for January2009-December2010, collected patients after cholecystectomy, conform to the RomeⅢcriteria for the diagnosis and conform to the pancreatic SOD diagnose patients into thegroup, the group of cases, a total of39(including2cases of type Ⅰand type Ⅱ13cases,24cases of type Ⅲ), after the group were selected patients were randomlydivided into2groups:1.1The treatment group,23cases,10were male, female13cases, aged35to58years old, average age59.3+/-18.8;First onset time of postoperative16~57months,an average of20.3+/-2.4months (type I in0cases of pancreatic type, type Ⅱ in5cases, Ⅲ10);1.2The control group:16cases,5cases of male, female10, cases were aged35to58, an average of52.7+/-20.1years old;First onset time of16~57monthspostoperatively, an average of19.4+/-2.9months (including biliary type Ⅰ in0cases,1cases type Ⅱ, Ⅲ1cases).2. Two groups of patients admitted to hospital after correct electrolyte disorders,such as anti-infection treatment, did not give spasmolysis drug in the treatmentprocess.Control group oral placebo (herbal tea), the treatment group oral peonyliquorice decoction.3. Peony liquorice decoction of traditional Chinese medicine treatment effect wasobserved, and in a period of12months follow-up, patients with pain relief, pancreaticfunction recovery as observation indexes, the curative effect evaluation.Part2:1. Health pure New Zealand white rabbit were randomly divided into two groups,the control group (Concrol, CON) and high cholesterol group (Hypercholesterolemia,HC). Control group were feeded with standard food, hypercholesterolemia group werefeeded with standard and cholesterol food for8weekds to induce hypercholesterolemia.2. SO cells in CON group and HC group were extracted, primitive culture, immunehistochemical methods were made. SO cells in HC group were divided into three groups, incubated with medium containing10-7mol/L,10-6mol/L,10-5mol/L PF for12h. Ltype calcium channel alpha1c subunit mRNA and protein expression in each groupwere performed using Real-time PCR and Western Blotting asays..3. Whole cell patch clamp was used to investigate the effects of differentconcentration of PF on ICa, L.Results:Part1:Effective16cases in treatment group (type Ⅰ,0case, type Ⅱ3cases effective,Ⅲ valid13cases), the total effective rate (69.6%).The control group recurrence rate12.5%;Effective2cases in (typeⅠ0cases, type Ⅱ1case,1case of type Ⅲ),1years recurrence4cases in treatment group (type Ⅱ in1cases,3type Ⅲ).The rateof25%.1years recurrence2cases in control group (type Ⅱin1cases,1typeⅢ).The rate of100%.Part2:1. L type calcium channel alpha1c subunit mRNA and protein expression levelwas increased in hypercholesterolemia rabbits;2. PF exerted significant inhibitory effects on mRNA and protein expression of l-typecalcium channel alpha1c subunit, and the effects were concentration-dependent;3. Whole cell patch clamp assay showed that PF, in the dose of0.1μmol/L1μmol/L,10μmol/L， exerted significant inhibitory effects on ICa, shifted LV curve, andthe effects were concentration-dependent.Conclusions:Part1Reliving effects of Oral peony liquorice decoction on pancreatic type Ⅱand typeⅢ SOD after cystic resection treatment effect is remarkable, the reliving effects need ashort course, act quickly, improve liver function and and with a lower recurrence rateafter treatment, peony liquorice decoction exerts no effects on type I SOD.Part21. Change in alpha1c preotin expression is one of the mechanism involved in the rise of calcium activity in rabbits. PF relaxs SO by decreasing calcium activity;2. Inhibition of PF on calcium channel alpha1c subunit mRNA and protein expressionlevel role was dose-dependent, and PF in10μmol/L exerted stronger effects;3. PF exerted significant inhibition effects on ICa, L，which could be one of themechanisms involved in relaxing smooth muscle.